@article {pmid1647500, year = {1991}, author = {Garofalo, O and Kennedy, PG and Swash, M and Martin, JE and Luthert, P and Anderton, BH and Leigh, PN}, title = {Ubiquitin and heat shock protein expression in amyotrophic lateral sclerosis.}, journal = {Neuropathology and applied neurobiology}, volume = {17}, number = {1}, pages = {39-45}, doi = {10.1111/j.1365-2990.1991.tb00692.x}, pmid = {1647500}, issn = {0305-1846}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aged ; Amyotrophic Lateral Sclerosis/immunology/*metabolism/pathology ; Antibodies, Monoclonal ; Brain/pathology ; Female ; Heat-Shock Proteins/*biosynthesis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Spinal Cord/metabolism/pathology ; Ubiquitins/*biosynthesis ; }, abstract = {The expression of two heat shock proteins, HSP72 and p57, in addition to ubiquitin, has been studied immunocytochemically in nine amyotrophic lateral sclerosis (ALS) cases and 10 age-matched controls. HSP72 and p57 antibodies did not identify the characteristic ubiquitin-immunoreactive inclusions present in anterior horn cells in ALS spinal cord. Antibodies to HSP72, but not to p57 or ubiquitin, strongly labelled structures corresponding to polyglucosan bodies in spinal grey matter. Such immunoreactive profiles were more abundant in ALS cases, although they were also present in control material. They were sometimes identified by haematoxylin and eosin and periodic acid Schiff reaction, but were not labeled by phosphotungstic acid haematoxylin or by antibodies to glial fibrillary acidic protein. Although ubiquitin, HSP72 and p57 are stress-induced proteins, they are expressed differently and might therefore have different significance in neuronal degeneration.}, }
@article {pmid1750186, year = {1991}, author = {Martin, JE and Sosa-Melgarejo, JA and Swash, M and Mather, K and Leigh, PN and Berry, CL}, title = {Purkinje cell toxicity of beta-aminopropionitrile in the rat.}, journal = {Virchows Archiv. A, Pathological anatomy and histopathology}, volume = {419}, number = {5}, pages = {403-408}, pmid = {1750186}, issn = {0174-7398}, mesh = {Aminopropionitrile/*adverse effects ; Animals ; Female ; Immunohistochemistry ; Injections, Intraperitoneal ; Kyphosis/chemically induced/diagnostic imaging ; Purkinje Cells/*drug effects/metabolism/pathology ; Radiography ; Rats ; Rats, Inbred Strains ; Scoliosis/chemically induced/diagnostic imaging ; }, abstract = {Compounds causing neurolathyrism are putative aetiological agents in neurodegenerative disorders including amyotrophic lateral sclerosis. beta-Aminopropionitrile (BAPN) is one such compound. We have administered this lathyrogenic agent at a dose of 1 g/kg by the intraperitoneal route in experiments in adult Sprague-Dawley rats during a period of 10 weeks. The rats developed marked kyphoscoliosis, ataxia with paralysis and muscle wasting of the hind limbs. Vacuolation and loss of Purkinje cells developed, but no anterior horn cell degeneration was noted. Immunohistochemical studies of phosphorylated neurofilaments and the 72 kDa heat shock protein were normal and no intraneuronal ubiquitinated inclusions were seen. High-dose intraperitoneal BAPN in the rat causes Purkinje cell changes, but no other central nervous system abnormalities.}, }
@article {pmid2054224, year = {1991}, author = {Namba, Y and Tomonaga, M and Ohtsuka, K and Oda, M and Ikeda, K}, title = {[HSP 70 is associated with abnormal cytoplasmic inclusions characteristic of neurodegenerative diseases].}, journal = {No to shinkei = Brain and nerve}, volume = {43}, number = {1}, pages = {57-60}, pmid = {2054224}, issn = {0006-8969}, mesh = {Alzheimer Disease/metabolism ; Cytoplasmic Granules/*metabolism ; Dementia/metabolism ; Heat-Shock Proteins/*metabolism ; Humans ; Immunohistochemistry ; *Nerve Degeneration ; Nervous System Diseases/*metabolism/physiopathology ; Parkinson Disease/metabolism ; }, abstract = {Several degenerative diseases of the central nervous system are characterized by the presence of neuronal inclusions. One of these inclusions, neurofibrillary tangles in Alzheimer's disease, has been shown to contain ubiquitin that belongs to a group of proteins known as heat shock proteins. Subsequent studies revealed that ubiquitin is also associated with various neuronal inclusions including Lewy bodies, Pick bodies and hyaline inclusions. Very recently, ubiquitin has been found also to be associated with glial inclusions that are unique to multiple system atrophy. The close association of ubiquitin with varying cellular inclusions, together with its function in the proteolytic process, raised the hypothesis that ubiquitin may be involved in the degradation of abnormal proteins appearing in the damaged neurons and glial cells. In central nervous system, a group of heat shock proteins collectively known as HSP 70 is also present which is constitutive and/or inducible. Since HSP 70 has been suspected to play a crucial role degradation and repair of abnormal intracellular proteins, we hypothesized that HSP 70 may be associated with those inclusions, as the case with ubiquitin. To test this we performed immunohistochemical studies on brain tissues from patients with various neurodegenerative conditions by using specific polyclonal antibody to HSP 70. Brain tissues were obtained at autopsy from each three patients with Alzheimer's disease, Pick's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and multiple system atrophy. Tissues were fixed in buffered formalin and embedded in paraffin. Immunostaining was performed by the standard ABC method using diaminobenzidine as a chromogen. Sections were lightly stained with hematoxylin. Polyclonal antibodies were raised in rabbits against mouse HSP 70.(ABSTRACT TRUNCATED AT 250 WORDS)}, }
@article {pmid7810588, year = {1994}, author = {Goodman, AB}, title = {Elevated risks for amyotrophic lateral sclerosis and blood disorders in Ashkenazi schizophrenic pedigrees suggest new candidate genes in schizophrenia.}, journal = {American journal of medical genetics}, volume = {54}, number = {3}, pages = {271-278}, doi = {10.1002/ajmg.1320540317}, pmid = {7810588}, issn = {0148-7299}, mesh = {Age Factors ; Amyotrophic Lateral Sclerosis/ethnology/*genetics ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Chromosomes, Human, Pair 21 ; Female ; Hematologic Diseases/ethnology/*genetics ; Humans ; Jews ; Male ; Pedigree ; Risk Factors ; Schizophrenia/ethnology/*genetics ; }, abstract = {Among relatives of Ashkenazi schizophrenic probands the rate of amyotrophic lateral sclerosis was 3/1,000, compared to expected population rates of approximately 2/100,000. Relative risk of bleeding disorders, including hematologic cancers, was increased more than three-fold compared to controls. Co-occurrence of motor neuron disease and blood dyscrasias, accompanied by psychosis, has long been recognized. A virally mediated autoimmune pathogenesis has been proposed. However, the familial co-occurrence of these three disease entities raises the possibility that the disease constellation be considered as a manifestation of a common underlying genetic defect. Such expansion of the spectrum of affectation might enhance the power of both candidate gene and linkage studies. Based on these findings the loci suggested as candidate regions in schizophrenia include a potential hot spot on chromosome 21q21-q22, involving the superoxide dismutase and amyloid precursor protein genes. Alternatively, genes on other chromosomes involved in the expression, transcription, or regulation of these genes, or associated with the illnesses of high frequency in these pedigrees are suggested. Candidates include the choroid plexus transport protein, transthyretin at 18q11.2-q12.1; the t(14;18)(q22;21) characterizing B-cell lymphoma-2, the most common form of hematologic cancer; and the 14q24 locus of early onset Alzheimer's disease, c-Fos, transforming growth factor beta 3, and heat shock protein A2. Expression of hematologic cancers and the suggested candidate genes are known to involve retinoid pathways, and retinoid disregulation has been proposed as a cause of schizophrenia.}, }
@article {pmid8190301, year = {1994}, author = {Gao, YL and Raine, CS and Brosnan, CF}, title = {Humoral response to hsp 65 in multiple sclerosis and other neurologic conditions.}, journal = {Neurology}, volume = {44}, number = {5}, pages = {941-946}, doi = {10.1212/wnl.44.5.941}, pmid = {8190301}, issn = {0028-3878}, support = {NS 07098/NS/NINDS NIH HHS/United States ; NS 08952/NS/NINDS NIH HHS/United States ; NS 11920/NS/NINDS NIH HHS/United States ; }, mesh = {Adult ; Aged ; Antibodies/cerebrospinal fluid ; *Antibody Formation ; *Bacterial Proteins ; Blotting, Western ; Chaperonin 60 ; *Chaperonins ; Female ; Heat-Shock Proteins/*immunology ; Humans ; Male ; Middle Aged ; Multiple Sclerosis/cerebrospinal fluid/*immunology ; Mycobacterium bovis/immunology ; Nervous System Diseases/cerebrospinal fluid/immunology ; }, abstract = {The expression of heat shock proteins (hsp) within the target organ is implicated in the pathogenesis of a number of diseases of suspected autoimmune etiology, including MS. To pursue the potential role of a humoral response to the hsp 60/65 kd family in MS, we studied serum and CSF by Western blotting using recombinant Mycobacterium bovis hsp 65 and human hsp 60 as antigens and compared the findings with samples from patients with other neurologic diseases (OND). Analysis of the IgG response in CSF from 18 patients with MS indicated moderate reactivity in 10 cases and no reactivity in eight. In the OND group, reactivity was found in the CSF from one of two patients with Parkinson's disease, four of four Alzheimer's disease patients, and two of two patients with amyotrophic lateral sclerosis. CSF samples from seven of seven patients with subacute sclerosing panencephalitis were negative, as were samples from two normal subjects. There was no reactivity in CSF from two Huntington's disease patients. We conclude that antibodies reactive with hsp 60/65 are present in CSF of some MS patients but are also present in a number of chronic neurodegenerative conditions. The findings indicate that a humoral response to hsp 60/65 in the CSF is not specific for MS.}, }
@article {pmid8229070, year = {1993}, author = {Martin, JE and Swash, M and Mather, K and Leigh, PN}, title = {Expression of the human groEL stress-protein homologue in the brain and spinal cord.}, journal = {Journal of the neurological sciences}, volume = {118}, number = {2}, pages = {202-206}, doi = {10.1016/0022-510x(93)90111-b}, pmid = {8229070}, issn = {0022-510X}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Adolescent ; Adult ; Aged ; Antibodies, Monoclonal ; Astrocytes/metabolism ; Brain/pathology ; Brain Chemistry/*physiology ; Child ; Female ; Heat-Shock Proteins/*biosynthesis/immunology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mitochondria/metabolism ; Nervous System Diseases/pathology ; Neurons/metabolism ; Oligodendroglia/metabolism ; Spinal Cord/*metabolism/pathology ; }, abstract = {A monoclonal antibody (ML30), previously shown to identify a human mitochondrial protein epitope homologous with the groEL heat-shock protein of bacteria (hsp60), was used in an immunohistochemical survey of the central nervous system in patients dying with no evidence of neurological disease and in tissue from patients dying with various neurological disorders. Staining was performed on frozen tissue sections and on formalin fixed, paraffin embedded tissue. Astrocytes in all areas showed a strong pattern of punctate granular staining, which was increased in astrocytes showing reactive changes. Oligodendrocytes stained lightly in a diffuse granular pattern as did most neurons. Ependymal cells showed apical granular positivity. Expression of the hsp60 epitope recognised by ML30 was not seen in ubiquitinated inclusion bodies in motor neuron disease, neurofibrillary tangles in Alzheimer's disease or Lewy bodies in Parkinson's disease. The epitope recognised by ML30 was stable after formalin fixation and in post mortem tissue up to 96 h after death. Expression of the human groEL stress-protein homologue in brain and spinal cord is consistent with a mitochondrial location and may provide a morphological indicator of the functional or metabolic state of cells, especially glial cells.}, }
@article {pmid8714148, year = {1996}, author = {Khanna, N and Shankar, SK and Chandramuki, A and Jagannath, C}, title = {Immunohistochemical study of the expression of human groEL-stress protein in human nervous tissue.}, journal = {The Indian journal of medical research}, volume = {103}, number = {}, pages = {103-111}, pmid = {8714148}, issn = {0971-5916}, mesh = {Animals ; Brain Chemistry/*physiology ; Chaperonin 60/*analysis ; Gerbillinae ; Humans ; Immunohistochemistry ; Nerve Tissue Proteins/*analysis ; Rats ; Saimiri ; Spinal Cord/*chemistry ; }, abstract = {Monoclonal antibody (ML-30) directed against 65 kDa stress protein of mycobacteria, is shown to identify human cellular protein homologous with the groEL heat shock protein in many prokaryotes. Immunohistochemical survey of nervous tissue, both central and peripheral, from patients dying of various inflammatory, degenerative and neoplastic conditions and from experimental animals, using this antibody showed punctate granular staining of the cells to a variable degree. The astrocytes showed strong immunolabelling. The normal neurons and oligodendroglia stained variably, while abnormal neurons were darkly labelled. Ependymal cells showed apical granular positivity. The ubiquitinated inclusion bodies in amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease were not recognised by the ML-30 antibody. In diseased and stressed nervous tissue from experimental animals, the expression of the ML-30 recognisable stress protein was variable. The epitope recognised by ML-30 was found stable in postmortem tissues collected up to 36 h after death and processed for paraffin sectioning, after fixation in formalin for many years. Enhanced expression of the human groEL stress protein homologue in mammalian nervous tissue following various forms of stress may play a role in modulating the extent of tissue damage by autoimmune mechanism because of its high immunogenic nature and constitutive presence in the cells.}, }
@article {pmid8841663, year = {1996}, author = {Bergmann, M and Kuchelmeister, K and Schmid, KW and Kretzschmar, HA and Schröder, R}, title = {Different variants of frontotemporal dementia: a neuropathological and immunohistochemical study.}, journal = {Acta neuropathologica}, volume = {92}, number = {2}, pages = {170-179}, doi = {10.1007/s004010050505}, pmid = {8841663}, issn = {0001-6322}, mesh = {Adult ; Aged ; Atrophy/immunology/pathology ; Dementia/*immunology/*pathology ; Frontal Lobe/*immunology/*pathology ; Humans ; Immunohistochemistry ; Middle Aged ; Motor Neuron Disease/immunology/pathology ; Temporal Lobe/*immunology/*pathology ; }, abstract = {Histological and immunohistochemical findings in 20 cases of frontotemporal dementias-8 cases of dementia of frontal lobe type (DFT), 7 cases of Pick's disease (PD), and 5 cases of motor neuron disease with dementia (MND/D)-are presented. Common features of all three syndromes were: frontotemporal atrophy, involvement of subcortical nuclei, and swollen chromatolytic cells. Ubiquitin (Ub)-positive and tau-negative inclusions in cortical, hippocampal, and motor neurons were found in MND/D and DFT cases, suggesting a common pathogenesis of MND/D and DFT. MND/D showed the same cytoskeletal alterations in motor nuclei as MND without dementia: Bunina bodies and skein-like, Ub-positive inclusions. DFT differed from PD in the preponderance of histopathological changes in upper cortical layers, the sparseness of chromatolytic cells, and the absence of tau-positive Pick bodies (PBs). There were, however, two transitional cases showing Pick-type histology but no PBs, thus linking DFT and PD. PBs expressed chromogranin B and secretoneurin strongly, but chromogranin A only weakly. They were negative for the 70-kDa heat-shock protein, metallothionein, and glutathione-S-transferase.}, }
@article {pmid9273821, year = {1997}, author = {Kato, S and Hayashi, H and Nakashima, K and Nanba, E and Kato, M and Hirano, A and Nakano, I and Asayama, K and Ohama, E}, title = {Pathological characterization of astrocytic hyaline inclusions in familial amyotrophic lateral sclerosis.}, journal = {The American journal of pathology}, volume = {151}, number = {2}, pages = {611-620}, pmid = {9273821}, issn = {0002-9440}, mesh = {Adult ; Aged ; Amyotrophic Lateral Sclerosis/genetics/*pathology ; Astrocytes/*pathology/ultrastructure ; Brain/pathology/ultrastructure ; Female ; Humans ; Hyalin/*ultrastructure ; Immunohistochemistry ; Lewy Bodies/*ultrastructure ; Male ; Microscopy, Electron ; Middle Aged ; Spinal Cord/pathology/ultrastructure ; }, abstract = {To clarify the pathological characteristics of astrocytic hyaline inclusions (Ast-HIs) in patients with familial amyotrophic lateral sclerosis (FALS) with neuronal Lewy-body-like hyaline inclusions (LBHIs), eight autopsies on members of four different families, including two long-term surviving patients with clinical courses of over 10 years, were analyzed. Ast-HIs were found only in the two long-term surviving patients who belonged to different families and to different races. Ast-HIs were ultrastructurally composed of 15- to 25-nm granule-coated fibrils that had immunoreactivities to superoxide dismutase 1 (SOD1) and ubiquitin. Approximately 50% of the Ast-HIs expressed alpha B-crystallin, metallothionein, glutamine synthetase, and tubulin (alpha and beta) at various intensities. Some Ast-HIs reacted with antibodies to tau protein, S-100 protein, and heat shock protein 27. The Ast-HIs were not stained for glial fibrillary acidic protein. Our results suggest a cooperative role of superoxide dismutase 1, ubiquitin, and cytoskeletal proteins in the formation of granule-coated fibrils (namely, Ast-HIs) and provide evidence that Ast-HIs are formed in certain long-surviving familial amyotrophic lateral sclerosis patients with neuronal Lewy-body-like hyaline inclusions.}, }
@article {pmid9469581, year = {1998}, author = {Ariga, T and Jarvis, WD and Yu, RK}, title = {Role of sphingolipid-mediated cell death in neurodegenerative diseases.}, journal = {Journal of lipid research}, volume = {39}, number = {1}, pages = {1-16}, pmid = {9469581}, issn = {0022-2275}, support = {CA-09380/CA/NCI NIH HHS/United States ; NS-11853/NS/NINDS NIH HHS/United States ; NS-26994/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; *Apoptosis ; Gangliosides/physiology ; Glycolipids/physiology ; Humans ; Lipids/physiology ; Neurodegenerative Diseases/*pathology ; Second Messenger Systems ; Sphingolipids/*physiology ; }, abstract = {The metazoan nervous system gives rise intradevelopmentally to many more neurons than ultimately survive in the adult. Such excess cells are eliminated through programmed cell death or apoptosis. As is true for cells of other lineages, neuronal survival is sustained by an array of growth factors, such that withdrawal of neurotrophic support results in apoptotic cell death. Apoptosis is therefore believed to represent a beneficial process essential to normal development of central and peripheral nervous system (CNS and PNS) structures. Although the initiation of neuronal apoptosis in response to numerous extracellular agents has been widely reported, the regulatory mechanisms underlying this mode of cell death remain incompletely understood. In recent years, the contribution of lipid-dependent signaling systems, such as the sphingomyelin pathway, to regulation of cell survival has received considerable attention, leading to the identification of lethal functions for the lipid effectors ceramide and sphingosine in both normal and pathophysiological conditions. Moreover, the apoptotic capacities of several cytotoxic receptor systems (e.g., CD120a, CD95) and many environmental stresses (e.g., ionizing radiation, heat-shock, oxidative stress) are now known to derive from the activation of multiple signaling cascades by ceramide or, under some circumstances, by sphingosine. Inappropriate initiation of apoptosis has been proposed to underlie the progressive neuronal attrition associated with various neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and other neurological disorders that are characterized by the gradual loss of specific populations of neurons. In such pathophysiological states, neuronal cell death can result in specific disorders of movement and diverse impairments of CNS and PNS function. In some autoimmune neurological diseases such as Guillain-Barré syndrome, demyelinating polyneuropathy, and motoneuron disease, persistent immunological attack of microvascular endothelial cells by glycolipid-directed autoantibodies may lead to extensive cellular damages, resulting in increased permeability across brain-nerve barrier (BNB) and/or blood-brain barrier (BBB).}, }
@article {pmid9930742, year = {1999}, author = {Bruening, W and Roy, J and Giasson, B and Figlewicz, DA and Mushynski, WE and Durham, HD}, title = {Up-regulation of protein chaperones preserves viability of cells expressing toxic Cu/Zn-superoxide dismutase mutants associated with amyotrophic lateral sclerosis.}, journal = {Journal of neurochemistry}, volume = {72}, number = {2}, pages = {693-699}, doi = {10.1046/j.1471-4159.1999.0720693.x}, pmid = {9930742}, issn = {0022-3042}, support = {NIHRO1/HR/NHLBI NIH HHS/United States ; }, mesh = {3T3 Cells ; Amyotrophic Lateral Sclerosis/*metabolism ; Animals ; Cell Survival/physiology ; Chaperonins/*metabolism ; Gene Expression Regulation, Enzymologic ; Gene Transfer Techniques ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Heat-Shock Response/physiology ; Humans ; Mice ; Mice, Transgenic ; Motor Neurons/cytology/enzymology ; Mutation/physiology ; Neuroprotective Agents/metabolism ; Spinal Cord/cytology ; Superoxide Dismutase/*genetics/metabolism ; Transfection ; Up-Regulation/physiology ; }, abstract = {Mutations in the Cu/Zn-superoxide dismutase (SOD-1) gene underlie some familial cases of amyotrophic lateral sclerosis, a neurodegenerative disorder characterized by loss of cortical, brainstem, and spinal motor neurons. We present evidence that SOD-1 mutants alter the activity of molecular chaperones that aid in proper protein folding and targeting of abnormal proteins for degradation. In a cultured cell line (NIH 3T3), resistance to mutant SOD-1 toxicity correlated with increased overall chaperoning activity (measured by the ability of cytosolic extracts to prevent heat denaturation of catalase) as well as with up-regulation of individual chaperones/stress proteins. In transgenic mice expressing human SOD-1 with the G93A mutation, chaperoning activity was decreased in lumbar spinal cord but increased or unchanged in clinically unaffected tissues. Increasing the level of the stress-inducible chaperone 70-kDa heat shock protein by gene transfer reduced formation of mutant SOD-containing proteinaceous aggregates in cultured primary motor neurons expressing G93A SOD-1 and prolonged their survival. We propose that insufficiency of molecular chaperones may be directly involved in loss of motor neurons in this disease.}, }
@article {pmid11278741, year = {2001}, author = {Shinder, GA and Lacourse, MC and Minotti, S and Durham, HD}, title = {Mutant Cu/Zn-superoxide dismutase proteins have altered solubility and interact with heat shock/stress proteins in models of amyotrophic lateral sclerosis.}, journal = {The Journal of biological chemistry}, volume = {276}, number = {16}, pages = {12791-12796}, doi = {10.1074/jbc.M010759200}, pmid = {11278741}, issn = {0021-9258}, mesh = {3T3 Cells ; Amino Acid Substitution ; Animals ; Disease Models, Animal ; Heat-Shock Proteins/chemistry/*metabolism ; Humans ; Isoenzymes/chemistry/genetics/metabolism ; Mice ; Mice, Transgenic ; Motor Neuron Disease/*enzymology/*genetics ; Mutagenesis, Site-Directed ; Recombinant Proteins/chemistry/metabolism ; Solubility ; Superoxide Dismutase/*chemistry/genetics/*metabolism ; Transfection ; }, abstract = {Mutations in the Cu/Zn-superoxide dismutase (SOD-1) gene are responsible for a familial form of amyotrophic lateral sclerosis. In humans and experimental models, death of motor neurons is preceded by formation of cytoplasmic aggregates containing mutant SOD-1 protein. In our previous studies, heat shock protein 70 (HSP70) prolonged viability of cultured motor neurons expressing mutant human SOD-1 and reduced formation of aggregates. In this paper, we report that mutant SOD-1 proteins have altered solubility in cells relative to wild-type SOD-1 and can form a direct association with HSP70 and other stress proteins. Whereas wild-type human and endogenous mouse SOD-1 were detergent-soluble, a portion of mutant SOD-1 was detergent-insoluble in protein extracts of NIH3T3 transfected with SOD-1 gene constructs, spinal cord cultures established from G93A SOD-1 transgenic mouse embryos, and lumbar spinal cord from adult G93A transgenic mice. A direct association of HSP70, HSP40, and alphaB-crystallin with mutant SOD-1 (G93A or G41S), but not wild-type or endogenous mouse SOD-1, was demonstrated by coimmunoprecipitation. Mutant SOD-1.HSP70 complexes were predominantly in the detergent-insoluble fraction. However, only a small percentage of total cellular mutant SOD-1 was detergent-insoluble, suggesting that mutation-induced alteration of protein conformation may not in itself be sufficient for direct interaction with heat shock proteins.}, }
@article {pmid11458278, year = {2001}, author = {Gáti, I and Leel-Ossy, L}, title = {Heat shock protein 60 in corpora amylacea.}, journal = {Pathology oncology research : POR}, volume = {7}, number = {2}, pages = {140-144}, pmid = {11458278}, issn = {1219-4956}, mesh = {Astrocytes/chemistry ; Biomarkers ; Brain/*pathology ; *Brain Chemistry ; Brain Diseases/*metabolism/pathology ; Brain Neoplasms/chemistry/pathology ; Cerebral Infarction/metabolism/pathology ; Chaperonin 60/*analysis ; Encephalomyelitis, Acute Disseminated/metabolism/pathology ; Glioblastoma/chemistry/pathology ; Humans ; Inclusion Bodies/*chemistry ; Motor Neuron Disease/metabolism/pathology ; Multiple Sclerosis/metabolism/pathology ; Polysaccharides/analysis ; }, abstract = {Heat shock protein 60 representation in the corpora amylacea of the brain was investigated in five different neurological diseases. In the cases with cerebral infarct, amyotrophic lateral sclerosis, multiple sclerosis, acute disseminated encephalomyelitis and primary tumors of the nervous system the corpora amylacea showed similar appearance with strong HSP-60 positivity in all investigated disorders at the predilection sites. In the inflammatory diseases, besides corpora amylacea, several cellular elements exhibited HSP-60 immunostaining too. In these cases, the widespread HSP-60 immunoreactivity associated with relative moderate corpora amylacea production as compared to other diseases. From this contradiction we concluded the corpora amylacea participate in the cellular stress reaction but stress protein synthesis certainly is not the primary event in corpora amylacea formation. In the development of the corpora amylacea the incipient process is most probably degenerative in nature, which later on is accompanied by stress protein synthesis and slow growing of these round structures designated for a protective role in the brain. However, the role of the stress protein synthesis in the corpora amylacea formation and growth was not unequivocally answered in this study. It is necessary to perform further comparative investigations of the stress protein representation and corpora amylacea formation in different diseases which may help in discovering useful pathogenetic data and the biological role of this degenerative structure.}, }
@article {pmid11679167, year = {2001}, author = {Costa, V and Moradas-Ferreira, P}, title = {Oxidative stress and signal transduction in Saccharomyces cerevisiae: insights into ageing, apoptosis and diseases.}, journal = {Molecular aspects of medicine}, volume = {22}, number = {4-5}, pages = {217-246}, doi = {10.1016/s0098-2997(01)00012-7}, pmid = {11679167}, issn = {0098-2997}, mesh = {Aging/*metabolism ; Animals ; *Apoptosis ; Disease ; Humans ; *Oxidative Stress ; Saccharomyces cerevisiae/*metabolism/physiology ; *Signal Transduction ; }, abstract = {In yeast, as in higher eukaryotes, reactive oxygen species are produced as normal by-products of cellular metabolism. Under physiological conditions, the cell defence mechanisms are able to avoid molecular damages. This balance is disturbed when yeast cells are exposed to diverse environmental stress conditions, such as the presence of oxidants, heat shock, ethanol and metal ions. The increased production of reactive oxygen species is sensed by the cell, leading to the induction of defence mechanisms - the oxidative stress response. The present review discusses the mechanisms by which reactive oxygen species are sensed and the signalling pathways that are coupled with changes in genomic expression programs. Yeast has been used as an eukaryotic cell system to characterise the molecular mechanisms underlying the oxidative stress response. Furthermore, yeast has been utilised to elucidate the role of oxidative stress in ageing, apoptosis, and diseases, such as familial amyotrophic lateral sclerosis and Friedreich's ataxia.}, }
@article {pmid12032289, year = {2002}, author = {Kondo, M and Shibata, T and Kumagai, T and Osawa, T and Shibata, N and Kobayashi, M and Sasaki, S and Iwata, M and Noguchi, N and Uchida, K}, title = {15-Deoxy-Delta(12,14)-prostaglandin J(2): the endogenous electrophile that induces neuronal apoptosis.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {99}, number = {11}, pages = {7367-7372}, pmid = {12032289}, issn = {0027-8424}, mesh = {Adult ; Aged ; Apoptosis/*drug effects ; Female ; Flow Cytometry ; Gene Expression Regulation/drug effects ; Genes, p53 ; Humans ; Immunologic Factors/*pharmacology ; Male ; Middle Aged ; Motor Neuron Disease/pathology ; Motor Neurons/drug effects/pathology/physiology ; Neuroblastoma ; Neurons/cytology/*physiology ; Oligonucleotide Array Sequence Analysis ; Prostaglandin D2/*analogs & derivatives/analysis/*pharmacology ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53/genetics ; }, abstract = {Prostaglandin D(2) (PGD(2)), a major cyclooxygenase product in a variety of tissues and cells, readily undergoes dehydration to yield the bioactive cyclopentenone-type PGs of the J(2)-series, such as 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)). The observation that the level of 15d-PGJ(2) increased in the tissue cells from patients with sporadic amyotrophic lateral sclerosis suggested that the formation of 15d-PGJ(2) may be closely associated with neuronal cell death during chronic inflammatory processes. In vitro experiments using SH-SY5Y human neuroblastoma cells revealed that 15d-PGJ(2) induced apoptotic cell death. An oligonucleotide microarray analysis demonstrated that, in addition to the heat shock-responsive and redox-responsive genes, the p53-responsive genes, such as gadd45, cyclin G1, and cathepsin D, were significantly up-regulated in the cells treated with 15d-PGJ(2). Indeed, the 15d-PGJ(2) induced accumulation and phosphorylation of p53, which was accompanied by a preferential redistribution of the p53 protein in the nuclei of the cells and by a time-dependent increase in p53 DNA binding activity, suggesting that p53 accumulated in response to the treatment with 15d-PGJ(2) was functional. The 15d-PGJ(2)-induced accumulation of p53 resulted in the activation of a death-inducing caspase cascade mediated by Fas and the Fas ligand.}, }
@article {pmid12060716, year = {2002}, author = {Okado-Matsumoto, A and Fridovich, I}, title = {Amyotrophic lateral sclerosis: a proposed mechanism.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {99}, number = {13}, pages = {9010-9014}, pmid = {12060716}, issn = {0027-8424}, support = {R01 DK059868/DK/NIDDK NIH HHS/United States ; 1R01DK59868-01/DK/NIDDK NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/enzymology/*physiopathology ; Animals ; Humans ; Mice ; Mitochondria, Liver/enzymology ; Mitochondrial Swelling ; Mutation ; Superoxide Dismutase/genetics/metabolism ; }, abstract = {Missense mutations in Cu,Zn-superoxide dismutase (SOD1) account for approximately 20% of familial amyotrophic lateral sclerosis (FALS) through some, as yet undefined, toxic gain of function that leads to gradual death of motor neurons. Mitochondrial swelling and vacuolization are early signs of incipient motor neuron death in FALS. We previously reported that SOD1 exists in the intermembrane space of mitochondria. Herein, we demonstrate that the entry of SOD1 into mitochondria depends on demetallation and that heat shock proteins (Hsp70, Hsp27, or Hsp25) block the uptake of the FALS-associated mutant SOD1 (G37R, G41D, or G93A), while having no effect on wild-type SOD1. The binding of mutant SOD1 to Hsps in the extract of neuroblastoma cells leads to formation of sedimentable aggregates. Many antiapoptotic effects of Hsps have been reported. We now propose that this binding of Hsps to mutant forms of a protein abundant in motor neurons, such as SOD1, makes Hsps unavailable for their antiapoptotic functions and leads ultimately to motor neuron death. It also appears that the Hsp-SOD1 complex recruits other proteins present in the neuroblastoma cell and presumably in motor neurons to form sedimentable aggregates.}, }
@article {pmid12125078, year = {2002}, author = {Kikuchi, S and Shinpo, K and Takeuchi, M and Tsuji, S and Yabe, I and Niino, M and Tashiro, K}, title = {Effect of geranylgeranylaceton on cellular damage induced by proteasome inhibition in cultured spinal neurons.}, journal = {Journal of neuroscience research}, volume = {69}, number = {3}, pages = {373-381}, doi = {10.1002/jnr.10298}, pmid = {12125078}, issn = {0360-4012}, mesh = {Acetylcysteine/adverse effects/*analogs & derivatives ; Animals ; Cell Survival/drug effects ; Cells, Cultured ; Cysteine Endopeptidases/*metabolism ; Cysteine Proteinase Inhibitors/adverse effects ; Diterpenes/*pharmacology ; Dose-Response Relationship, Drug ; Glutathione/drug effects/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Immunoblotting ; Immunohistochemistry ; Mitochondria/drug effects/metabolism ; Multienzyme Complexes/*antagonists & inhibitors/*metabolism ; Neurons/drug effects/*metabolism ; Neuroprotective Agents/*pharmacology ; Oligopeptides/adverse effects ; Proteasome Endopeptidase Complex ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species/metabolism ; Spinal Cord/cytology/drug effects/*metabolism ; Thioredoxins/metabolism ; Time Factors ; }, abstract = {We investigated the effect of two proteasome inhibitors, lactacystin and epoxomicin, on cultured spinal cord neurons. The incubation of spinal neurons with proteasome inhibitors for 24 hr induced neurotoxicity in a dose-dependent manner. We found motor neurons to be more vulnerable to proteasome-induced neurotoxicity than nonmotor neurons. The staining of cell bodies in treated motor neurons was markedly disrupted and showed characteristic granular patterns. Proteasome-induced neurotoxicity is accompanied by apoptotic nuclear changes, posttranslational modification of the cellular proteins, generation of intracellular free radicals, reduction in the amount of reduced glutathione, and mitochondrial dysfunction. Neurotoxicity was reduced by the administration of low concentrations (1-100 nM) of geranylgeranylacetone (GGA), which is widely used as an antiulcer drug, although higher concentrations of this drug produced neurotoxicity in spinal cord neurons. GGA was found to induce the expression of heat shock protein 70 as well as thioredoxin, which may partly contribute to the protective effect of GGA. These data suggest that the inhibition of proteasome may play a role in the mechanism of neurodegenerative diseases of the spinal cord, such as amyotrophic lateral sclerosis, and that the use of GGA may be effective in the treatment of these conditions.}, }
@article {pmid12213295, year = {2002}, author = {Takeuchi, H and Kobayashi, Y and Yoshihara, T and Niwa, J and Doyu, M and Ohtsuka, K and Sobue, G}, title = {Hsp70 and Hsp40 improve neurite outgrowth and suppress intracytoplasmic aggregate formation in cultured neuronal cells expressing mutant SOD1.}, journal = {Brain research}, volume = {949}, number = {1-2}, pages = {11-22}, doi = {10.1016/s0006-8993(02)02568-4}, pmid = {12213295}, issn = {0006-8993}, mesh = {Amyotrophic Lateral Sclerosis/metabolism ; Baculoviridae ; Blotting, Western ; Cell Culture Techniques ; Cell Death/genetics ; Cytoplasm/*metabolism/ultrastructure ; Fluorescent Antibody Technique ; Gene Expression Regulation, Enzymologic ; HSP40 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins/biosynthesis/*metabolism ; Heat-Shock Proteins/biosynthesis/*metabolism ; Humans ; Microscopy, Confocal ; *Mutation ; *Neurites/metabolism ; Superoxide Dismutase/*genetics ; Superoxide Dismutase-1 ; Transfection ; Up-Regulation ; }, abstract = {Mutations of the superoxide dismutase 1 (SOD1) gene cause familial amyotrophic lateral sclerosis (FALS). Intracytoplasmic aggregate formation consisting of mutant SOD1 is the histological hallmark of FALS. Since a previous report revealed that Hsp70 reduced aggregate formation and cell death in a cell model of FALS, here we examined the combined effects of Hsp70 and its cofactor, Hsp40, on a cell model of FALS. The combination of Hsp70 and Hsp40 reduced intracytoplasmic aggregates and markedly improved neurite outgrowth. They also prevented cell death to a relatively lesser extent. Neurite outgrowth was recognized almost exclusively in the cells without intracytoplasmic aggregates. Hsp70 and Hsp40 were upregulated in cells expressing mutant SOD1, and were colocalized with intracytoplasmic aggregates of mutant SOD1. These findings suggest that heat shock proteins (HSPs) promote neurite outgrowth by suppressing intracytoplasmic aggregate formation and restoring cellular dysfunctions. This is the first demonstration that overexpression of HSPs improved neurite outgrowth as it suppressed intracytoplasmic aggregate formation and cell death in a cultured neuronal cell model of FALS. These findings may provide a basis for the utilization of HSPs in developing a treatment for FALS.}, }
@article {pmid12430713, year = {2002}, author = {Vleminckx, V and Van Damme, P and Goffin, K and Delye, H and Van Den Bosch, L and Robberecht, W}, title = {Upregulation of HSP27 in a transgenic model of ALS.}, journal = {Journal of neuropathology and experimental neurology}, volume = {61}, number = {11}, pages = {968-974}, doi = {10.1093/jnen/61.11.968}, pmid = {12430713}, issn = {0022-3069}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*pathology/*physiopathology ; Animals ; Blotting, Western ; Disease Models, Animal ; HSP70 Heat-Shock Proteins/metabolism ; *Heat-Shock Proteins ; Immunohistochemistry ; Mice ; Mice, Transgenic ; Molecular Chaperones ; Mutation/genetics ; Neoplasm Proteins/*analysis ; Neuroglia/pathology ; Neurons/pathology ; Oxidative Stress/physiology ; Spinal Cord/*pathology/physiopathology ; Superoxide Dismutase/genetics ; Superoxide Dismutase-1 ; Up-Regulation/*physiology ; alpha-Crystallin B Chain/metabolism ; }, abstract = {Mutations of the SOD1 gene underlie 1 form of familial amyotrophic lateral sclerosis (ALS). Their pathogenic mechanism remains uncertain, but is thought to involve oxidative stress and abnormal protein aggregation, 2 processes known to induce heat shock proteins (HSPs). We studied the expression of 3 HSPs (alphaB-crystallin, HSP27, and HSP70) in transgenic mice overexpressing human mutant (G93A and G37R) SOD1, using a combination of immunohistochemistry and immunoblotting. Quantitative Western blot analysis demonstrated alphaB-crystallin and HSP27 to be upregulated in the spinal cord of mutant SOD1 mice compared to mice overexpressing wild-type SOD1. HSP70 levels were normal. Immunocytochemical studies of the ventral horn of the spinal cord demonstrated HSP27 to be localized in the nucleus of neurons and glial cells in presymptomatic and early symptomatic animals, where it often was punctate in pattern. In the later stages of the disease, HSP27 was predominantly present in the cytoplasm of reactive glial cells. The early nuclear localization was confirmed by Western blot analysis of spinal cord nuclear and cytoplasmic fractions. In contrast to HSP27, alphaB-crystallin was localized exclusively in the cytoplasm of reactive glial cells.}, }
@article {pmid12537588, year = {2002}, author = {Fritzsche, M}, title = {Geographical and seasonal correlation of multiple sclerosis to sporadic schizophrenia.}, journal = {International journal of health geographics}, volume = {1}, number = {1}, pages = {5}, pmid = {12537588}, issn = {1476-072X}, abstract = {BACKGROUND: Clusters by season and locality reveal a striking epidemiological overlap between sporadic schizophrenia and multiple sclerosis (MS). As the birth excesses of those individuals who later in life develop schizophrenia mirror the seasonal distribution of Ixodid ticks, a meta analysis has been performed between all neuropsychiatric birth excesses including MS and the epidemiology of spirochaetal infectious diseases. RESULTS: The prevalence of MS and schizophrenic birth excesses entirely spares the tropical belt where human treponematoses are endemic, whereas in more temperate climates infection rates of Borrelia garinii in ticks collected from seabirds match the global geographic distribution of MS. If the seasonal fluctuations of Lyme borreliosis in Europe are taken into account, the birth excesses of MS and those of schizophrenia are nine months apart, reflecting the activity of Ixodes ricinus at the time of embryonic implantation and birth. In America, this nine months' shift between MS and schizophrenic births is also reflected by the periodicity of Borrelia burgdorferi transmitting Ixodes pacificus ticks along the West Coast and the periodicity of Ixodes scapularis along the East Coast. With respect to Ixodid tick activity, amongst the neuropsychiatric birth excesses only amyotrophic lateral sclerosis (ALS) shows a similar seasonal trend. CONCLUSION: It cannot be excluded at present that maternal infection by Borrelia burgdorferi poses a risk to the unborn. The seasonal and geographical overlap between schizophrenia, MS and neuroborreliosis rather emphasises a causal relation that derives from exposure to a flagellar virulence factor at conception and delivery. It is hoped that the pathogenic correlation of spirochaetal virulence to temperature and heat shock proteins (HSP) might encourage a new direction of research in molecular epidemiology.}, }
@article {pmid12657679, year = {2003}, author = {Adachi, H and Katsuno, M and Minamiyama, M and Sang, C and Pagoulatos, G and Angelidis, C and Kusakabe, M and Yoshiki, A and Kobayashi, Y and Doyu, M and Sobue, G}, title = {Heat shock protein 70 chaperone overexpression ameliorates phenotypes of the spinal and bulbar muscular atrophy transgenic mouse model by reducing nuclear-localized mutant androgen receptor protein.}, journal = {The Journal of neuroscience : the official journal of the Society for Neuroscience}, volume = {23}, number = {6}, pages = {2203-2211}, pmid = {12657679}, issn = {1529-2401}, mesh = {Animals ; Blotting, Western ; Cell Nucleus/*metabolism/pathology ; Crosses, Genetic ; Disease Models, Animal ; Disease Progression ; Gene Expression ; HSP70 Heat-Shock Proteins/*biosynthesis/genetics ; Humans ; Immunohistochemistry ; Macromolecular Substances ; Male ; Mice ; Mice, Transgenic ; Molecular Chaperones/*biosynthesis/genetics ; Motor Activity/genetics ; Muscular Atrophy, Spinal/genetics/pathology/*physiopathology ; Mutation ; Phenotype ; Receptors, Androgen/genetics/*metabolism ; Trinucleotide Repeat Expansion/genetics ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is an inherited motor neuron disease caused by the expansion of the polyglutamine (polyQ) tract within the androgen receptor (AR). The nuclear inclusions consisting of the mutant AR protein are characteristic and combine with many components of ubiquitin-proteasome and molecular chaperone pathways, raising the possibility that misfolding and altered degradation of mutant AR may be involved in the pathogenesis. We have reported that the overexpression of heat shock protein (HSP) chaperones reduces mutant AR aggregation and cell death in a neuronal cell model (Kobayashi et al., 2000). To determine whether increasing the expression level of chaperone improves the phenotype in a mouse model, we cross-bred SBMA transgenic mice with mice overexpressing the inducible form of human HSP70. We demonstrated that high expression of HSP70 markedly ameliorated the motor function of the SBMA model mice. In double-transgenic mice, the nuclear-localized mutant AR protein, particularly that of the large complex form, was significantly reduced. Monomeric mutant AR was also reduced in amount by HSP70 overexpression, suggesting the enhanced degradation of mutant AR. These findings suggest that HSP70 overexpression ameliorates SBMA phenotypes in mice by reducing nuclear-localized mutant AR, probably caused by enhanced mutant AR degradation. Our study may provide the basis for the development of an HSP70-related therapy for SBMA and other polyQ diseases.}, }
@article {pmid12843283, year = {2003}, author = {Batulan, Z and Shinder, GA and Minotti, S and He, BP and Doroudchi, MM and Nalbantoglu, J and Strong, MJ and Durham, HD}, title = {High threshold for induction of the stress response in motor neurons is associated with failure to activate HSF1.}, journal = {The Journal of neuroscience : the official journal of the Society for Neuroscience}, volume = {23}, number = {13}, pages = {5789-5798}, pmid = {12843283}, issn = {1529-2401}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism/pathology ; Animals ; Cells, Cultured ; Cysteine Endopeptidases/metabolism ; DNA-Binding Proteins/*metabolism ; Disease Models, Animal ; Enzyme Inhibitors/pharmacology ; *Gene Expression Regulation ; Genes, Reporter ; Glutamic Acid/toxicity ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Heat Shock Transcription Factors ; Heat-Shock Proteins/metabolism ; *Heat-Shock Response/physiology ; Hot Temperature ; Humans ; In Vitro Techniques ; Mice ; Mice, Transgenic ; Motor Neurons/drug effects/*metabolism/pathology ; Multienzyme Complexes/antagonists & inhibitors/metabolism ; Promoter Regions, Genetic ; Proteasome Endopeptidase Complex ; Spinal Cord/cytology/metabolism/pathology ; Superoxide Dismutase/biosynthesis/genetics ; Transcription Factors/metabolism ; }, abstract = {Heat shock protein 70 (Hsp70) protects cultured motor neurons from the toxic effects of mutations in Cu/Zn-superoxide dismutase (SOD-1), which is responsible for a familial form of the disease, amyotrophic lateral sclerosis (ALS). Here, the endogenous heat shock response of motor neurons was investigated to determine whether a high threshold for activating this protective mechanism contributes to their vulnerability to stresses associated with ALS. When heat shocked, cultured motor neurons failed to express Hsp70 or transactivate a green fluorescent protein reporter gene driven by the Hsp70 promoter, although Hsp70 was induced in glial cells. No increase in Hsp70 occurred in motor neurons after exposure to excitotoxic glutamate or expression of mutant SOD-1 with a glycine--> alanine substitution at residue 93 (G93A), nor was Hsp70 increased in spinal cords of G93A SOD-1 transgenic mice or sporadic or familial ALS patients. In contrast, strong Hsp70 induction occurred in motor neurons with expression of a constitutively active form of heat shock transcription factor (HSF)-1 or when proteasome activity was sufficiently inhibited to induce accumulation of an alternative transcription factor HSF2. These results indicate that the high threshold for induction of the stress response in motor neurons stems from an impaired ability to activate the main heat shock-stress sensor, HSF1.}, }
@article {pmid14636160, year = {2003}, author = {Wood, JD and Beaujeux, TP and Shaw, PJ}, title = {Protein aggregation in motor neurone disorders.}, journal = {Neuropathology and applied neurobiology}, volume = {29}, number = {6}, pages = {529-545}, doi = {10.1046/j.0305-1846.2003.00518.x}, pmid = {14636160}, issn = {0305-1846}, mesh = {Animals ; Disease Models, Animal ; Humans ; Inclusion Bodies/*pathology ; Motor Neuron Disease/genetics/*pathology ; Mutation ; Protein Conformation ; Proteins/chemistry/*metabolism ; Superoxide Dismutase/genetics ; }, abstract = {Toxicity associated with abnormal protein folding and protein aggregation are major hypotheses for neurodegeneration. This article comparatively reviews the experimental and human tissue-based evidence for the involvement of such mechanisms in neuronal death associated with the motor system disorders of X-linked spinobulbar muscular atrophy (SBMA; Kennedy's disease) and amyotrophic lateral sclerosis (ALS), especially disease related to mutations in the superoxide dismutase (SOD1) gene. Evidence from transgenic mouse, Drosophila and cell culture models of SBMA, in common with other trinucleotide repeat expansion disorders, show protein aggregation of the mutated androgen receptor, and intraneuronal accumulation of aggregated protein, to be obligate mechanisms. Strong experimental data link these phenomena with downstream biochemical events involving gene transcription pathways (CREB-binding protein) and interactions with protein chaperone systems. Manipulations of these pathways are already established in experimental systems of trinucleotide repeat disorders as potential beneficial targets for therapeutic activity. In contrast, the evidence for the role of protein aggregation in models of SOD1-linked familial ALS is less clear-cut. Several classes of intraneuronal inclusion body have been described, some of which are invariably present. However, the lack of understanding of the biochemical basis of the most frequent inclusion in sporadic ALS, the ubiquitinated inclusion, has hampered research. The toxicity associated with expression of mutant SOD1 has been intensively studied however. Abnormal protein aggregation and folding is the only one of the four major hypotheses for the mechanism of neuronal degeneration in this disorder currently under investigation (the others comprise oxidative stress, axonal transport and cytoskeletal dysfunctions, and glutamatergic excitotoxicity). Whilst hyaline inclusions, which are strongly immunoreactive to SOD1, are linked to degeneration in SOD1 mutant mouse models, the evidence from human tissue is less consistent and convincing. A role for mutant SOD1 aggregation in the mitochondrial dysfunction associated with ALS, and in potentially toxic interactions with heat shock proteins, both leading to apoptosis, are supported by some experimental data. Direct in vitro data on mutant SOD1 show evidence for spontaneous oligomerization, but the role of such oligomers remains to be elucidated, and therapeutic strategies are less well developed for this familial variant of ALS.}, }
@article {pmid15034571, year = {2004}, author = {Kieran, D and Kalmar, B and Dick, JR and Riddoch-Contreras, J and Burnstock, G and Greensmith, L}, title = {Treatment with arimoclomol, a coinducer of heat shock proteins, delays disease progression in ALS mice.}, journal = {Nature medicine}, volume = {10}, number = {4}, pages = {402-405}, doi = {10.1038/nm1021}, pmid = {15034571}, issn = {1078-8956}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/enzymology/pathology/physiopathology ; Animals ; Disease Progression ; Heat-Shock Proteins/*biosynthesis ; Humans ; Hydroxylamines/pharmacology/*therapeutic use ; Mice ; Mice, Transgenic ; Motor Neurons/pathology ; Mutation ; Superoxide Dismutase/genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative condition in which motoneurons of the spinal cord and motor cortex die, resulting in progressive paralysis. This condition has no cure and results in eventual death, usually within 1-5 years of diagnosis. Although the specific etiology of ALS is unknown, 20% of familial cases of the disease carry mutations in the gene encoding Cu/Zn superoxide dismutase-1 (SOD1). Transgenic mice overexpressing human mutant SOD1 have a phenotype and pathology that are very similar to that seen in human ALS patients. Here we show that treatment with arimoclomol, a coinducer of heat shock proteins (HSPs), significantly delays disease progression in mice expressing a SOD1 mutant in which glycine is substituted with alanine at position 93 (SOD1(G93A)). Arimoclomol-treated SOD1(G93A) mice show marked improvement in hind limb muscle function and motoneuron survival in the later stages of the disease, resulting in a 22% increase in lifespan. Pharmacological activation of the heat shock response may therefore be a successful therapeutic approach to treating ALS, and possibly other neurodegenerative diseases.}, }
@article {pmid15133611, year = {2004}, author = {Katsuno, M and Adachi, H and Tanaka, F and Sobue, G}, title = {Spinal and bulbar muscular atrophy: ligand-dependent pathogenesis and therapeutic perspectives.}, journal = {Journal of molecular medicine (Berlin, Germany)}, volume = {82}, number = {5}, pages = {298-307}, pmid = {15133611}, issn = {0946-2716}, mesh = {Animals ; Disease Models, Animal ; Humans ; Leuprolide/therapeutic use ; Ligands ; Mice ; Muscle, Skeletal/pathology ; Muscular Atrophy, Spinal/*drug therapy/etiology/*genetics ; Mutation ; Peptides/genetics ; Receptors, Androgen/*genetics/*metabolism ; Spinal Cord/pathology ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is a late-onset motor neuron disease characterized by proximal muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. SBMA exclusively affects males, while females are usually asymptomatic. The molecular basis of SBMA is the expansion of a trinucleotide CAG repeat, which encodes the polyglutamine (polyQ) tract in the first exon of the androgen receptor (AR) gene. The histopathological hallmark is the presence of nuclear inclusions containing mutant truncated ARs with expanded polyQ tracts in the residual motor neurons in the brainstem and spinal cord, as well as in some other visceral organs. The AR ligand, testosterone, accelerates AR dissociation from heat shock proteins and thus its nuclear translocation. Ligand-dependent nuclear accumulation of mutant ARs has been implicated in the pathogenesis of SBMA. Transgenic mice carrying the full-length human AR gene with an expanded polyQ tract demonstrate neuromuscular phenotypes, which are profound in males. Their SBMA-like phenotypes are rescued by castration, and aggravated by testosterone administration. Leuprorelin, an LHRH agonist that reduces testosterone release from the testis, inhibits nuclear accumulation of mutant ARs, resulting in the rescue of motor dysfunction in the male transgenic mice. However, flutamide, an androgen antagonist promoting nuclear translocation of the AR, yielded no therapeutic effect. The degradation and cleavage of the AR protein are also influenced by the ligand, contributing to the pathogenesis. Testosterone thus appears to be the key molecule in the pathogenesis of SBMA, as well as main therapeutic target of this disease.}, }
@article {pmid15198682, year = {2004}, author = {Urushitani, M and Kurisu, J and Tateno, M and Hatakeyama, S and Nakayama, K and Kato, S and Takahashi, R}, title = {CHIP promotes proteasomal degradation of familial ALS-linked mutant SOD1 by ubiquitinating Hsp/Hsc70.}, journal = {Journal of neurochemistry}, volume = {90}, number = {1}, pages = {231-244}, doi = {10.1111/j.1471-4159.2004.02486.x}, pmid = {15198682}, issn = {0022-3042}, mesh = {Amyotrophic Lateral Sclerosis/enzymology/genetics/*metabolism ; Animals ; Cells, Cultured ; Disease Models, Animal ; HSC70 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins/genetics/*metabolism ; Humans ; Inclusion Bodies/metabolism ; Macromolecular Substances ; Mice ; Mice, Transgenic ; Motor Neurons/metabolism ; Mutation ; Peptide Hydrolases/*metabolism ; *Proteasome Endopeptidase Complex ; Protein Subunits/metabolism ; Superoxide Dismutase/genetics/*metabolism ; Transfection ; Ubiquitin/*metabolism ; Ubiquitin-Protein Ligases/genetics/metabolism/*physiology ; }, abstract = {Over 100 mutants in superoxide dismutase 1 (SOD1) are reported in familial amyotrophic lateral sclerosis (ALS). However, the precise mechanism by which they are degraded through a ubiquitin-proteasomal pathway (UPP) remains unclear. Here, we report that heat-shock protein (Hsp) or heat-shock cognate (Hsc)70, and the carboxyl terminus of the Hsc70-interacting protein (CHIP), are involved in proteasomal degradation of mutant SOD1. Only mutant SOD1 interacted with Hsp/Hsc70 in vivo, and in vitro experiments revealed that Hsp/Hsc70 preferentially interacted with apo-SOD1 or dithiothreitol (DTT)-treated holo-SOD1, compared with metallated or oxidized forms. CHIP, a binding partner of Hsp/Hsc70, interacted only with mutant SOD1 and promoted its degradation. Both Hsp70 and CHIP promoted polyubiquitination of mutant SOD1-associated molecules, but not of mutant SOD1, indicating that mutant SOD1 is not a substrate of CHIP. Moreover, mutant SOD1-associated Hsp/Hsc70, a known substrate of CHIP, was polyubiquitinated in vivo, and polyubiquitinated Hsc70 by CHIP interacted with the S5a subunit of the 26S proteasome in vitro. Furthermore, CHIP was predominantly expressed in spinal neurons, and ubiquitinated inclusions in the spinal motor neurons of hSOD1(G93A) transgenic mice were CHIP-immunoreactive. Taken together, we propose a novel pathway in which ubiquitinated Hsp/Hsc70 might deliver mutant SOD1 to, and facilitate its degradation, at the proteasome.}, }
@article {pmid15245475, year = {2004}, author = {Maatkamp, A and Vlug, A and Haasdijk, E and Troost, D and French, PJ and Jaarsma, D}, title = {Decrease of Hsp25 protein expression precedes degeneration of motoneurons in ALS-SOD1 mice.}, journal = {The European journal of neuroscience}, volume = {20}, number = {1}, pages = {14-28}, doi = {10.1111/j.1460-9568.2004.03430.x}, pmid = {15245475}, issn = {0953-816X}, mesh = {Activating Transcription Factor 3 ; Adult ; Age Factors ; Aged ; Amyotrophic Lateral Sclerosis/genetics/*metabolism ; Animals ; Animals, Newborn ; Blotting, Western/methods ; Case-Control Studies ; Choline O-Acetyltransferase/metabolism ; Disease Models, Animal ; Female ; Gene Expression ; Gene Expression Regulation, Developmental ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/genetics/*metabolism ; Humans ; Immunohistochemistry/methods/statistics & numerical data ; In Situ Hybridization/methods ; Male ; Mice ; Mice, Transgenic ; Middle Aged ; Molecular Chaperones ; Motor Neurons/*metabolism ; Neoplasm Proteins/genetics/*metabolism ; Nerve Degeneration/genetics/*metabolism ; Postmortem Changes ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Spinal Cord/cytology/metabolism ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1 ; Transcription Factors/metabolism ; Ubiquitin/metabolism ; }, abstract = {We have investigated the expression of Hsp25, a heat shock protein constitutively expressed in motoneurons, in amyotrophic lateral sclerosis (ALS) mice that express G93A mutant SOD1 (G93A mice). Immunocytochemistry and Western blotting showed that a decrease of Hsp25 protein expression occurred in motoneurons of G93A mice prior to the onset of motoneuron death and muscle weakness. This decrease in Hsp25 expression also preceded the appearance of SOD1 aggregates as identified by cellulose acetate filtration and Western blot analysis. In contrast to Hsp25 protein levels, Hsp25 mRNA as determined by in situ hybridization and RT-PCR, remained unchanged. This suggests that the decrease in Hsp25 protein levels occurs post-transcriptionally. In view of the cytoprotective properties of Hsp25 and the temporal relationship between decreased Hsp25 expression and the onset of motoneuron death, it is feasible that reduced Hsp25 concentration contributes to the degeneration of motoneurons in G93A mice. These data are consistent with the idea that mutant SOD1 may reduce the availability of the protein quality control machinery in motoneurons.}, }
@article {pmid15341181, year = {2004}, author = {Calabrese, V and Boyd-Kimball, D and Scapagnini, G and Butterfield, DA}, title = {Nitric oxide and cellular stress response in brain aging and neurodegenerative disorders: the role of vitagenes.}, journal = {In vivo (Athens, Greece)}, volume = {18}, number = {3}, pages = {245-267}, pmid = {15341181}, issn = {0258-851X}, mesh = {Aging/*physiology ; Animals ; Brain/*growth & development ; Cell Death ; Cell Survival ; Humans ; Neurodegenerative Diseases/genetics/*physiopathology ; Nitric Oxide/*physiology ; Oxidative Stress/*physiology ; }, abstract = {Nitric oxide and other reactive nitrogen species appear to play crucial roles in the brain such as neuromodulation, neurotransmission and synaptic plasticity, but are also involved in pathological processes such as neurodegeneration and neuroinflammation. Acute and chronic inflammation result in increased nitrogen monoxide formation and nitrosative stress. It is now well documented that NO and its toxic metabolite, peroxynitrite, can inhibit components of the mitochondrial respiratory chain leading to cellular energy deficiency and, eventually, to cell death. Within the brain, the susceptibility of different brain cell types to NO and peroxynitrite exposure may be dependent on factors such as the intracellular reduced glutathione and cellular stress resistance signal pathways. Thus neurons, in contrast to astrocytes, appear particularly vulnerable to the effect of nitrosative stress. Evidence is now available to support this scenario for neurological disorders such as Alzheimer's disease, amyotrophic lateral sclerosis, Parkinson's disease, multiple sclerosis and Huntington's disease, but also in the brain damage following ischemia and reperfusion, Down's syndrome and mitochondrial encephalopathies. To survive different types of injuries, brain cells have evolved integrated responses, the so-called longevity assurance processes, composed of several genes termed vitagenes and including, among others, members of the HSP system, such as HSP70 and HSP32, to detect and control diverse forms of stress. In particular, HSP32, also known as heme oxygenase-1 (HO-1), has received considerable attention, as it has been recently demonstrated that HO-1 induction, by generating the vasoactive molecule carbon monoxide and the potent antioxidant bilirubin, could represent a protective system potentially active against brain oxidative injury. Increasing evidence suggests that the HO-1 gene is redox-regulated and its expression appears closely related to conditions of oxidative and nitrosative stress. An amount of experimental evidence indicates that increased rate of free radical generation and decreased efficiency of the reparative/degradative mechanisms, such as proteolysis, are factors that primarily contribute to age-related elevation in the level of oxidative stress and brain damage. Given the broad cytoprotective properties of the heat shock response there is now strong interest in discovering and developing pharmacological agents capable of inducing such a response. These findings have led to new perspectives in medicine and pharmacology, as molecules inducing this defense mechanism appear to be possible candidates for novel, cytoprotective strategies. Particularly, manipulation of endogenous cellular defense mechanisms such as the heat shock response, through nutritional antioxidants or pharmacological compounds, represents an innovative approach to therapeutic intervention in diseases causing tissue damage, such as neurodegeneration. Consistent with this notion, maintenance or recovery of the activity of vitagenes may possibly delay the aging process and decrease the occurrence of age-related diseases with resulting prolongation of a healthy life span.}, }
@article {pmid15465612, year = {2004}, author = {Yonekura, K and Yokota, S and Tanaka, S and Kubota, H and Fujii, N and Matsumoto, H and Chiba, S}, title = {Prevalence of anti-heat shock protein antibodies in cerebrospinal fluids of patients with Guillain-Barré syndrome.}, journal = {Journal of neuroimmunology}, volume = {156}, number = {1-2}, pages = {204-209}, doi = {10.1016/j.jneuroim.2004.07.017}, pmid = {15465612}, issn = {0165-5728}, mesh = {Adult ; Autoantibodies/*cerebrospinal fluid ; Female ; Guillain-Barre Syndrome/*cerebrospinal fluid/epidemiology/*immunology ; Heat-Shock Proteins/*cerebrospinal fluid/*immunology ; Humans ; Immunoglobulin G/cerebrospinal fluid ; Immunoglobulin M/cerebrospinal fluid ; Male ; Middle Aged ; Seroepidemiologic Studies ; }, abstract = {We examined antibodies against 10 heat shock proteins (HSPs) in cerebrospinal fluids (CSF) and sera from patients with Guillain-Barré syndrome (GBS). Significantly higher IgG antibody titers against HSP27, HSP60, HSP70 and HSP90 family, including mycobacterial HSP65 and Escherichia coli GroEL, were found in CSF from GBS patients as compared with motor neuron disease. Serum IgG antibodies against each HSP showed no difference between GBS patients and normal controls. GBS seems to be induced by reactive autoimmune responses frequently triggered by infections. The CSF antibodies against HSPs may modify the immune responses and/or cell-protective functions of HSPs in the pathophysiology of GBS.}, }
@article {pmid15509539, year = {2004}, author = {Strey, CW and Spellman, D and Stieber, A and Gonatas, JO and Wang, X and Lambris, JD and Gonatas, NK}, title = {Dysregulation of stathmin, a microtubule-destabilizing protein, and up-regulation of Hsp25, Hsp27, and the antioxidant peroxiredoxin 6 in a mouse model of familial amyotrophic lateral sclerosis.}, journal = {The American journal of pathology}, volume = {165}, number = {5}, pages = {1701-1718}, pmid = {15509539}, issn = {0002-9440}, support = {NS 36732/NS/NINDS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/*genetics ; Animals ; Antioxidants/*pharmacology ; Astrocytes/metabolism ; Blotting, Western ; Electrophoresis, Gel, Two-Dimensional ; *Gene Expression Regulation ; Golgi Apparatus/metabolism ; HSP27 Heat-Shock Proteins ; HeLa Cells ; Heat-Shock Proteins/*biosynthesis ; Humans ; Immunohistochemistry ; Immunoprecipitation ; Mass Spectrometry ; Mice ; Mice, Transgenic ; Microscopy, Fluorescence ; Microtubule Proteins/*biosynthesis/genetics/metabolism ; Microtubules/metabolism ; Models, Biological ; Molecular Chaperones ; Motor Neurons/metabolism ; Mutation ; Neoplasm Proteins/*biosynthesis ; Neurodegenerative Diseases/metabolism ; Neurons/metabolism ; Parkinson Disease/metabolism ; Peroxidases/*biosynthesis ; Peroxiredoxin VI ; Peroxiredoxins ; Phosphoproteins/*biosynthesis/genetics/metabolism ; Phosphorylation ; Plasmids/metabolism ; Protein Isoforms ; Protein Structure, Tertiary ; Spinal Cord/metabolism ; Stathmin ; Transfection ; *Up-Regulation ; }, abstract = {Gain-of-function mutations of the Cu/Zn superoxide dismutase (SOD1) gene cause dominantly inherited familial amyotrophic lateral sclerosis. The identification of differentially regulated proteins in spinal cords of paralyzed mice expressing SOD1(G93A) may contribute to understanding mechanisms of toxicity by mutant SOD1. Protein profiling showed dysregulation of Stathmin with a marked decrease of its most acidic and phosphorylated isoform, and up-regulation of heat shock proteins 25 and 27, peroxiredoxin 6, phosphatidylinositol transfer protein-alpha, apolipoprotein E, and ferritin heavy chain. Stathmin accumulated in the cytoplasm of 30% of spinal cord motor neurons with fragmented Golgi apparatus. Overexpression of Stathmin in HeLa cells was associated with collapse of microtubule networks and Golgi fragmentation. These results, together with the decrease of one Stathmin isoform, suggest a role of the protein in Golgi fragmentation. Mutant SOD1 co-precipitated and co-localized with Hsp25 in neurons and astrocytes. Mutant SOD1 may thus deprive cells of the anti-apoptotic and other protective activities of Hsp25. Astrocytes contained peroxiredoxin 6, a unique nonredundant antioxidant. The up-regulation of peroxiredoxin 6 probably constitutes a defense to oxidative stress induced by SOD1(G93A). Direct effects of SOD1(G93A) or sequential reactions triggered by the mutant may cause the protein changes.}, }
@article {pmid15699043, year = {2005}, author = {Casoni, F and Basso, M and Massignan, T and Gianazza, E and Cheroni, C and Salmona, M and Bendotti, C and Bonetto, V}, title = {Protein nitration in a mouse model of familial amyotrophic lateral sclerosis: possible multifunctional role in the pathogenesis.}, journal = {The Journal of biological chemistry}, volume = {280}, number = {16}, pages = {16295-16304}, doi = {10.1074/jbc.M413111200}, pmid = {15699043}, issn = {0021-9258}, support = {TCP01010/TI_/Telethon/Italy ; }, mesh = {Amino Acid Substitution ; Amyotrophic Lateral Sclerosis/etiology/*metabolism ; Animals ; Disease Models, Animal ; Mice ; Mice, Transgenic ; Mutation ; Nitrates/*metabolism ; Proteins/*metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spinal Cord/metabolism ; Superoxide Dismutase/*genetics/metabolism ; Superoxide Dismutase-1 ; }, abstract = {Multiple mechanisms have been proposed to contribute to amyotrophic lateral sclerosis (ALS) pathogenesis, including oxidative stress. Early evidence of a role for oxidative damage was based on the finding, in patients and murine models, of high levels of markers, such as free nitrotyrosine (NT). However, no comprehensive study on the protein targets of nitration in ALS has been reported. We found an increased level of NT immunoreactivity in spinal cord protein extracts of a transgenic mouse model of familial ALS (FALS) at a presymptomatic stage of the disease compared with age-matched controls. NT immunoreactivity is increased in the soluble fraction of spinal cord homogenates and is found as a punctate staining in motor neuron perikarya of presymptomatic FALS mice. Using a proteome-based strategy, we identified proteins nitrated in vivo, under physiological or pathological conditions, and compared their level of specific nitration. alpha- and gamma-enolase, ATP synthase beta chain, and heat shock cognate 71-kDa protein and actin were overnitrated in presymptomatic FALS mice. We identified by matrix-assisted laser desorption/ionization mass spectrometry 16 sites of nitration in proteins oxidized in vivo. In particular, alpha-enolase nitration at Tyr(43), target also of phosphorylation, brings additional evidence on the possible interference of nitration with phosphorylation. In conclusion, we propose that protein nitration may have a role in ALS pathogenesis, acting directly by inhibiting the function of specific proteins and indirectly interfering with protein degradation pathways and phosphorylation cascades.}, }
@article {pmid15749392, year = {2005}, author = {Perluigi, M and Fai Poon, H and Hensley, K and Pierce, WM and Klein, JB and Calabrese, V and De Marco, C and Butterfield, DA}, title = {Proteomic analysis of 4-hydroxy-2-nonenal-modified proteins in G93A-SOD1 transgenic mice--a model of familial amyotrophic lateral sclerosis.}, journal = {Free radical biology & medicine}, volume = {38}, number = {7}, pages = {960-968}, doi = {10.1016/j.freeradbiomed.2004.12.021}, pmid = {15749392}, issn = {0891-5849}, support = {AG-05119/AG/NIA NIH HHS/United States ; AG-10836/AG/NIA NIH HHS/United States ; NS044154/NS/NINDS NIH HHS/United States ; }, mesh = {Aldehydes/*analysis ; Amyotrophic Lateral Sclerosis/genetics/*metabolism ; Animals ; Disease Models, Animal ; HSP70 Heat-Shock Proteins/*analysis/metabolism ; Intercellular Signaling Peptides and Proteins ; Lipid Peroxidation ; Mice ; Mice, Transgenic ; Nerve Tissue Proteins/*analysis/metabolism ; Oxidative Stress ; Phosphopyruvate Hydratase/*analysis/metabolism ; Proteomics ; Spinal Cord/*chemistry/metabolism ; Superoxide Dismutase/genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is an age-related, fatal motor neuron degenerative disease occurring both sporadically (sALS) and heritably (fALS), with inherited cases accounting for approximately 10% of diagnoses. Although multiple mechanisms likely contribute to the pathogenesis of motor neuron injury in ALS, recent advances suggest that oxidative stress may play a significant role in the amplification, and possibly the initiation, of the disease. Lipid peroxidation is one of the several outcomes of oxidative stress. Since the central nervous system (CNS) is enriched with polyunsaturated fatty acids, it is particularly vulnerable to membrane-associated oxidative stress. Peroxidation of cellular membrane lipids or circulating lipoprotein molecules generates highly reactive aldehydes, among which is 4-hydroxy-2-nonenal (HNE). HNE levels are increased in spinal cord motor neurons of ALS patients, indicating that lipid peroxidation is associated with the motor neuron degeneration in ALS. In the present study, we used a parallel proteomic approach to identify HNE-modified proteins in the spinal cord tissue of a model of fALS, G93A-SOD1 transgenic mice, in comparison to the nontransgenic mice. We found three significantly HNE-modified proteins in the spinal cord of G93A-SOD1 transgenic mice: dihydropyrimidinase-related protein 2 (DRP-2), heat-shock protein 70 (Hsp70), and possibly alpha-enolase. These results support the role of oxidative stress as a major mechanism in the pathogenesis of ALS. Structural alteration and activity decline of functional proteins may consistently contribute to the neurodegeneration process in ALS.}, }
@article {pmid15836922, year = {2005}, author = {Patel, YJ and Payne Smith, MD and de Belleroche, J and Latchman, DS}, title = {Hsp27 and Hsp70 administered in combination have a potent protective effect against FALS-associated SOD1-mutant-induced cell death in mammalian neuronal cells.}, journal = {Brain research. Molecular brain research}, volume = {134}, number = {2}, pages = {256-274}, doi = {10.1016/j.molbrainres.2004.10.028}, pmid = {15836922}, issn = {0169-328X}, mesh = {Amino Acid Chloromethyl Ketones/pharmacology ; Amyotrophic Lateral Sclerosis/genetics/pathology/physiopathology/*prevention & control ; Analysis of Variance ; Animals ; Animals, Newborn ; Blotting, Western/methods ; Cell Death/drug effects/genetics ; Cells, Cultured ; Cricetinae ; Culture Media, Serum-Free/pharmacology ; Disease Models, Animal ; Drug Combinations ; Drug Interactions ; Enzyme Inhibitors/pharmacology ; Ganglia, Spinal/*cytology ; Genetic Vectors/physiology ; Green Fluorescent Proteins/metabolism ; HIV-1/physiology ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins/*administration & dosage ; Heat-Shock Proteins/*administration & dosage ; In Situ Nick-End Labeling/methods ; Mutagenesis/physiology ; Mutation ; Neoplasm Proteins/*administration & dosage ; Neurons/*drug effects ; Neuroprotective Agents/pharmacology ; Rats ; Staurosporine/pharmacology ; Superoxide Dismutase/genetics ; Superoxide Dismutase-1 ; Time Factors ; Transfection/methods ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is an adult-onset degenerative disorder characterised by the death of motor neurons in the cortex, brainstem, and spinal cord; resulting in progressive muscle weakness, atrophy, and death from respiratory paralysis, usually within 3-5 years of symptom onset. Approximately 10% of ALS cases are familial (FALS). Mutations in superoxide dismutase-1 (SOD1) cause approximately 20% of FALS cases and there is overwhelming evidence that a toxic gain of function is the cause of the disease. We have previously shown that FALS-associated SOD1 disease mutants enhanced neuronal death in response to a wide range of stimuli tested whereas wt-SOD1 protected against all insults. We demonstrate for the first time that over-expression of either heat shock protein Hsp27 or Hsp70 has a protective effect against SOD1 disease associated mutant-induced cell death. However, over-expression of Hsp27 and Hsp70 together has a greater potent anti-apoptotic effect, than when expressed singly, against the damaging effects of mutant SOD1. Our results indicate that FALS-associated SOD1 disease mutants possess enhanced death-inducing properties and lead to increased apoptosis which can be prevented by either the use of specific caspase inhibitors or Hsp27 and/or Hsp70 over-expression. This potent protective effect of Hsp27 and Hsp70 against the FALS-associated SOD1 disease mutants may be of potential therapeutic importance.}, }
@article {pmid15857390, year = {2005}, author = {Liu, J and Shinobu, LA and Ward, CM and Young, D and Cleveland, DW}, title = {Elevation of the Hsp70 chaperone does not effect toxicity in mouse models of familial amyotrophic lateral sclerosis.}, journal = {Journal of neurochemistry}, volume = {93}, number = {4}, pages = {875-882}, doi = {10.1111/j.1471-4159.2005.03054.x}, pmid = {15857390}, issn = {0022-3042}, support = {R37 NS027036/NS/NINDS NIH HHS/United States ; NS27036/NS/NINDS NIH HHS/United States ; }, mesh = {Age Factors ; Age of Onset ; *Amyotrophic Lateral Sclerosis/genetics/metabolism/physiopathology ; Animals ; Behavior, Animal/physiology ; Blotting, Western/methods ; Body Weight/genetics ; Brain/anatomy & histology/metabolism ; Disease Models, Animal ; Gene Expression Regulation/*physiology ; HSP40 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins/genetics/*metabolism ; HSP90 Heat-Shock Proteins/genetics/metabolism ; Heat-Shock Proteins/genetics/metabolism ; Immunohistochemistry/methods ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/metabolism ; Psychomotor Performance/physiology ; Spinal Cord/cytology ; Superoxide Dismutase/genetics ; Superoxide Dismutase-1 ; }, abstract = {Mutations in copper/zinc superoxide dismutase (SOD1) account for 10-20% of a familial form of amyotrophic lateral sclerosis (ALS). A common feature of SOD1 mutants is abnormal aggregation of the aberrant SOD1 in neurons and glia. We now report that in ALS transgenic mouse models the constitutively expressed heat shock protein 70 (Hsp70) is mislocalized into aggregates together with mutant SOD1 and ubiquitin. Forcing increased synthesis of Hsp70 ameliorates both aggregate formation and toxicity in primary motor neurons in culture. However, chronic increase in an inducible form of Hsp70 to about 10-fold its normal level is shown here not to affect disease course or pathology developed in mice from accumulation of any of three familial ALS causing SOD1 mutants with different underlying biochemical characteristics. Therefore, increasing Hsp70 to a level that is protective in mouse models of acute ischemic insult and selected neurodegenerative disorders is not sufficient to ameliorate mutant SOD1-mediated toxicity.}, }
@article {pmid15872021, year = {2005}, author = {Kirby, J and Halligan, E and Baptista, MJ and Allen, S and Heath, PR and Holden, H and Barber, SC and Loynes, CA and Wood-Allum, CA and Lunec, J and Shaw, PJ}, title = {Mutant SOD1 alters the motor neuronal transcriptome: implications for familial ALS.}, journal = {Brain : a journal of neurology}, volume = {128}, number = {Pt 7}, pages = {1686-1706}, doi = {10.1093/brain/awh503}, pmid = {15872021}, issn = {1460-2156}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/metabolism/pathology ; Antioxidants ; Apoptosis/genetics ; Cell Line ; Cell Survival/genetics ; DNA-Binding Proteins/*genetics ; Female ; Gene Expression Profiling ; *Gene Expression Regulation ; Humans ; Male ; Middle Aged ; Motor Neurons/*metabolism/pathology ; *Mutation ; NF-E2-Related Factor 2 ; Nerve Degeneration/genetics ; Oligonucleotide Array Sequence Analysis ; Response Elements ; Superoxide Dismutase/*genetics/metabolism ; Superoxide Dismutase-1 ; Trans-Activators/*genetics ; }, abstract = {Familial amyotrophic lateral sclerosis (FALS) is caused, in 20% of cases, by mutations in the Cu/Zn superoxide dismutase gene (SOD1). Although motor neuron injury occurs through a toxic gain of function, the precise mechanism(s) remains unclear. Using an established NSC34 cellular model for SOD1-associated FALS, we investigated the effects of mutant SOD1 specifically in cells modelling the vulnerable cell population, the motor neurons, without contamination from non-neuronal cells present in CNS. Using gene expression profiling, 268 transcripts were differentially expressed in the presence of mutant human G93A SOD1. Of these, 197 were decreased, demonstrating that the presence of mutant SOD1 leads to a marked degree of transcriptional repression. Amongst these were a group of antioxidant response element (ARE) genes encoding phase II detoxifying enzymes and antioxidant response proteins (so-called 'programmed cell life' genes), the expression of which is regulated by the transcription factor NRF2. We provide evidence that dysregulation of Nrf2 and the ARE, coupled with reduced pentose phosphate pathway activity and decreased generation of NADPH, represent significant and hitherto unrecognized components of the toxic gain of function of mutant SOD1. Other genes of interest significantly altered in the presence of mutant SOD1 include several previously implicated in neurodegeneration, as well as genes involved in protein degradation, the immune response, cell death/survival and the heat shock response. Preliminary studies on isolated motor neurons from SOD1-associated motor neuron disease cases suggest key genes are also differently expressed in the human disease.}, }
@article {pmid15896810, year = {2005}, author = {Calabrese, V and Lodi, R and Tonon, C and D'Agata, V and Sapienza, M and Scapagnini, G and Mangiameli, A and Pennisi, G and Stella, AM and Butterfield, DA}, title = {Oxidative stress, mitochondrial dysfunction and cellular stress response in Friedreich's ataxia.}, journal = {Journal of the neurological sciences}, volume = {233}, number = {1-2}, pages = {145-162}, doi = {10.1016/j.jns.2005.03.012}, pmid = {15896810}, issn = {0022-510X}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aging/physiology ; Animals ; Antioxidants/therapeutic use ; DNA, Mitochondrial/genetics/metabolism ; Energy Metabolism/physiology ; Friedreich Ataxia/genetics/metabolism/*physiopathology ; Heat-Shock Proteins/metabolism ; Humans ; Mitochondrial Diseases/genetics/metabolism/*physiopathology ; Neurodegenerative Diseases/drug therapy/genetics/metabolism/*physiopathology ; Oxidative Stress/*physiology ; Reactive Nitrogen Species/metabolism ; Reactive Oxygen Species/metabolism ; }, abstract = {There is significant evidence that the pathogenesis of several neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease, Friedreich's ataxia (FRDA), multiple sclerosis and amyotrophic lateral sclerosis, may involve the generation of reactive oxygen species (ROS) and/or reactive nitrogen species (RNS) associated with mitochondrial dysfunction. The mitochondrial genome may play an essential role in the pathogenesis of these diseases, and evidence for mitochondria being a site of damage in neurodegenerative disorders is based in part on observed decreases in the respiratory chain complex activities in Parkinson's, Alzheimer's, and Huntington's disease. Such defects in respiratory complex activities, possibly associated with oxidant/antioxidant imbalance, are thought to underlie defects in energy metabolism and induce cellular degeneration. The precise sequence of events in FRDA pathogenesis is uncertain. The impaired intramitochondrial metabolism with increased free iron levels and a defective mitochondrial respiratory chain, associated with increased free radical generation and oxidative damage, may be considered possible mechanisms that compromise cell viability. Recent evidence suggests that frataxin might detoxify ROS via activation of glutathione peroxidase and elevation of thiols, and in addition, that decreased expression of frataxin protein is associated with FRDA. Many approaches have been undertaken to understand FRDA, but the heterogeneity of the etiologic factors makes it difficult to define the clinically most important factor determining the onset and progression of the disease. However, increasing evidence indicates that factors such as oxidative stress and disturbed protein metabolism and their interaction in a vicious cycle are central to FRDA pathogenesis. Brains of FRDA patients undergo many changes, such as disruption of protein synthesis and degradation, classically associated with the heat shock response, which is one form of stress response. Heat shock proteins are proteins serving as molecular chaperones involved in the protection of cells from various forms of stress. In the central nervous system, heat shock protein (HSP) synthesis is induced not only after hyperthermia, but also following alterations in the intracellular redox environment. The major neurodegenerative diseases, Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Huntington's disease (HD) and FRDA are all associated with the presence of abnormal proteins. Among the various HSPs, HSP32, also known as heme oxygenase I (HO-1), has received considerable attention, as it has been recently demonstrated that HO-1 induction, by generating the vasoactive molecule carbon monoxide and the potent antioxidant bilirubin, could represent a protective system potentially active against brain oxidative injury. Given the broad cytoprotective properties of the heat shock response there is now strong interest in discovering and developing pharmacological agents capable of inducing the heat shock response. This may open up new perspectives in medicine, as molecules inducing this defense mechanism appear to be possible candidates for novel cytoprotective strategies. In particular, manipulation of endogenous cellular defense mechanisms, such as the heat shock response, through nutritional antioxidants, pharmacological compounds or gene transduction, may represent an innovative approach to therapeutic intervention in diseases causing tissue damage, such as neurodegeneration.}, }
@article {pmid15920739, year = {2005}, author = {McLean, JR and Sanelli, TR and Leystra-Lantz, C and He, BP and Strong, MJ}, title = {Temporal profiles of neuronal degeneration, glial proliferation, and cell death in hNFL(+/+) and NFL(-/-) mice.}, journal = {Glia}, volume = {52}, number = {1}, pages = {59-69}, doi = {10.1002/glia.20218}, pmid = {15920739}, issn = {0894-1491}, mesh = {Amyotrophic Lateral Sclerosis/genetics/metabolism/*physiopathology ; Animals ; Astrocytes/metabolism/pathology ; Caspase 3 ; Caspases/metabolism ; Cell Death/physiology ; Cell Proliferation ; Disease Models, Animal ; Gliosis/genetics/metabolism/*physiopathology ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Inclusion Bodies/genetics/metabolism/pathology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mice, Transgenic ; Microglia/metabolism/pathology ; Motor Neurons/metabolism/pathology ; Nerve Degeneration/genetics/metabolism/*physiopathology ; Neurofilament Proteins/*genetics ; Time Factors ; }, abstract = {Neurofilament (NF) aggregate formation within motor neurons is a pathological hallmark of both the sporadic and familial forms of amyotrophic lateral sclerosis (ALS). The relationship between aggregate formation and both microglial and astrocytic proliferation, as well as additional neuropathological features of ALS, is unknown. To examine this, we have used transgenic mice that develop NF aggregates, through either a lack of the low-molecular-weight NF subunit [NFL (-/-)] or the overexpression of human NFL [hNFL (+/+)]. Transgenic and wild-type C57bl/6 mice were examined from 1 month to 18 months of age, and the temporal pattern of motor neuron degeneration, microglial and astrocytic proliferation, and heat shock protein-70 (HSP-70) expression characterized. We observed three overlapping phases in both transgenic mice, including transient aggregate formation, reactive microgliosis, and progressive motor neuron loss. However, only NFL (-/-) mice demonstrated significant astrogliosis and HSP-70 upregulation in both motor neurons and astrocytes. These in vivo models suggest that the development of NF aggregates in motor neurons leads to motor neuron death, but that the interaction between the degenerating motor neurons and the adjacent non-neuronal cells may differ significantly depending on the etiology of the NF aggregate itself.}, }
@article {pmid15996807, year = {2005}, author = {Koh, SH and Roh, H and Lee, SM and Kim, HJ and Kim, M and Lee, KW and Kim, HT and Kim, J and Kim, SH}, title = {Phosphatidylinositol 3-kinase activator reduces motor neuronal cell death induced by G93A or A4V mutant SOD1 gene.}, journal = {Toxicology}, volume = {213}, number = {1-2}, pages = {45-55}, doi = {10.1016/j.tox.2005.05.009}, pmid = {15996807}, issn = {0300-483X}, mesh = {Amyotrophic Lateral Sclerosis/enzymology ; Apoptosis/*physiology ; Blotting, Western ; Caspase 3 ; Caspases/metabolism ; Cell Survival/drug effects ; Chromones/pharmacology ; Enzyme Activation/drug effects ; Glycogen Synthase Kinase 3/metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Morpholines/pharmacology ; Motor Neurons/cytology/drug effects/*enzymology ; Phosphatidylinositol 3-Kinases/*metabolism ; Phosphoinositide-3 Kinase Inhibitors ; Point Mutation ; Poly(ADP-ribose) Polymerases/metabolism ; Protein Kinase Inhibitors/pharmacology ; Protein Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins/metabolism ; Proto-Oncogene Proteins c-akt ; Signal Transduction ; Superoxide Dismutase/genetics/*metabolism ; }, abstract = {The primary pathogenic mechanism of amyotrophic lateral sclerosis (ALS) remains largely unclear. We recently observed that motoneuron cell death mediated by G93A or A4V mutant SOD1, causing familial ALS, was related with decrease of survival signals, such as phosphatidylinositol 3-kinase (PI3-K) and Akt, which play a pivotal role in neuronal survival. Using a G93A or A4V mutant SOD1 transfected VSC4.1 motoneuron cells (G93A or A4V cells, respectively), we presently investigated whether PI3-K activator could reduce mutant SOD1-mediated motoneuron cell death. To investigate the effect of PI3-K activator on viability of G93A and A4V cells, these cells were treated with 10, 50 or 100ng/ml PI3-K activator for 24h and viability and intracellular signals, including Akt, glycogen synthase kinase-3 (GSK-3), heat shock transcription factor-1 (HSTF-1), cytosolic cytochrome c, caspase-3 and poly(ADP-ribose) polymerase (PARP), were compared with those without treatment (control). Compared with non-treated control G93A or A4V cells, the PI3-K activator treatment increased their viability by enhancing the survival signals, including pAkt, pGSK-3, and by inhibiting the death signals, including caspase-3 activation and PARP cleavage. These results suggest that PI3-K activator protects G93A or A4V cells from mutant SOD1-mediated motoneuron cell death by both activating survival signals and inactivating death signals.}, }
@article {pmid16000321, year = {2005}, author = {Wang, J and Xu, G and Li, H and Gonzales, V and Fromholt, D and Karch, C and Copeland, NG and Jenkins, NA and Borchelt, DR}, title = {Somatodendritic accumulation of misfolded SOD1-L126Z in motor neurons mediates degeneration: alphaB-crystallin modulates aggregation.}, journal = {Human molecular genetics}, volume = {14}, number = {16}, pages = {2335-2347}, doi = {10.1093/hmg/ddi236}, pmid = {16000321}, issn = {0964-6906}, support = {R01 NS 047225/NS/NINDS NIH HHS/United States ; R01 NS 37225/NS/NINDS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/*etiology/pathology ; Animals ; Astrocytes/metabolism/ultrastructure ; Dendrites/*enzymology ; Disease Models, Animal ; Female ; Humans ; Male ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/*enzymology/pathology ; Oligodendroglia/metabolism/ultrastructure ; *Protein Folding ; Sequence Deletion ; Spinal Cord/*enzymology/metabolism/pathology ; Superoxide Dismutase/chemistry/*metabolism ; Superoxide Dismutase-1 ; Up-Regulation ; alpha-Crystallin B Chain/*metabolism ; }, abstract = {Mice expressing variants of superoxide dismutase-1 (SOD1) encoding C-terminal truncation mutations linked to familial amyotrophic lateral sclerosis (FALS) have begun to define the role of misfolding and aggregation in the pathogenesis of disease. Here, we examine transgenic mice expressing SOD1-L126Z (Z = stop-truncation of last 28 amino acids), finding that detergent-insoluble mutant protein specifically accumulates in somatodendritic compartments. Soluble forms of the SOD1-L126Z were virtually undetectable in spinal cord at any age and the levels of accumulated protein directly correlated with disease symptoms. Neither soluble nor insoluble forms of SOD1-L126Z were transported to distal axons. In vitro, small heat shock protein (Hsp) alphaB-crystallin suppressed the in vitro aggregation of SOD1-L126Z. In vivo, alphaB-crystallin immunoreactivity was most abundant in oligodendrocytes and up-regulated in astrocytes of symptomatic mice; neither of these cell-types accumulated mutant SOD1 immunoreactivity. These results suggest that damage to motor neuron cell bodies and dendrites within the spinal cord can be sufficient to induce motor neuron disease and that the activities of chaperones may modulate the cellular specificity of mutant SOD1 accumulation.}, }
@article {pmid16042542, year = {2005}, author = {Kalmar, B and Kieran, D and Greensmith, L}, title = {Molecular chaperones as therapeutic targets in amyotrophic lateral sclerosis.}, journal = {Biochemical Society transactions}, volume = {33}, number = {Pt 4}, pages = {551-552}, doi = {10.1042/BST0330551}, pmid = {16042542}, issn = {0300-5127}, mesh = {Disease Progression ; Heat-Shock Proteins/*physiology ; Humans ; Molecular Chaperones/*physiology ; Motor Neuron Disease/*therapy ; Motor Neurons/*pathology ; Peripheral Nervous System Diseases/pathology ; }, abstract = {Neurodegenerative diseases are characterized by a number of common hallmarks, such as the presence of intracellular aggregates and activation of the apoptotic cell-death pathway. Intracellular chaperones, responsible for protein integrity and structural repair, may play a crucial role in the progression of a disease. In this paper, we aim to summarize our understanding of the role and potential of a particular family of chaperones, the heat-shock proteins, in neurodegeneration, by focusing our discussion on models of motoneuron death.}, }
@article {pmid16045483, year = {2005}, author = {Koh, SH and Lee, YB and Kim, KS and Kim, HJ and Kim, M and Lee, YJ and Kim, J and Lee, KW and Kim, SH}, title = {Role of GSK-3beta activity in motor neuronal cell death induced by G93A or A4V mutant hSOD1 gene.}, journal = {The European journal of neuroscience}, volume = {22}, number = {2}, pages = {301-309}, doi = {10.1111/j.1460-9568.2005.04191.x}, pmid = {16045483}, issn = {0953-816X}, mesh = {Alanine/genetics ; Animals ; Apoptosis/drug effects ; Blotting, Western/methods ; Caspase 3 ; Caspases/metabolism ; Cell Death/drug effects/genetics ; Cell Differentiation ; Cell Line, Tumor ; Cell Survival/drug effects ; Collagen Type XI/metabolism ; Cytochromes c/metabolism ; Enzyme Inhibitors/pharmacology ; Glycogen Synthase Kinase 3/antagonists & inhibitors/*physiology ; Glycogen Synthase Kinase 3 beta ; Heat-Shock Proteins/metabolism ; Humans ; Hybrid Cells ; Indoles/metabolism ; Motor Neurons/drug effects/*physiology ; Mutagenesis/physiology ; Neuroblastoma ; Phosphatidylinositol 3-Kinases/metabolism ; Point Mutation/*genetics ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; Protein Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins/metabolism ; Proto-Oncogene Proteins c-akt ; Superoxide Dismutase/*genetics/physiology ; Tetrazolium Salts ; Thiazoles ; Time Factors ; Transfection/methods ; Trypan Blue ; Valine/genetics ; }, abstract = {Point mutations such as G93A and A4V in the human Cu/Zn-superoxide dismutase gene (hSOD1) cause familial amyotrophic lateral sclerosis (fALS). In spite of several theories to explain the pathogenic mechanisms, the mechanism remains largely unclear. Increased activity of glycogen synthase kinase-3 (GSK-3) has recently been emphasized as an important pathogenic mechanism of neurodegenerative diseases, including Alzheimer's disease and ALS. To investigate the effects of G93A or A4V mutations on the phosphatidylinositol-3-kinase (PI3-K)/Akt and GSK-3 pathway as well as the caspase-3 pathway, VSC4.1 motoneuron cells were transfected with G93A- or A4V-mutant types of hSOD1 (G93A and A4V cells, respectively) and, 24 h after neuronal differentiation, their viability and intracellular signals, including PI3-K/Akt, GSK-3, heat shock transcription factor-1 (HSTF-1), cytochrome c, caspase-3 and poly(ADP-ribose) polymerase (PARP), were compared with those of wild type (wild cells). Furthermore, to elucidate the role of the GSK-3beta-mediated cell death mechanism, alterations of viability and intracellular signals in those mutant motoneurons were investigated after treating the cells with GSK-3beta inhibitor. Compared with wild cells, viability was greatly reduced in the G93A and A4V cells. However, the treatment of G93A and A4V cells with GSK-3beta inhibitor increased their viability by activating HSTF-1 and by reducing cytochrome c release, caspase-3 activation and PARP cleavage. However, the treatment did not affect the expression of PI3-K/Akt and GSK-3beta. These results suggest that the G93A or A4V mutations inhibit PI3-K/Akt and activate GSK-3beta and caspase-3, thus becoming vulnerable to oxidative stress, and that the GSK-3beta-mediated cell death mechanism is important in G93A and A4V cell death.}, }
@article {pmid16048838, year = {2005}, author = {Meriin, AB and Sherman, MY}, title = {Role of molecular chaperones in neurodegenerative disorders.}, journal = {International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group}, volume = {21}, number = {5}, pages = {403-419}, doi = {10.1080/02656730500041871}, pmid = {16048838}, issn = {0265-6736}, mesh = {Apoptosis/physiology ; Cellular Senescence/physiology ; Heat-Shock Proteins/physiology ; Humans ; Inclusion Bodies/metabolism ; Models, Biological ; Molecular Chaperones/chemistry/metabolism/*physiology ; Neurodegenerative Diseases/metabolism/*physiopathology/therapy ; Protein Conformation ; Protein Folding ; Ubiquitin-Protein Ligases/physiology ; }, abstract = {Many major neurodegenerative diseases, including Amyotrophic Lateral Sclerosis, Alzheimer's disease, Parkinson's disease, Huntington Disease and other polyglutamine expansion disorders, are associated with degeneration and death of specific neuronal populations due to accumulation of certain abnormal polypeptides. These misfolded species aggregate and form inclusion bodies and their neurotoxicity is associated with the aggregation. To handle a build-up of abnormal proteins cells employ a complicated machinery of molecular chaperones and various proteolytic systems. Chaperones facilitate refolding or degradation of misfolded polypeptides, prevent protein aggregation and play a role in formation of aggresome, a centrosome-associated body to which small cytoplasmic aggregates are transported. The ubiquitin-proteasome proteolytic system is critical for reducing the levels of soluble abnormal proteins, while autophagy plays the major role in clearing of cells from protein aggregates. Accumulation of the aggregation prone proteins activates signal transduction pathways that control cell death, including JNK pathway that controls viability of a cell in various models of Parkinson's and Huntington's diseases. The major chaperone Hsp72 can interfere with this signalling pathway, thus promoting survival. A very important consequence of a build-up and aggregation of misfolded proteins is impairment of the ubiquitin-proteasome degradation system and suppression of the heat shock response. Such an inhibition of the major cell defense systems may play a critical role in neurodegeneration. Here, it is suggested that these changes may reflect a senescence-like programme initiated by the aggregated abnormal polypeptides. Pathways that control the fate of misfolded proteins, for example molecular chaperones or proteolytic systems, may become interesting novel targets for therapy of neurodegenerative disorders.}, }
@article {pmid16184763, year = {2005}, author = {Batulan, Z and Nalbantoglu, J and Durham, HD}, title = {Nonsteroidal anti-inflammatory drugs differentially affect the heat shock response in cultured spinal cord cells.}, journal = {Cell stress & chaperones}, volume = {10}, number = {3}, pages = {185-196}, pmid = {16184763}, issn = {1355-8145}, mesh = {Amyotrophic Lateral Sclerosis/drug therapy ; Animals ; Anti-Inflammatory Agents, Non-Steroidal/*pharmacology ; Cell Survival ; Cells, Cultured ; Cyclooxygenase Inhibitors/*pharmacology ; Dose-Response Relationship, Drug ; Glial Fibrillary Acidic Protein/analysis ; HSP70 Heat-Shock Proteins/biosynthesis/metabolism ; Heat-Shock Response/*drug effects ; Mice ; Motor Neurons/drug effects/metabolism ; Mutation ; Neurofilament Proteins/analysis ; Neuroglia/*drug effects/metabolism ; Niflumic Acid/*pharmacology ; Plasmids ; Sodium Salicylate/*pharmacology ; Spinal Cord/*drug effects/embryology ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1 ; Temperature ; }, abstract = {Nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to amplify the heat shock response in cell lines by increasing the binding of heat shock transcription factor-1 to heat shock elements within heat shock gene promoters. Because overexpression of the inducible heat shock protein 70 (Hsp70) was neuroprotective in a culture model of motor neuron disease, this study investigated whether NSAIDs induce Hsp70 and confer cytoprotection in motor neurons of dissociated spinal cord cultures exposed to various stresses. Two NSAIDs, sodium salicylate and niflumic acid, lowered the temperature threshold for induction of Hsp70 in glia but failed to do so in motor neurons. At concentrations that increased Hsp70 in heat shocked glial cells, sodium salicylate failed to delay death of motor neurons exposed to hyperthermia, paraquat-mediated oxidative stress, and glutamate excitotoxicity. Neither sodium salicylate nor the cyclooxygenase-2 inhibitor, niflumic acid, protected motor neurons from the toxicity of mutated Cu/Zn-superoxide dismutase (SOD-1) linked to a familial form of the motor neuron disease, amyotrophic lateral sclerosis. Thus, treatment with 2 types of NSAIDs failed to overcome the high threshold for the activation of heat shock response in motor neurons.}, }
@article {pmid16260738, year = {2005}, author = {Katsuno, M and Sang, C and Adachi, H and Minamiyama, M and Waza, M and Tanaka, F and Doyu, M and Sobue, G}, title = {Pharmacological induction of heat-shock proteins alleviates polyglutamine-mediated motor neuron disease.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {102}, number = {46}, pages = {16801-16806}, pmid = {16260738}, issn = {0027-8424}, mesh = {Animals ; Cell Line, Tumor ; Cell Nucleus/drug effects/metabolism ; Diterpenes/*pharmacology ; Heat-Shock Proteins/*biosynthesis/genetics ; Humans ; Mice ; Motor Neuron Disease/chemically induced/*metabolism/physiopathology ; Peptides/*toxicity ; Receptors, Androgen/metabolism ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is an adult-onset motor neuron disease caused by the expansion of a trinucleotide CAG repeat encoding the polyglutamine tract in the first exon of the androgen receptor gene (AR). The pathogenic, polyglutamine-expanded AR protein accumulates in the cell nucleus in a ligand-dependent manner and inhibits transcription by interfering with transcriptional factors and coactivators. Heat-shock proteins (HSPs) are stress-induced chaperones that facilitate the refolding and, thus, the degradation of abnormal proteins. Geranylgeranylacetone (GGA), a nontoxic antiulcer drug, has been shown to potently induce HSP expression in various tissues, including the central nervous system. In a cell model of SBMA, GGA increased the levels of Hsp70, Hsp90, and Hsp105 and inhibited cell death and the accumulation of pathogenic AR. Oral administration of GGA also up-regulated the expression of HSPs in the central nervous system of SBMA-transgenic mice and suppressed nuclear accumulation of the pathogenic AR protein, resulting in amelioration of polyglutamine-dependent neuromuscular phenotypes. These observations suggest that, although a high dose appears to be needed for clinical effects, oral GGA administration is a safe and promising therapeutic candidate for polyglutamine-mediated neurodegenerative diseases, including SBMA.}, }
@article {pmid16280684, year = {2005}, author = {Nirmalananthan, N and Greensmith, L}, title = {Amyotrophic lateral sclerosis: recent advances and future therapies.}, journal = {Current opinion in neurology}, volume = {18}, number = {6}, pages = {712-719}, doi = {10.1097/01.wco.0000187248.21103.c5}, pmid = {16280684}, issn = {1350-7540}, support = {G84/6699/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics/metabolism/*therapy ; Animals ; Biomedical Research ; Docosahexaenoic Acids/classification/*therapeutic use ; Drug Evaluation, Preclinical ; Forecasting ; Humans ; }, abstract = {PURPOSE OF REVIEW: Amyotrophic lateral sclerosis is a rare but fatal motoneuron disorder. Despite intensive research riluzole remains the only available therapy, with only marginal effects on survival. Here we review some of the recent advances in the search for a disease-modifying therapy for amyotrophic lateral sclerosis.<br><br>RECENT FINDINGS: A number of established agents have recently been re-investigated for their potential as neuroprotective agents, including beta-lactam antibiotics and minocycline. Progress has also been made in exploiting growth factors for the treatment of amyotrophic lateral sclerosis, partly due to advances in developing effective delivery systems to the central nervous system. A number of new therapies have also been identified, including a novel class of compounds, heat-shock protein co-inducers, which upregulate cell stress responses thereby mediating neuroprotection. Non-drug-based therapies are also under development, with progress in gene-silencing and stem cell therapies.<br><br>SUMMARY: In the past few years, significant advances have been made in both our understanding of amyotrophic lateral sclerosis pathogenesis and the development of new therapeutic approaches. However, caution must be exercised in view of the long-standing failure to successfully transfer therapeutic compounds to the clinic. A deeper awareness in the research community of the need for clinically relevant preclinical studies, coupled with a better understanding of the issues surrounding clinical trial design for amyotrophic lateral sclerosis, offers hope that the growing list of validated preclinical therapeutics can finally yield an effective disease-modifying treatment.}, }
@article {pmid16364298, year = {2006}, author = {Tovar-y-Romo, LB and Tapia, R}, title = {Cerebral neurons of transgenic ALS mice are vulnerable to glutamate release stimulation but not to increased extracellular glutamate due to transport blockade.}, journal = {Experimental neurology}, volume = {199}, number = {2}, pages = {281-290}, doi = {10.1016/j.expneurol.2005.11.005}, pmid = {16364298}, issn = {0014-4886}, mesh = {4-Aminopyridine/pharmacology ; Amyotrophic Lateral Sclerosis/*pathology ; Animals ; Biological Transport/drug effects ; Dicarboxylic Acids/pharmacology ; Disease Models, Animal ; Dizocilpine Maleate/pharmacology ; Excitatory Amino Acid Agonists/pharmacology ; Excitatory Amino Acid Antagonists/pharmacology ; Extracellular Space/drug effects/*metabolism ; Glutamic Acid/*metabolism ; HSP72 Heat-Shock Proteins/metabolism ; Humans ; Mice ; Mice, Transgenic ; Microdialysis/methods ; Neurons/drug effects/*metabolism ; Neurotransmitter Uptake Inhibitors/pharmacology ; Pyrrolidines/pharmacology ; Superoxide Dismutase/genetics ; }, abstract = {Mechanisms of motor neuron loss in amyotrophic lateral sclerosis (ALS) are unknown, but it has been postulated that excitotoxicity due to excessive glutamatergic neurotransmission by decreased efficiency of glutamate transport may be involved in both familial (FALS) and sporadic ALS. Using microdialysis in vivo, we tested the effects of the glutamate transport inhibitor L-trans-pyrrolidine-2,4-dicarboxylate (PDC) and of 4-aminopyridine (4-AP), which stimulates glutamate release from nerve endings, in the hippocampus and motor cortex of wild type (WT) and transgenic SOD1/G93A mice, an established model of FALS. Perfusion of 4-AP induced convulsions, expression of the inducible stress-marker heat-shock protein 70 (HSP70) and hippocampal neuronal loss. These effects were similar in both WT and G93A mice, and, in both groups, they were prevented by the previous systemic administration of the NMDA receptor antagonist MK-801. In contrast, perfusion of PDC resulted in a large and long-lasting (2 h) increase of extracellular glutamate, but no convulsions, neuronal damage or HSP70 expression were observed in either the WT or the G93A mice. Our results demonstrate that SOD1 G93A mutation does not enhance the vulnerability to endogenous glutamate-mediated excitotoxicity in brain, neither by blocking glutamate transport nor by stimulating its release. Therefore, these data do not support the possibility that glutamate transport deficiency may be an important factor of brain neuronal degeneration in familial ALS.}, }
@article {pmid16741751, year = {2006}, author = {Waza, M and Adachi, H and Katsuno, M and Minamiyama, M and Tanaka, F and Doyu, M and Sobue, G}, title = {Modulation of Hsp90 function in neurodegenerative disorders: a molecular-targeted therapy against disease-causing protein.}, journal = {Journal of molecular medicine (Berlin, Germany)}, volume = {84}, number = {8}, pages = {635-646}, pmid = {16741751}, issn = {0946-2716}, mesh = {Age Factors ; Animals ; Benzoquinones/pharmacology/therapeutic use ; HSP90 Heat-Shock Proteins/antagonists &amp; inhibitors/genetics/*physiology ; Lactams, Macrocyclic/pharmacology/therapeutic use ; Mice ; Motor Neurons/drug effects/pathology ; Muscular Disorders, Atrophic/drug therapy/metabolism/pathology ; Mutation ; Neurodegenerative Diseases/drug therapy/*metabolism/pathology ; Peptides/genetics ; Proteasome Endopeptidase Complex/metabolism ; Receptors, Androgen/genetics/metabolism ; }, abstract = {Abnormal accumulation of disease-causing protein is a commonly observed characteristic in chronic neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and polyglutamine (polyQ) diseases. A therapeutic approach that could selectively eliminate would be a promising remedy for neurodegenerative disorders. Spinal and bulbar muscular atrophy (SBMA), one of the polyQ diseases, is a late-onset motor neuron disease characterized by proximal muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. The pathogenic gene product is polyQ-expanded androgen receptor (AR), which belongs to the heat shock protein (Hsp) 90 client protein family. 17-Allylamino-17-demethoxygeldanamycin (17-AAG), a novel Hsp90 inhibitor, is a new derivative of geldanamycin that shares its important biological activities but shows less toxicity. 17-AAG is now in phase II clinical trials as a potential anti-cancer agent because of its ability to selectively degrade several oncoproteins. We have recently demonstrated the efficacy and safety of 17-AAG in a mouse model of SBMA. The administration of 17-AAG significantly ameliorated polyQ-mediated motor neuron degeneration by reducing the total amount of mutant AR. 17-AAG accomplished the preferential reduction of mutant AR mainly through Hsp90 chaperone complex formation and subsequent proteasome-dependent degradation. 17-AAG induced Hsp70 and Hsp40 in vivo as previously reported; however, its ability to induce HSPs was limited, suggesting that the HSP induction might support the degradation of mutant protein. The ability of 17-AAG to preferentially degrade mutant protein would be directly applicable to SBMA and other neurodegenerative diseases in which the disease-causing proteins also belong to the Hsp90 client protein family. Our proposed therapeutic approach, modulation of Hsp90 function by 17-AAG treatment, has emerged as a candidate for molecular-targeted therapies for neurodegenerative diseases. This review will consider our research findings and discuss the possibility of a clinical application of 17-AAG to SBMA and other neurodegenerative diseases.}, }
@article {pmid16806187, year = {2006}, author = {Krishnan, J and Lemmens, R and Robberecht, W and Van Den Bosch, L}, title = {Role of heat shock response and Hsp27 in mutant SOD1-dependent cell death.}, journal = {Experimental neurology}, volume = {200}, number = {2}, pages = {301-310}, doi = {10.1016/j.expneurol.2006.02.135}, pmid = {16806187}, issn = {0014-4886}, mesh = {Animals ; Antirheumatic Agents/pharmacology ; Blotting, Western/methods ; Cell Death/drug effects/genetics ; Cell Line, Tumor ; Cyclosporine/pharmacology ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/*physiology ; Heat-Shock Response/drug effects/*physiology ; Humans ; Immunohistochemistry/methods ; Mice ; Mutagenesis/physiology ; Mutant Proteins/*genetics ; Neuroblastoma ; RNA, Messenger/metabolism ; RNA, Small Interfering/pharmacology ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Superoxide Dismutase/*genetics ; Superoxide Dismutase-1 ; Time Factors ; Transfection/methods ; }, abstract = {The fatal neurodegenerative disorder amyotrophic lateral sclerosis (ALS) is characterized by selective loss of motor neurons and mutations in the copper-zinc superoxide dismutase (SOD1) enzyme underlie one form of familial ALS. The pathogenic mechanism of these mutations is elusive but is thought to involve oxidative stress and protein aggregation. These two phenomena are known to induce heat shock proteins (Hsps) which protect stressed cells through their chaperoning and anti-apoptotic activity. In order to investigate the role of Hsp27 in mutant SOD1-dependent cell death, we used mutant and wild type SOD1 overexpressing N2a mouse neuroblastoma cells. Mutant SOD1-dependent cell death could be induced by heat shock, and by treating the cells with cyclosporine A or lactacystin. Transfection with an Hsp27 expression construct did not protect the N2a cells against mutant SOD1-dependent cell death. However, pre-conditioning N2a cells with a mild heat shock was accompanied by a significant upregulation of Hsp27 in the mutant SOD1 cells, and protected these cells against subsequent cell death induced by a more severe heat shock. Selective inhibition of the Hsp27 upregulation, through the use of Hsp27 siRNA, did not attenuate the protective effect of this treatment. These results show that activation of the heat shock response protects cells against mutant SOD1-dependent cell death, but that Hsp27 is not an essential component of the stress response leading to protection.}, }
@article {pmid16909005, year = {2005}, author = {Kiaei, M and Kipiani, K and Petri, S and Chen, J and Calingasan, NY and Beal, MF}, title = {Celastrol blocks neuronal cell death and extends life in transgenic mouse model of amyotrophic lateral sclerosis.}, journal = {Neuro-degenerative diseases}, volume = {2}, number = {5}, pages = {246-254}, doi = {10.1159/000090364}, pmid = {16909005}, issn = {1660-2854}, mesh = {Amyotrophic Lateral Sclerosis/drug therapy/genetics/*pathology ; Animals ; Astrocytes/drug effects/metabolism ; CD40 Antigens/metabolism ; Cell Count ; Cell Death/drug effects ; Female ; Fluorescent Antibody Technique, Indirect ; Glial Fibrillary Acidic Protein/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Longevity ; Mice ; Mice, Transgenic ; Motor Neurons/drug effects/metabolism ; Neuroglia/drug effects ; Neurons/*drug effects/metabolism ; Neuroprotective Agents/*pharmacology/therapeutic use ; Pentacyclic Triterpenes ; Postural Balance/drug effects ; Psychomotor Performance ; Superoxide Dismutase/genetics ; Superoxide Dismutase-1 ; Survival Analysis ; Triterpenes/*pharmacology/therapeutic use ; Tumor Necrosis Factor-alpha/metabolism ; Up-Regulation ; }, abstract = {There is substantial evidence that both inflammation and oxidative damage contribute to the pathogenesis of motor neuron degeneration in the G93A SOD1 transgenic mouse model of amyotrophic lateral sclerosis (ALS). Celastrol is a natural product from Southern China, which exerts potent anti-inflammatory and antioxidative effects. It also acts potently to increase expression of heat shock proteins including HSP70. We administered it in the diet to G93A SOD1 mice starting at 30 days of age. Celastrol treatment significantly improved weight loss, motor performance and delayed the onset of ALS. Survival of celastrol-treated G93A mice increased by 9.4% and 13% for 2 mg/kg/day and 8 mg/kg/day doses, respectively. Cell counts of lumbar spinal cord neurons confirmed a protective effect, i.e. 30% increase in neuronal number in the lumbar spinal cords of celastrol-treated animals. Celastrol treatment reduced TNF-alpha, iNOS, CD40, and GFAP immunoreactivity in the lumbar spinal cord sections of celastrol-treated G93A mice compared to untreated G93A mice. TNF-alpha immunoreactivity co-localized with SMI-32 (neuronal marker) and GFAP (astrocyte marker). HSP70 immunoreactivity was increased in lumbar spinal cord neurons of celastrol-treated G93A mice. Celastrol has been widely used in treating inflammatory diseases in man, and is well tolerated; therefore, it may be a promising therapeutic candidate for the treatment of human ALS.}, }
@article {pmid16950627, year = {2006}, author = {Batulan, Z and Taylor, DM and Aarons, RJ and Minotti, S and Doroudchi, MM and Nalbantoglu, J and Durham, HD}, title = {Induction of multiple heat shock proteins and neuroprotection in a primary culture model of familial amyotrophic lateral sclerosis.}, journal = {Neurobiology of disease}, volume = {24}, number = {2}, pages = {213-225}, doi = {10.1016/j.nbd.2006.06.017}, pmid = {16950627}, issn = {0969-9961}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism/physiopathology ; Animals ; Benzoquinones/pharmacology ; Cells, Cultured ; Cytoprotection/drug effects/*genetics ; DNA-Binding Proteins/*genetics ; Dose-Response Relationship, Drug ; Enzyme Inhibitors/pharmacology ; Gene Expression Regulation, Enzymologic/drug effects/genetics ; Genetic Vectors ; HSP40 Heat-Shock Proteins/antagonists &amp; inhibitors/metabolism ; HSP70 Heat-Shock Proteins/antagonists &amp; inhibitors/metabolism ; HSP90 Heat-Shock Proteins/antagonists &amp; inhibitors/genetics/metabolism ; Heat Shock Transcription Factors ; Heat-Shock Proteins/antagonists &amp; inhibitors/genetics/*metabolism ; Humans ; Lactams, Macrocyclic/pharmacology ; Macrolides/pharmacology ; Mice ; Motor Neurons/drug effects/*metabolism/pathology ; Neuroprotective Agents/chemistry/*pharmacology ; Pyrrolidines/pharmacology ; Superoxide Dismutase/antagonists &amp; inhibitors/toxicity ; Thiocarbamates/pharmacology ; Transcription Factors/*genetics ; Transfection ; Up-Regulation/drug effects/genetics ; }, abstract = {High threshold for stress-induced activation of the heat shock transcription factor, Hsf1, may contribute to vulnerability of motor neurons to disease and limit efficacy of agents promoting expression of neuroprotective heat shock proteins (Hsps) through this transcription factor. Plasmid encoding a constitutively active form of Hsf1, Hsf1act, and chemicals shown to activate Hsf1 in other cells were investigated in a primary culture model of familial amyotrophic lateral sclerosis. Hsf1act and the Hsp90 inhibitor, geldanamycin, induced high expression of multiple Hsps in cultured motor neurons and conferred dramatic neuroprotection against SOD1G93A in comparison to Hsp70 or Hsp25 alone. Two other Hsp90 inhibitors, 17-allylamino-17-demethoxygeldanamycin (17-AAG) and radicicol, and pyrrolidine dithiocarbamate induced robust expression of Hsp70 and Hsp40 in motor neurons, but at cytotoxic concentrations. 17-AAG, which penetrates the blood-brain barrier, has exhibited a higher therapeutic index than geldanamycin, but this may not be the case when activation of Hsf1 in neurons is targeted.}, }
@article {pmid17037196, year = {2006}, author = {Shan, XY and Li, B and Zhang, JR}, title = {[Produce of marker-free transgenic tobacco plants by FLP/frt recombination system].}, journal = {Sheng wu gong cheng xue bao = Chinese journal of biotechnology}, volume = {22}, number = {5}, pages = {744-750}, doi = {10.1016/s1872-2075(06)60054-x}, pmid = {17037196}, issn = {1000-3061}, mesh = {Base Sequence ; DNA Nucleotidyltransferases/*metabolism ; Molecular Sequence Data ; Plants, Genetically Modified/*genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; *Recombination, Genetic ; Nicotiana/*genetics ; }, abstract = {Selectable marker genes that usually encode antibiotic or herbicide resistances are widely used for the selection of the transgenic plants, but they become unnecessary and undesirable after transformation selection. An important strategy to improve the transgenic plants' biosafety is to eliminate the marker genes after successful selection. In the FLP/frt site-specific system of the 2 microm plasmid of Saccharomyces cerevisiae, the FLP enzyme efficiently catalyzes recombination between two directly repeated FLP recombination target (frt) sites, eliminating the sequence between them. By controlled expression of the FLP recombinase and specific allocation of the frt sites within transgenic constructs, the system can be applied to eliminate the marker genes after selection. Through a series of procedures, the plant FLP/frt site-specific recombination system was constructed, which included the frt containing vector pCAMBIA1300-betA-frt-als-frt and the FLP expression vector pCAMBIA1300-hsp-FLP-hpt. The FLP recombinase gene was introduced into transgenic (betA-frt-als-frt) tobacco plants by re-transformation. In re-transgenic plants, after heat shock treatment, the marker gene als flanked by two identical orientation frt sites could be excised by the inducible expression of FLP recombinase under the control of hsp promoter. Excision of the als gene was found in 41% re-transgenic tobacco plants, which indicated that this systerm could make a great contribution to obtain the marker free transgenic plants.}, }
@article {pmid17114821, year = {2007}, author = {Mattson, MP and Cutler, RG and Camandola, S}, title = {Energy intake and amyotrophic lateral sclerosis.}, journal = {Neuromolecular medicine}, volume = {9}, number = {1}, pages = {17-20}, pmid = {17114821}, issn = {1559-1174}, support = {//Intramural NIH HHS/United States ; }, mesh = {Aging ; Amyotrophic Lateral Sclerosis/*metabolism/mortality/physiopathology ; Animals ; Caloric Restriction ; Dietary Fats/administration &amp; dosage ; Dietary Sucrose/administration &amp; dosage ; *Energy Intake ; Male ; Mice ; Motor Neurons/metabolism ; Mutation ; Superoxide Dismutase/genetics ; Survival Rate ; }, abstract = {Roy Walford, a physician and scientist who pioneered research on the anti-aging effects of caloric restriction and subjected himself to a low-energy diet, recently died from amyotrophic lateral sclerosis (ALS). Information from his case, epidemiological findings, and recent controlled studies in mouse models of ALS suggest that low-energy diets might render motor neurons vulnerable to degeneration, whereas high-energy diets are ameliorative. This contrasts with the effects of low-energy diets on various neuronal populations in the brain that respond adaptively, activating pathways that promote plasticity and resistance to disease. One reason that motor neurons might be selectively vulnerable to low-energy diets is that they are unable to engage neuroprotective responses to energetic stress response involving the protein chaperones, such as, heat-shock protein-70.}, }
@article {pmid17191135, year = {2007}, author = {Calabrese, V and Guagliano, E and Sapienza, M and Panebianco, M and Calafato, S and Puleo, E and Pennisi, G and Mancuso, C and Butterfield, DA and Stella, AG}, title = {Redox regulation of cellular stress response in aging and neurodegenerative disorders: role of vitagenes.}, journal = {Neurochemical research}, volume = {32}, number = {4-5}, pages = {757-773}, pmid = {17191135}, issn = {0364-3190}, support = {AG 05119/AG/NIA NIH HHS/United States ; AG 10836/AG/NIA NIH HHS/United States ; }, mesh = {Aging/*metabolism ; Alzheimer Disease/genetics/metabolism ; Animals ; Gene Expression ; Heme Oxygenase (Decyclizing)/metabolism ; Humans ; Longevity/*genetics ; Neurodegenerative Diseases/genetics/*physiopathology ; Oxidation-Reduction ; Oxidative Stress/*genetics/*physiology ; Reactive Oxygen Species/metabolism ; Thioredoxins/metabolism ; }, abstract = {Reduced expression and/or activity of antioxidant proteins lead to oxidative stress, accelerated aging and neurodegeneration. However, while excess reactive oxygen species (ROS) are toxic, regulated ROS play an important role in cell signaling. Perturbation of redox status, mutations favoring protein misfolding, altered glyc(osyl)ation, overloading of the product of polyunsaturated fatty acid peroxidation (hydroxynonenals, HNE) or cholesterol oxidation, can disrupt redox homeostasis. Collectively or individually these effects may impose stress and lead to accumulation of unfolded or misfolded proteins in brain cells. Alzheimer's (AD), Parkinson's and Huntington's disease, amyotrophic lateral sclerosis and Friedreich's ataxia are major neurological disorders associated with production of abnormally aggregated proteins and, as such, belong to the so-called "protein conformational diseases". The pathogenic aggregation of proteins in non-native conformation is generally associated with metabolic derangements and excessive production of ROS. The "unfolded protein response" has evolved to prevent accumulation of unfolded or misfolded proteins. Recent discoveries of the mechanisms of cellular stress signaling have led to new insights into the diverse processes that are regulated by cellular stress responses. The brain detects and overcomes oxidative stress by a complex network of "longevity assurance processes" integrated to the expression of genes termed vitagenes. Heat-shock proteins are highly conserved and facilitate correct protein folding. Heme oxygenase-1, an inducible and redox-regulated enzyme, has having an important role in cellular antioxidant defense. An emerging concept is neuroprotection afforded by heme oxygenase by its heme degrading activity and tissue-specific antioxidant effects, due to its products carbon monoxide and biliverdin, which is then reduced by biliverdin reductase in bilirubin. There is increasing interest in dietary compounds that can inhibit, retard or reverse the steps leading to neurodegeneration in AD. Specifically any dietary components that inhibit inappropriate inflammation, AbetaP oligomerization and consequent increased apoptosis are of particular interest, with respect to a chronic inflammatory response, brain injury and beta-amyloid associated pathology. Curcumin and ferulic acid, the first from the curry spice turmeric and the second a major constituent of fruit and vegetables, are candidates in this regard. Not only do these compounds serve as antioxidants but, in addition, they are strong inducers of the heat-shock response. Food supplementation with curcumin and ferulic acid are therefore being considered as a novel nutritional approach to reduce oxidative damage and amyloid pathology in AD. We review here some of the emerging concepts of pathways to neurodegeneration and how these may be overcome by a nutritional approach.}, }
@article {pmid17274531, year = {2006}, author = {Calabrese, V and Guagliano, E and Sapienza, M and Mancuso, C and Butterfield, DA and Stella, AM}, title = {Redox regulation of cellular stress response in neurodegenerative disorders.}, journal = {The Italian journal of biochemistry}, volume = {55}, number = {3-4}, pages = {263-282}, pmid = {17274531}, issn = {0021-2938}, mesh = {Animals ; Antioxidants/physiology ; Female ; Free Radicals/metabolism ; Heme Oxygenase (Decyclizing)/physiology ; Humans ; Longevity/physiology ; Male ; Neurodegenerative Diseases/*physiopathology ; Nitric Oxide/physiology ; Nitric Oxide Synthase Type II/metabolism ; Oxidation-Reduction ; Oxidoreductases Acting on CH-CH Group Donors/physiology ; Reactive Nitrogen Species/physiology ; Reactive Oxygen Species/*metabolism ; Signal Transduction ; Stress, Physiological/*physiopathology ; Thioredoxin-Disulfide Reductase/physiology ; Thioredoxins/metabolism ; }, abstract = {There is increasing evidence that reactive oxygen species (ROS) are not only toxic but play an important role in cellular signaling and in the regulation of gene expression. A number of biochemical and physiologic stimuli, such as perturbation in redox status, expression of misfolded proteins, altered glyc(osyl)ation and glucose deprivation, overloading of products of polyunsaturated fatty acid peroxidation (Hydroxynonenals, HNE) or cholesterol oxidation and decomposition, can disrupt redox homeostasis, impose stress and subsequently lead to accumulation of unfolded or misfolded proteins in brain cells. Alzheimer's (AD), Parkinson's (PD), Huntington's disease (HD), Amyothrophic lateral sclerosis (ALS) and Friedreich ataxia (FRDA) are major neurological disorders associated with production of abnormal proteins and, as such, belong to the so called "protein conformational diseases". The Central Nervous System has evolved highly specific signaling pathways called the unfolded protein response to cope with the accumulation of unfolded or misfolded proteins. Recent discoveries of the mechanisms of cellular stress signaling have led to major new insights into the diverse processes that are regulated by cellular stress response. Thus, the pathogenic dysfunctional aggregation of proteins in non-native conformations is associated with metabolic derangements and excessive production of ROS. The brain response to detect and control metabolic or oxidative stress is accomplished by a complex network of "longevity assurance processes" integrated to the expression of genes termed vitagenes. Heat shock proteins are a highly conserved system responsible for the preservation and repair of correct protein conformation. Heme oxygenase-1, a inducible and redox-regulated enzyme, is currently considered as having an important role in cellular antioxidant defense. A neuroprotective effect, due to its heme degrading activity, and tissue-specific antioxidant effects due to its products CO and biliverdin, this latter being further reduced by biliverdin reductase in bilirubin is an emerging concept. There is a current interest in dietary compounds that can inhibit, retard or reverse the multi-stage pathophysiology of Alzheimer disease, with a chronic inflammatory response, brain injury and beta-amyloid associated pathology. Curcumin and ferulic acid, two powerful antioxidants, the first from the curry spice turmeric and the second a major constituent of fruit and vegetables, have emerged as strong inducers of the heat shock response. Food supplementation with curcumin and ferulic acid is considered a nutritional approach to reduce oxidative damage and amyloid pathology in Alzheimer disease. This review summarizes the complex regulation of cellular stress signaling and its relevance to human physiology and disease.}, }
@article {pmid17316906, year = {2008}, author = {Wang, J and Martin, E and Gonzales, V and Borchelt, DR and Lee, MK}, title = {Differential regulation of small heat shock proteins in transgenic mouse models of neurodegenerative diseases.}, journal = {Neurobiology of aging}, volume = {29}, number = {4}, pages = {586-597}, pmid = {17316906}, issn = {1558-1497}, support = {NS38065/NS/NINDS NIH HHS/United States ; NS38377/NS/NINDS NIH HHS/United States ; NS044278/NS/NINDS NIH HHS/United States ; P01 NS038065/NS/NINDS NIH HHS/United States ; R56 NS038065/NS/NINDS NIH HHS/United States ; P50 NS038377/NS/NINDS NIH HHS/United States ; R01 NS 044278/NS/NINDS NIH HHS/United States ; R01 NS038065/NS/NINDS NIH HHS/United States ; R01 NS044278/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; *Disease Models, Animal ; Female ; Heat-Shock Proteins, Small/*physiology ; Humans ; Male ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Mice, Transgenic ; Neurodegenerative Diseases/genetics/*metabolism ; Up-Regulation/*physiology ; }, abstract = {Previously, several studies have demonstrated changes in the levels of small heat shock proteins (sHSP) in the transgenic mouse models of familial amyotrophic lateral sclerosis (fALS) linked to mutations in Cu/Zn superoxide dismutase. Here, we compared the expression of sHSPs in transgenic mouse models of fALS, Parkinson's disease (PD), dentato-rubral pallido-luysian atrophy (DRPLA) and Huntington's disease (HD); where the expression of mutant cDNA genes was under the transcriptional regulation of the mouse prion protein promoter. These models express G37R mutant Cu/Zn superoxide dismutase (SOD1G37R; fALS), A53T mutant alpha-synuclein (alpha-SynA53T; PD), full-length mutant atrophin-1-65Q, and htt-N171-82Q (huntingtin N-terminal fragment; HD). We found that the levels and solubilities of two sHSPs, Hsp25 and alpha B-crystallin, were differentially regulated in these mice. Levels of both Hsp25 and alpha B-crystallin were markedly increased in subgroups of glias at the affected regions of symptomatic SODG37R and alpha-SynA53T transgenic mice; abnormal deposits or cells intensely positive for alpha B-crystallin were observed in SODG37R mice. By contrast, neither sHSP was induced in spinal cords of htt-N171-82Q or atrophin-1-65Q mice, which do not develop astrocytosis or major motor neuron abnormalities. Interestingly, the levels of insoluble alpha B-crystallin in spinal cords gradually increased as a function of age in nontransgenic animals. In vitro, alpha B-crystallin was capable of suppressing the aggregation of alpha-SynA53T, as previously described for a truncated mutant SOD1. The transgenes in these mice are expressed highly in astrocytes and thus our results suggest a role for small heat shock proteins in protecting activated glial cells such as astrocytes in neurodegenerative diseases.}, }
@article {pmid17403032, year = {2007}, author = {Yamashita, H and Kawamata, J and Okawa, K and Kanki, R and Nakamizo, T and Hatayama, T and Yamanaka, K and Takahashi, R and Shimohama, S}, title = {Heat-shock protein 105 interacts with and suppresses aggregation of mutant Cu/Zn superoxide dismutase: clues to a possible strategy for treating ALS.}, journal = {Journal of neurochemistry}, volume = {102}, number = {5}, pages = {1497-1505}, doi = {10.1111/j.1471-4159.2007.04534.x}, pmid = {17403032}, issn = {0022-3042}, mesh = {Age Factors ; Analysis of Variance ; Animals ; Cell Line ; Gene Expression Regulation/*physiology ; HSP110 Heat-Shock Proteins/*metabolism ; Humans ; Immunoprecipitation ; Mass Spectrometry ; Mice ; Mice, Transgenic ; Motor Neurons/metabolism ; Mutation/*physiology ; Neuroblastoma ; Spinal Cord/cytology/metabolism ; Superoxide Dismutase/genetics/*metabolism ; Transfection ; }, abstract = {A dominant mutation in the gene for copper-zinc superoxide dismutase (SOD1) is the most frequent cause of the inherited form of amyotrophic lateral sclerosis. Mutant SOD1 provokes progressive degeneration of motor neurons by an unidentified acquired toxicity. Exploiting both affinity purification and mass spectrometry, we identified a novel interaction between heat-shock protein 105 (Hsp105) and mutant SOD1. We detected this interaction both in spinal cord extracts of mutant SOD1(G93A) transgenic mice and in cultured neuroblastoma cells. Expression of Hsp105, which is found in mouse motor neurons, was depressed in the spinal cords of SOD1(G93A) mice as disease progressed, while levels of expression of two other heat-shock proteins, Hsp70 and Hsp27, were elevated. Moreover, Hsp105 suppressed the formation of mutant SOD1-containing aggregates in cultured cells. These results suggest that techniques that raise levels of Hsp105 might be promising tools for alleviation of the mutant SOD1 toxicity.}, }
@article {pmid17441507, year = {2007}, author = {Chen, S and Brown, IR}, title = {Neuronal expression of constitutive heat shock proteins: implications for neurodegenerative diseases.}, journal = {Cell stress &amp; chaperones}, volume = {12}, number = {1}, pages = {51-58}, pmid = {17441507}, issn = {1355-8145}, mesh = {Animals ; *Gene Expression Regulation ; HSC70 Heat-Shock Proteins/*metabolism ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/*metabolism ; Hyperthermia, Induced ; Male ; Neoplasm Proteins/*metabolism ; Neurodegenerative Diseases/epidemiology/*metabolism ; Neurons/classification/cytology/*metabolism ; Rats ; Rats, Wistar ; }, abstract = {Neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis have been termed "protein misfolding disorders." These diseases differ widely in frequency and impact different classes of neurons. Heat shock proteins provide a line of defense against misfolded, aggregation-prone proteins and are among the most potent suppressors of neurodegeneration in animal models. Analysis of constitutively expressed heat shock proteins revealed variable levels of Hsc70 and Hsp27 in different classes of neurons in the adult rat brain. The differing levels of these constitutively expressed heat shock proteins in neuronal cell populations correlated with the relative frequencies of the previously mentioned neurodegenerative diseases.}, }
@article {pmid17494697, year = {2007}, author = {Adachi, H and Waza, M and Tokui, K and Katsuno, M and Minamiyama, M and Tanaka, F and Doyu, M and Sobue, G}, title = {CHIP overexpression reduces mutant androgen receptor protein and ameliorates phenotypes of the spinal and bulbar muscular atrophy transgenic mouse model.}, journal = {The Journal of neuroscience : the official journal of the Society for Neuroscience}, volume = {27}, number = {19}, pages = {5115-5126}, pmid = {17494697}, issn = {1529-2401}, mesh = {Animals ; Central Nervous System/*metabolism/physiopathology ; Disease Models, Animal ; Down-Regulation/genetics ; Female ; Gene Expression Regulation/physiology ; Genetic Predisposition to Disease/genetics ; Genetic Therapy/*methods ; Heat-Shock Proteins/metabolism ; Humans ; Inclusion Bodies/genetics/metabolism ; Intranuclear Inclusion Bodies/genetics/metabolism ; Male ; Mice ; Mice, Transgenic ; Motor Neurons/metabolism/pathology ; Muscular Atrophy, Spinal/*genetics/*metabolism/therapy ; Mutation/genetics ; Nerve Degeneration/genetics/metabolism/therapy ; Phenotype ; Proteasome Endopeptidase Complex/metabolism ; Protein Folding ; Receptors, Androgen/genetics/*metabolism ; Ubiquitin-Protein Ligases/*genetics ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is an inherited motor neuron disease caused by the expansion of a polyglutamine tract within the androgen receptor (AR). The pathologic features of SBMA are motor neuron loss in the spinal cord and brainstem and diffuse nuclear accumulation and nuclear inclusions of the mutant AR in the residual motor neurons and certain visceral organs. Many components of the ubiquitin-proteasome and molecular chaperones are also sequestered in the inclusions, suggesting that they may be actively engaged in an attempt to degrade or refold the mutant AR. C terminus of Hsc70 (heat shock cognate protein 70)-interacting protein (CHIP), a U-box type E3 ubiquitin ligase, has been shown to interact with heat shock protein 90 (Hsp90) or Hsp70 and ubiquitylates unfolded proteins trapped by molecular chaperones and degrades them. Here, we demonstrate that transient overexpression of CHIP in a neuronal cell model reduces the monomeric mutant AR more effectively than it does the wild type, suggesting that the mutant AR is more sensitive to CHIP than is the wild type. High expression of CHIP in an SBMA transgenic mouse model also ameliorated motor symptoms and inhibited neuronal nuclear accumulation of the mutant AR. When CHIP was overexpressed in transgenic SBMA mice, mutant AR was also preferentially degraded over wild-type AR. These findings suggest that CHIP overexpression ameliorates SBMA phenotypes in mice by reducing nuclear-localized mutant AR via enhanced mutant AR degradation. Thus, CHIP overexpression would provide a potential therapeutic avenue for SBMA.}, }
@article {pmid17583678, year = {2007}, author = {Fukada, Y and Yasui, K and Kitayama, M and Doi, K and Nakano, T and Watanabe, Y and Nakashima, K}, title = {Gene expression analysis of the murine model of amyotrophic lateral sclerosis: studies of the Leu126delTT mutation in SOD1.}, journal = {Brain research}, volume = {1160}, number = {}, pages = {1-10}, doi = {10.1016/j.brainres.2007.05.044}, pmid = {17583678}, issn = {0006-8993}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/pathology/*physiopathology ; Animals ; Cathepsin H ; Cathepsins/genetics/metabolism ; Crystallins/genetics/metabolism ; Cysteine Endopeptidases/genetics/metabolism ; Disease Models, Animal ; *Gene Deletion ; Gene Expression/*physiology ; Gene Expression Regulation/genetics ; Heat-Shock Proteins/genetics ; Leucine/*genetics ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Molecular Chaperones ; Neoplasm Proteins/genetics ; Oligonucleotide Array Sequence Analysis/methods ; Peptide Initiation Factors/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Spinal Cord/metabolism/pathology ; Superoxide Dismutase/*genetics ; Superoxide Dismutase-1 ; mu-Crystallins ; }, abstract = {The pathogenic events that lead to amyotrophic lateral sclerosis (ALS) have not been elucidated. We previously described familial amyotrophic lateral sclerosis (FALS) caused by a Leu126delTT mutation in the Cu/Zn superoxide dismutase gene (SOD1) and have produced transgenic mice (TgM) carrying the same mutation (SOD1(L126delTT) TgM), which exhibited distinct ALS-like motor symptoms and pathological findings. In this study, we analyzed gene expression in the spinal cord of SOD1(L126delTT) TgM by cDNA microarray. Eleven genes were upregulated and two genes downregulated in pre-symptomatic TgM. In post-symptomatic TgM, 54 genes were upregulated and four genes downregulated. We performed real-time polymerase chain reaction (PCR) analysis of 10 of the 54 upregulated genes in the post-symptomatic TgM. The results of real-time PCR were consistent with those obtained by microarray for micro-crystallin (Crym), heat shock protein 1 (Hspb1/HSP27), serine proteinase inhibitor clade A member 3N (Serpina3n), complement component 1q subcomponent beta polypeptide (C1qb), cathepsin H (Ctsh) and polyadenylate binding protein-interacting protein 1 (Paip1). In immunohistochemical analysis, Hsbp1/HSP27 and Ctsh expression levels were increased in reactive astrocytes at the ventral horn of the spinal cord in post-symptomatic TgM, as were Crym, some of Ctsh and Paip1 in microglial cells. Increased expression of those genes was not observed in the control mice. These four genes may be related to the pathogenesis of FALS, especially with regard to the progression of reactive astrocytes and the inflammatory response of microglial cells.}, }
@article {pmid17623484, year = {2007}, author = {Dierick, I and Irobi, J and Janssens, S and Theuns, J and Lemmens, R and Jacobs, A and Corsmit, E and Hersmus, N and Van Den Bosch, L and Robberecht, W and De Jonghe, P and Van Broeckhoven, C and Timmerman, V}, title = {Genetic variant in the HSPB1 promoter region impairs the HSP27 stress response.}, journal = {Human mutation}, volume = {28}, number = {8}, pages = {830}, doi = {10.1002/humu.9503}, pmid = {17623484}, issn = {1098-1004}, mesh = {Animals ; Base Sequence ; COS Cells ; Chlorocebus aethiops ; Consensus Sequence ; DNA Mutational Analysis ; Electrophoretic Mobility Shift Assay ; Female ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/*genetics ; Heat-Shock Response/*genetics ; Humans ; Male ; Middle Aged ; Molecular Chaperones ; Molecular Sequence Data ; Mutation/*genetics ; Neoplasm Proteins/*genetics ; Promoter Regions, Genetic ; Protein Binding ; Response Elements/genetics ; Transcription, Genetic ; }, abstract = {The 27 kDa heat shock protein 1 (HSP27) is a member of the ubiquitously expressed small heat shock protein family and has pleiotropic cytoprotective functions. Since HSP27 may act as a motor neuron survival factor, we analyzed the genetic contribution of the human HSPB1 gene (HSPB1) to the etiology of amyotrophic lateral sclerosis (ALS). In a cohort of sporadic ALS patients, we identified three rare genetic variations and one of which (c.-217T>C) targeted a conserved nucleotide of the Heat Shock Element (HSE) in the HSPB1 promoter. Since binding of Heat Shock Factor 1 (HSF1) to this HSE is essential for stress-induced transcription of HSPB1, we examined the effect of the c.-217C allele on transcriptional activity and HSF binding. The basal promoter activity of the HSPB1 c.-217C mutant allele decreased to 50% as compared to the wild-type promoter in neuronal and non-neuronal cells. Following heat shock, the HSE variant attenuated significantly the stress-related increase in transcription. Electrophoretic mobility shift assays demonstrated a dramatically reduced HSF-binding to the c.-217C mutant allele as compared to the c.-217T wild-type allele. In conclusion, our study underscores the importance of the c.-217T nucleotide for HSF binding and heat inducibility of HSPB1. Therefore, our study suggests that the functional HSPB1 variant may represent a genetic modifier in the pathogenesis of motor neuron disease; however, it is necessary to confirm this HSPB1 variant in additional ALS patients.}, }
@article {pmid17656567, year = {2007}, author = {Brown, IR}, title = {Heat shock proteins and protection of the nervous system.}, journal = {Annals of the New York Academy of Sciences}, volume = {1113}, number = {}, pages = {147-158}, doi = {10.1196/annals.1391.032}, pmid = {17656567}, issn = {0077-8923}, mesh = {Animals ; Heat-Shock Proteins/*physiology ; Humans ; Nervous System Diseases/metabolism/pathology/prevention &amp; control ; *Nervous System Physiological Phenomena ; }, abstract = {Manipulation of the cellular stress response offers strategies to protect brain cells from damage induced by ischemia and neurodegenerative diseases. Overexpression of Hsp70 reduced ischemic injury in the mammalian brain. Investigation of the domains within Hsp70 that confers ischemic neuroprotection revealed the importance of the carboxyl-terminal domain. Arimoclomol, a coinducer of heat shock proteins, delayed progression of amyotrophic lateral sclerosis (ALS) in a mouse model in which motor neurons in the spinal cord and motor cortex degenerate. Celastrol, a promising candidate as an agent to counter neurodegenerative diseases, induced expression of a set of Hsps in differentiated neurons grown in tissue culture. Heat shock "preconditioning" protected the nervous system at the functional level of the synapse and selective overexpression of Hsp70 enhanced the level of synaptic protection. Following hyperthermia, constitutively expressed Hsc70 increased in synapse-rich areas of the brain where it associates with Hsp40 to form a complex that can refold denatured proteins. Stress tolerance in neurons is not solely dependent on their own Hsps but can be supplemented by Hsps from adjacent glial cells. Hence, application of exogenous Hsps at neural injury sites is an effective strategy to maintain neuronal viability.}, }
@article {pmid17688194, year = {2007}, author = {Taylor, DM and Tradewell, ML and Minotti, S and Durham, HD}, title = {Characterizing the role of Hsp90 in production of heat shock proteins in motor neurons reveals a suppressive effect of wild-type Hsf1.}, journal = {Cell stress &amp; chaperones}, volume = {12}, number = {2}, pages = {151-162}, pmid = {17688194}, issn = {1355-8145}, mesh = {Animals ; Carrier Proteins/metabolism ; Co-Repressor Proteins ; DNA-Binding Proteins/*metabolism ; HSP40 Heat-Shock Proteins/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; HSP90 Heat-Shock Proteins/antagonists &amp; inhibitors/*metabolism ; Heat Shock Transcription Factors ; Humans ; Hyperthermia, Induced ; Intracellular Signaling Peptides and Proteins/metabolism ; Mice ; Molecular Chaperones ; Motor Neurons/cytology/*metabolism ; Nuclear Proteins/metabolism ; Promoter Regions, Genetic/genetics ; Transcription Factors/*metabolism ; Up-Regulation/genetics ; }, abstract = {Induction of heat shock proteins (Hsps) is under investigation as treatment for neurodegenerative disorders, yet many types of neurons, including motor neurons that degenerate in amyotrophic lateral sclerosis (ALS), have a high threshold for activation of the major transcription factor mediating stress-induced Hsp upregulation, heat shock transcription factor 1 (Hsf1). Hsf1 is tightly regulated by a series of inhibitory checkpoints that include sequestration in multichaperone complexes governed by Hsp90. This study examined the role of multichaperone complexes in governing the heat shock response in motor neurons. Hsp90 inhibitors induced expression of Hsp70 and Hsp40 and transactivation of a human inducible hsp70 promoter-green fluorescent protein (GFP) reporter construct in motor neurons of dissociated spinal cord-dorsal root ganglion (DRG) cultures. On the other hand, overexpression of activator of Hsp90 adenosine triphosphatase ([ATPase 1], Aha1), which should mobilize Hsf1 by accelerating turnover of mature, adenosine triphosphate-(ATP) bound Hsp90 complexes, and death domain-associated protein (Daxx), which in cell lines has been shown to promote transcription of heat shock genes by relieving inhibition exerted by interactions between nuclear Hsp90/multichaperone complexes and trimeric Hsf1, failed to induce Hsps in the absence or presence of heat shock. These results indicate that disruption of multichaperone complexes alone is not sufficient to activate the neuronal heat shock response. Furthermore, in motor neurons, induction of Hsp70 by Hsp90-inhibiting drugs was prevented by overexpression of wild-type Hsfl, contrary to what would be expected for a classical Hsf1-mediated pathway. These results point to additional differences in regulation of hsp genes in neuronal and nonneuronal cells.}, }
@article {pmid17915556, year = {2007}, author = {Chow, AM and Brown, IR}, title = {Induction of heat shock proteins in differentiated human and rodent neurons by celastrol.}, journal = {Cell stress &amp; chaperones}, volume = {12}, number = {3}, pages = {237-244}, pmid = {17915556}, issn = {1355-8145}, mesh = {Animals ; *Cell Differentiation ; Cell Line ; Cell Survival/drug effects ; Dose-Response Relationship, Drug ; HSP70 Heat-Shock Proteins/metabolism ; Heat-Shock Proteins/*metabolism ; Humans ; Neurons/*drug effects/metabolism ; Neuroprotective Agents/*pharmacology ; Pentacyclic Triterpenes ; Rodentia ; Species Specificity ; Triterpenes/*pharmacology ; Up-Regulation ; }, abstract = {Neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis have been termed protein misfolding disorders that are characterized by the neuronal accumulation of protein aggregates. Manipulation of the cellular stress-response involving induction of heat shock proteins (Hsps) in differentiated neurons offers a therapeutic strategy to counter conformational changes in neuronal proteins that trigger pathogenic cascades resulting in neurodegenerative diseases. Hsps are protein repair agents that provide a line of defense against misfolded, aggregation-prone proteins. These proteins are not induced in differentiated neurons by conventional heat shock. We have found that celastrol, a quinine methide triterpene, induced expression of a wider set of Hsps, including Hsp70B', in differentiated human neurons grown in tissue culture compared to cultured rodent neuronal cells. Hence the beneficial effect of celastrol against human neurodegenerative diseases may exceed its potential in rodent models of these diseases.}, }
@article {pmid18045911, year = {2007}, author = {Gifondorwa, DJ and Robinson, MB and Hayes, CD and Taylor, AR and Prevette, DM and Oppenheim, RW and Caress, J and Milligan, CE}, title = {Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis.}, journal = {The Journal of neuroscience : the official journal of the Society for Neuroscience}, volume = {27}, number = {48}, pages = {13173-13180}, pmid = {18045911}, issn = {1529-2401}, support = {NS36081/NS/NINDS NIH HHS/United States ; NS46615/NS/NINDS NIH HHS/United States ; R01 NS036081/NS/NINDS NIH HHS/United States ; R29 NS036081/NS/NINDS NIH HHS/United States ; R01 NS046615/NS/NINDS NIH HHS/United States ; }, mesh = {Age Factors ; Amyotrophic Lateral Sclerosis/*drug therapy/*mortality/pathology/physiopathology ; Analysis of Variance ; Animals ; Behavior, Animal ; *Disease Models, Animal ; HSP70 Heat-Shock Proteins/*administration &amp; dosage/metabolism ; Hindlimb/pathology ; Mice ; Mice, Transgenic ; Motor Activity/drug effects ; Motor Neurons/drug effects ; Neuromuscular Junction ; Neuroprotective Agents/*therapeutic use ; Riluzole/therapeutic use ; Spinal Cord/drug effects/pathology ; Superoxide Dismutase/genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative disorder that results in the progressive loss of motoneurons (MNs) in the CNS. Several survival and death mechanisms of MNs have been characterized and it has been determined that MNs do not appear to mount a complete stress response, as determined by the lack of heat shock protein 70 (Hsp70) upregulation after several stress paradigms. Hsp70 has been shown to confer neuroprotection and the insufficient availability of Hsp70 may contribute to MNs' susceptibility to death in ALS mice. In this study, recombinant human Hsp70 (rhHsp70) was intraperitoneally injected three times weekly, beginning at postnatal day 50 until endstage, to G93A mutant SOD1 (G93A SOD1) mice. The administration of rhHsp70 was effective at increasing lifespan, delaying symptom onset, preserving motor function and prolonging MN survival. Interestingly, injected rhHsp70 localized to skeletal muscle and was not readily detected in the CNS. Treatment with rhHsp70 also resulted in an increased number of innervated neuromuscular junctions compared with control tissue. Together these results suggest rhHsp70 may delay disease progression in the G93A SOD1 mouse via a yet to be identified peripheral mechanism.}, }
@article {pmid18255302, year = {2008}, author = {Sharp, PS and Akbar, MT and Bouri, S and Senda, A and Joshi, K and Chen, HJ and Latchman, DS and Wells, DJ and de Belleroche, J}, title = {Protective effects of heat shock protein 27 in a model of ALS occur in the early stages of disease progression.}, journal = {Neurobiology of disease}, volume = {30}, number = {1}, pages = {42-55}, doi = {10.1016/j.nbd.2007.12.002}, pmid = {18255302}, issn = {1095-953X}, mesh = {Age Factors ; Amyotrophic Lateral Sclerosis/*metabolism/pathology ; Analysis of Variance ; Animals ; Behavior, Animal ; Cell Death ; Choline O-Acetyltransferase/metabolism ; Disease Models, Animal ; Disease Progression ; Gene Expression Regulation/*genetics ; Heat-Shock Proteins/genetics/*metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Molecular Chaperones ; Motor Neurons/*physiology ; Neoplasm Proteins/genetics/metabolism ; Psychomotor Performance ; RNA, Messenger/metabolism ; Reaction Time/genetics ; Spinal Cord/pathology ; Superoxide Dismutase/genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disorder, characterised by progressive motor neuron degeneration and muscle paralysis. Heat shock proteins (HSPs) have significant cytoprotective properties in several models of neurodegeneration. To investigate the therapeutic potential of heat shock protein 27 (HSP27) in a mouse model of ALS, we conducted an extensive characterisation of transgenic mice generated from a cross between HSP27 overexpressing mice and mice expressing mutant superoxide dismutase (SOD1(G93A)). We report that SOD1(G93A)/HSP27 double transgenic mice showed delayed decline in motor strength, a significant improvement in the number of functional motor units and increased survival of spinal motor neurons compared to SOD1(G93A) single transgenics during the early phase of disease. However, there was no evidence of sustained neuroprotection affecting long-term survival. Marked down-regulation of HSP27 protein occurred during disease progression that was not associated with a reduction in HSP27 mRNA, indicating a translational dysfunction due to the presence of mutant SOD1 protein. This study provides further support for the therapeutic potential of HSPs in ALS and other motor neuron disorders.}, }
@article {pmid18319614, year = {2008}, author = {An, JJ and Lee, YP and Kim, SY and Lee, SH and Kim, DW and Lee, MJ and Jeong, MS and Jang, SH and Kang, JH and Kwon, HY and Kang, TC and Won, MH and Cho, SW and Kwon, OS and Lee, KS and Park, J and Eum, WS and Choi, SY}, title = {Transduction of familial amyotrophic lateral sclerosis-related mutant PEP-1-SOD proteins into neuronal cells.}, journal = {Molecules and cells}, volume = {25}, number = {1}, pages = {55-63}, pmid = {18319614}, issn = {1016-8478}, mesh = {*Amyotrophic Lateral Sclerosis/genetics/metabolism ; Animals ; Astrocytes/cytology/*physiology ; Cell Survival ; Cells, Cultured ; Cysteamine/*analogs &amp; derivatives/metabolism ; Heat-Shock Proteins/genetics/metabolism ; Humans ; Oxidative Stress ; Peptides/genetics/*metabolism ; Rats ; Rats, Wistar ; Recombinant Fusion Proteins/genetics/*metabolism ; Superoxide Dismutase/genetics/*metabolism ; Superoxide Dismutase-1 ; *Transduction, Genetic ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.}, }
@article {pmid18325928, year = {2008}, author = {Dierick, I and Baets, J and Irobi, J and Jacobs, A and De Vriendt, E and Deconinck, T and Merlini, L and Van den Bergh, P and Rasic, VM and Robberecht, W and Fischer, D and Morales, RJ and Mitrovic, Z and Seeman, P and Mazanec, R and Kochanski, A and Jordanova, A and Auer-Grumbach, M and Helderman-van den Enden, AT and Wokke, JH and Nelis, E and De Jonghe, P and Timmerman, V}, title = {Relative contribution of mutations in genes for autosomal dominant distal hereditary motor neuropathies: a genotype-phenotype correlation study.}, journal = {Brain : a journal of neurology}, volume = {131}, number = {Pt 5}, pages = {1217-1227}, doi = {10.1093/brain/awn029}, pmid = {18325928}, issn = {1460-2156}, mesh = {Base Sequence ; Chromosomes, Human, Pair 11/genetics ; DNA Helicases ; Electrophysiology ; Female ; GTP-Binding Protein gamma Subunits/genetics ; Genotype ; HSP27 Heat-Shock Proteins ; Haplotypes ; Heat-Shock Proteins/genetics ; Hereditary Sensory and Motor Neuropathy/*genetics/physiopathology ; Humans ; Male ; Molecular Chaperones ; Mosaicism ; Multifunctional Enzymes ; *Mutation, Missense ; Neoplasm Proteins/genetics ; Pedigree ; Phenotype ; Protein Serine-Threonine Kinases/genetics ; RNA Helicases/genetics ; }, abstract = {Distal hereditary motor neuropathy (HMN) is a clinically and genetically heterogeneous group of disorders affecting spinal alpha-motor neurons. Since 2001, mutations in six different genes have been identified for autosomal dominant distal HMN; glycyl-tRNA synthetase (GARS), dynactin 1 (DCTN1), small heat shock 27 kDa protein 1 (HSPB1), small heat shock 22 kDa protein 8 (HSPB8), Berardinelli-Seip congenital lipodystrophy (BSCL2) and senataxin (SETX). In addition a mutation in the (VAMP)-associated protein B and C (VAPB) was found in several Brazilian families with complex and atypical forms of autosomal dominantly inherited motor neuron disease. We have investigated the distribution of mutations in these seven genes in a cohort of 112 familial and isolated patients with a diagnosis of distal motor neuropathy and found nine different disease-causing mutations in HSPB8, HSPB1, BSCL2 and SETX in 17 patients of whom 10 have been previously reported. No mutations were found in GARS, DCTN1 and VAPB. The phenotypic features of patients with mutations in HSPB8, HSPB1, BSCL2 and SETX fit within the distal HMN classification, with only one exception; a C-terminal HSPB1-mutation was associated with upper motor neuron signs. Furthermore, we provide evidence for a genetic mosaicism in transmitting an HSPB1 mutation. This study, performed in a large cohort of familial and isolated distal HMN patients, clearly confirms the genetic and phenotypic heterogeneity of distal HMN and provides a basis for the development of algorithms for diagnostic mutation screening in this group of disorders.}, }
@article {pmid18551622, year = {2008}, author = {Cudkowicz, ME and Shefner, JM and Simpson, E and Grasso, D and Yu, H and Zhang, H and Shui, A and Schoenfeld, D and Brown, RH and Wieland, S and Barber, JR and , }, title = {Arimoclomol at dosages up to 300 mg/day is well tolerated and safe in amyotrophic lateral sclerosis.}, journal = {Muscle &amp; nerve}, volume = {38}, number = {1}, pages = {837-844}, doi = {10.1002/mus.21059}, pmid = {18551622}, issn = {0148-639X}, support = {5M01 RR 0082729/RR/NCRR NIH HHS/United States ; M01 RR 01066/RR/NCRR NIH HHS/United States ; M01 RR 01346/RR/NCRR NIH HHS/United States ; M01 RR 023940/RR/NCRR NIH HHS/United States ; M01 RR 10732/RR/NCRR NIH HHS/United States ; }, mesh = {Aged ; Amyotrophic Lateral Sclerosis/*drug therapy/metabolism ; Blood-Brain Barrier ; Dose-Response Relationship, Drug ; Double-Blind Method ; Female ; Humans ; Hydroxylamines/*administration &amp; dosage/*adverse effects/pharmacokinetics ; Male ; Middle Aged ; }, abstract = {Arimoclomol is an investigational drug for amyotrophic lateral sclerosis (ALS) that amplifies heat shock protein gene expression during cell stress. The objectives of the present study were to assess the safety, tolerability, and pharmacokinetics of arimoclomol in ALS. Eighty-four participants with ALS received arimoclomol at one of three oral doses (25, 50, or 100 mg three times daily) or placebo. The primary outcome measure was safety and tolerability. A subset of 44 participants provided serum and cerebrospinal fluid (CSF) samples for pharmacokinetic analysis. Participants who completed 12 weeks of treatment could enroll in a 6-month open-label study. Arimoclomol at doses up to 300 mg/day was well tolerated and safe. Arimoclomol resulted in dose-linear pharmacologic exposures and the half-life did not change with continued treatment. Arimoclomol CSF levels increased with dose. Arimoclomol was shown to be safe, and it crosses the blood-brain barrier. Serum pharmacokinetic profiles support dosing of three times per day. An efficacy study in ALS is planned.}, }
@article {pmid18624915, year = {2008}, author = {Krishnan, J and Vannuvel, K and Andries, M and Waelkens, E and Robberecht, W and Van Den Bosch, L}, title = {Over-expression of Hsp27 does not influence disease in the mutant SOD1(G93A) mouse model of amyotrophic lateral sclerosis.}, journal = {Journal of neurochemistry}, volume = {106}, number = {5}, pages = {2170-2183}, doi = {10.1111/j.1471-4159.2008.05545.x}, pmid = {18624915}, issn = {1471-4159}, mesh = {Age of Onset ; Amyotrophic Lateral Sclerosis/genetics/*metabolism/physiopathology ; Animals ; Cell Survival/genetics ; Crosses, Genetic ; Cytoprotection/*genetics ; Disease Models, Animal ; Gene Expression Regulation/physiology ; Genetic Predisposition to Disease/genetics ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/genetics/*metabolism ; Humans ; Longevity/genetics ; Mice ; Mice, Transgenic ; Molecular Chaperones/genetics/*metabolism ; Neoplasm Proteins/genetics/*metabolism ; Spinal Cord/*metabolism/physiopathology ; Spinal Cord Ischemia/genetics/metabolism/physiopathology ; Superoxide Dismutase/genetics/*metabolism ; Superoxide Dismutase-1 ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a chronic, adult-onset neurodegenerative disorder characterized by the selective loss of upper and lower motor neurons, resulting in severe atrophy of muscles and death. Although the exact pathogenic mechanism of mutant superoxide dismutase 1 (SOD1) causing familial ALS is still elusive, toxic protein aggregation leading to insufficiency of chaperones is one of the main hypotheses. In this study, we investigated the effect of over-expressing one of these chaperones, heat shock protein 27 (Hsp27), in ALS. Mice over-expressing the human, mutant SOD1(G93A) were crossed with mice that ubiquitously over-expressed human Hsp27. Even though the single transgenic hHsp27 mice showed protection against spinal cord ischemia, the double transgenic SOD1(G93A)/hHsp27 mice did not live longer, and did not show a significant delay in the onset of disease compared to their SOD1(G93A) littermates. There was no protective effect of hHsp27 over-expression on the motor neurons and on the mutant SOD1 aggregates in the double transgenic SOD1(G93A)/hHsp27 mice. In conclusion, despite the protective action against acute motor neuron injury, Hsp27 alone is not sufficient to protect against the chronic motor neuron injury due to the presence of mutant SOD1.}, }
@article {pmid18635547, year = {2008}, author = {Rudrabhatla, P and Zheng, YL and Amin, ND and Kesavapany, S and Albers, W and Pant, HC}, title = {Pin1-dependent prolyl isomerization modulates the stress-induced phosphorylation of high molecular weight neurofilament protein.}, journal = {The Journal of biological chemistry}, volume = {283}, number = {39}, pages = {26737-26747}, pmid = {18635547}, issn = {0021-9258}, support = {//Intramural NIH HHS/United States ; }, mesh = {Adaptor Proteins, Signal Transducing/antagonists &amp; inhibitors/genetics/*metabolism ; Alzheimer Disease/genetics/metabolism/therapy ; Amyotrophic Lateral Sclerosis/genetics/metabolism/therapy ; Animals ; Apoptosis/drug effects/genetics ; Cell Line ; Cerebral Cortex/embryology/*metabolism ; *Heat-Shock Response/drug effects/genetics ; Humans ; Hydrogen Peroxide/pharmacology ; NIMA-Interacting Peptidylprolyl Isomerase ; Neurofilament Proteins/antagonists &amp; inhibitors/genetics/*metabolism ; Neurons/*metabolism ; Oxidants/pharmacology ; *Oxidative Stress/drug effects/genetics ; Peptidylprolyl Isomerase/antagonists &amp; inhibitors/genetics/*metabolism ; Phosphorylation/drug effects ; Protein Kinases/genetics/metabolism ; Rats ; Rats, Wistar ; tau Proteins/genetics/metabolism ; }, abstract = {Aberrant phosphorylation of neuronal cytoskeletal proteins is a key pathological event in neurodegenerative disorders such as Alzheimer disease (AD) and amyotrophic lateral sclerosis, but the underlying mechanisms are still unclear. Previous studies have shown that Pin1, a peptidylprolyl cis/trans-isomerase, may be actively involved in the regulation of Tau hyperphosphorylation in AD. Here, we show that Pin1 modulates oxidative stress-induced NF-H phosphorylation. In an in vitro kinase assay, the addition of Pin1 substantially increased phosphorylation of NF-H KSP repeats by proline-directed kinases, Erk1/2, Cdk5/p35, and JNK3 in a concentration-dependent manner. In vivo, dominant-negative (DN) Pin1 and Pin1 small interfering RNA inhibited epidermal growth factor-induced NF-H phosphorylation. Because oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases, we studied the role of Pin1 in stressed cortical neurons and HEK293 cells. Both hydrogen peroxide (H(2)O(2)) and heat stresses induce phosphorylation of NF-H in transfected HEK293 cells and primary cortical cultures. Knockdown of Pin1 by transfected Pin1 short interference RNA and DN-Pin1 rescues the effect of stress-induced NF-H phosphorylation. The H(2)O(2) and heat shock induced perikaryal phospho-NF-H accumulations, and neuronal apoptosis was rescued by inhibition of Pin1 in cortical neurons. JNK3, a brain-specific JNK isoform, is activated under oxidative and heat stresses, and inhibition of Pin1 by Pin1 short interference RNA and DN-Pin1 inhibits this pathway. These results implicate Pin1 as a possible modulator of stress-induced NF-H phosphorylation as seen in neurodegenerative disorders like AD and amyotrophic lateral sclerosis. Thus, Pin1 may be a potential therapeutic target for these diseases.}, }
@article {pmid18673445, year = {2008}, author = {Kalmar, B and Novoselov, S and Gray, A and Cheetham, ME and Margulis, B and Greensmith, L}, title = {Late stage treatment with arimoclomol delays disease progression and prevents protein aggregation in the SOD1 mouse model of ALS.}, journal = {Journal of neurochemistry}, volume = {107}, number = {2}, pages = {339-350}, doi = {10.1111/j.1471-4159.2008.05595.x}, pmid = {18673445}, issn = {1471-4159}, support = {G0401350/MRC_/Medical Research Council/United Kingdom ; G0601943/MRC_/Medical Research Council/United Kingdom ; G0700412/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Age Factors ; Amyotrophic Lateral Sclerosis/*drug therapy/genetics/*metabolism/pathology ; Animals ; Disease Models, Animal ; Disease Progression ; Female ; HSP70 Heat-Shock Proteins/*metabolism ; Humans ; Hydroxylamines/*therapeutic use ; Male ; Mice ; Mice, Transgenic ; Motor Neurons/drug effects/physiology ; Muscle, Skeletal/drug effects/pathology/physiopathology ; Spinal Cord/pathology ; Statistics, Nonparametric ; Superoxide Dismutase/genetics ; Survival Analysis ; Ubiquitin/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by motoneuron degeneration, resulting in muscle paralysis and death, typically within 1-5 years of diagnosis. Although the pathogenesis of ALS remains unclear, there is evidence for the involvement of proteasome dysfunction and heat shock proteins in the disease. We have previously shown that treatment with a co-inducer of the heat shock response called arimoclomol is effective in the SOD(G93A) mouse model of ALS, delaying disease progression and extending the lifespan of SOD(G93A) mice (Kieran et al. 2004). However, this previous study only examined the effects arimoclomol when treatment was initiated in pre- or early symptomatic stages of the disease. Clearly, to be of benefit to the majority of ALS patients, any therapy must be effective after symptom onset. In order to establish whether post-symptomatic treatment with arimoclomol is effective, in this study we carried out a systematic assessment of different treatment regimes in SOD(G93A) mice. Treatment with arimoclomol from early (75 days) or late (90 days) symptomatic stages significantly improved muscle function. Treatment from 75 days also significantly increased the lifespan of SOD(G93A) mice, although treatment from 90 days has no significant effect on lifespan. The mechanism of action of arimoclomol involves potentiation of the heat shock response, and treatment with arimoclomol increased Hsp70 expression. Interestingly, this up-regulation in Hsp70 was accompanied by a decrease in the number of ubiquitin-positive aggregates in the spinal cord of treated SOD(G93A) mice, suggesting that arimoclomol directly effects protein aggregation and degradation.}, }
@article {pmid18682744, year = {2008}, author = {Yang, J and Bridges, K and Chen, KY and Liu, AY}, title = {Riluzole increases the amount of latent HSF1 for an amplified heat shock response and cytoprotection.}, journal = {PloS one}, volume = {3}, number = {8}, pages = {e2864}, pmid = {18682744}, issn = {1932-6203}, mesh = {Cell Survival/drug effects ; DNA-Binding Proteins/biosynthesis/*genetics ; Excitatory Amino Acid Agonists/pharmacology ; Genes, Reporter ; HeLa Cells ; Heat Shock Transcription Factors ; Hot Temperature ; Humans ; Luciferases/genetics ; Neurodegenerative Diseases/drug therapy/physiopathology ; Neurons/drug effects/physiology ; Riluzole/*pharmacology ; Spinal Cord/embryology/physiology ; Transcription Factors/biosynthesis/*genetics ; }, abstract = {BACKGROUND: Induction of the heat shock response (HSR) and increased expression of the heat shock proteins (HSPs) provide mechanisms to ensure proper protein folding, trafficking, and disposition. The importance of HSPs is underscored by the understanding that protein mis-folding and aggregation contribute centrally to the pathogenesis of neurodegenerative diseases.<br><br>We used a cell-based hsp70-luciferease reporter gene assay system to identify agents that modulate the HSR and show here that clinically relevant concentrations of the FDA-approved ALS drug riluzole significantly increased the heat shock induction of hsp70-luciferse reporter gene. Immuno-Western and -cytochemical analysis of HSF1 show that riluzole increased the amount of cytosolic HSF1 to afford a greater activation of HSF1 upon heat shock. The increased HSF1 contributed centrally to the cytoprotective activity of riluzole as hsf1 gene knockout negated the synergistic activity of riluzole and conditioning heat shock to confer cell survival under oxidative stress. Evidence of a post-transcriptional mechanism for the increase in HSF1 include: quantitation of mRNA(hsf1) by RT-PCR showed no effect of either heat shock or riluzole treatment; riluzole also increased the expression of HSF1 from a CMV-promoter; analysis of the turnover of HSF1 by pulse chase and immunoprecipitation show that riluzole slowed the decay of [(35)S]labeled-HSF1. The effect of riluzole on HSF1 was qualitatively different from that of MG132 and chloroquine, inhibitors of the proteasome and lysosome, respectively, and appeared to involve the chaperone-mediated autophagy pathway as RNAi-mediated knockdown of CMA negated its effect.<br><br>CONCLUSION/SIGNIFICANCE: We show that riluzole increased the amount of HSF1 to amplify the HSR for cytoprotection. Our study provides novel insight into the mechanism that regulates HSF1 turnover, and identifies the degradation of HSF1 as a target for therapeutics intervention.}, }
@article {pmid18701194, year = {2010}, author = {Anagnostou, G and Akbar, MT and Paul, P and Angelinetta, C and Steiner, TJ and de Belleroche, J}, title = {Vesicle associated membrane protein B (VAPB) is decreased in ALS spinal cord.}, journal = {Neurobiology of aging}, volume = {31}, number = {6}, pages = {969-985}, doi = {10.1016/j.neurobiolaging.2008.07.005}, pmid = {18701194}, issn = {1558-1497}, mesh = {Adolescent ; Adult ; Age Factors ; Amyotrophic Lateral Sclerosis/metabolism/*pathology ; Caspase 1/genetics/metabolism ; Child ; Child, Preschool ; Female ; Gene Expression Regulation/*physiology ; HSP27 Heat-Shock Proteins/genetics/metabolism ; Heat-Shock Proteins/genetics/metabolism ; Humans ; Male ; Middle Aged ; Molecular Chaperones ; Motor Neurons/metabolism ; Nerve Tissue Proteins/genetics/metabolism ; Postmortem Changes ; Protein Serine-Threonine Kinases/genetics/metabolism ; RNA, Messenger/metabolism ; Sex Factors ; Spinal Cord/*metabolism/pathology ; Statistics, Nonparametric ; Vesicular Transport Proteins/genetics/*metabolism ; Young Adult ; }, abstract = {The aim of this study was to quantify spinal cord expression of genes known to cause familial amyotrophic lateral sclerosis (FALS) or influence survival in a large cohort of sporadic cases of ALS (SALS), in order to determine their relevance to pathogenic mechanisms occurring in SALS. The expression of superoxide dismutase 1 (SOD1), vesicle associated membrane protein (VAPB), senataxin (SETX), dynactin (DCTN1), vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF1), the small heat shock proteins, HSPB1 and HSPB8, and three genes activated during disease progression, caspases-1 and -3 and glial fibrillary acidic protein (GFAP), were quantified. Robust changes in the expression of four genes were found, VAPB mRNA levels were decreased in the spinal cord of ALS patients compared to controls (p<0.006), whilst HSPB1, HSPB8 and caspase-1 showed significant increases (1.5-2.3-fold). Expression of VAPB mRNA and protein was predominantly localised to large motor neurones further supporting the relevance of this finding to disease progression occurring in SALS.}, }
@article {pmid18955116, year = {2008}, author = {Rohde, G and Kermer, P and Reed, JC and Bähr, M and Weishaupt, JH}, title = {Neuron-specific overexpression of the co-chaperone Bcl-2-associated athanogene-1 in superoxide dismutase 1(G93A)-transgenic mice.}, journal = {Neuroscience}, volume = {157}, number = {4}, pages = {844-849}, doi = {10.1016/j.neuroscience.2008.09.055}, pmid = {18955116}, issn = {0306-4522}, mesh = {Age Factors ; Amyotrophic Lateral Sclerosis/genetics/metabolism/mortality/pathology ; Animals ; DNA-Binding Proteins/genetics/*metabolism ; Disease Models, Animal ; Gene Expression Regulation/*drug effects ; Humans ; Mice ; Mice, Transgenic ; Motor Activity/genetics ; Motor Neurons/*metabolism ; Phosphopyruvate Hydratase/metabolism ; Proto-Oncogene Proteins c-bcl-2/genetics/metabolism ; Spinal Cord/pathology ; Superoxide Dismutase/*genetics ; Survival Analysis ; Transcription Factors/genetics/*metabolism ; }, abstract = {Bcl-2-associated athanogene-1 (BAG1) binds heat-shock protein 70 (Hsp70)/Hsc70, increases intracellular chaperone activity in neurons and proved to be protective in several models for neurodegeneration. Mutations in the superoxide dismutase 1 (SOD1) gene account for approximately 20% of familial amyotrophic lateral sclerosis (ALS) cases. A common property shared by all mutant SOD1 (mtSOD1) species is abnormal protein folding and the propensity to form aggregates. Toxicity and aggregate formation of mutant SOD1 can be overcome by enhanced chaperone function in vitro. Moreover, expression of mtSOD1 decreases BAG1 levels in a motoneuronal cell line. Thus, several lines of evidence suggested a protective role of BAG1 in mtSOD1-mediated motoneuron degeneration. To explore the therapeutic potential of BAG1 in a model for ALS, we generated SOD1G93A/BAG1 double transgenic mice expressing BAG1 in a neuron-specific pattern. Surprisingly, substantially increased BAG1 protein levels in spinal cord neurons did not significantly alter the phenotype of SOD1G93A-transgenic mice. Hence, expression of BAG1 is not sufficient to protect against mtSOD1-induced motor dysfunction in vivo. Our work shows that, in contrast to the in vitro situation, modulation of multiple cellular functions in addition to enhanced expression of a single chaperone is required to protect against SOD1 toxicity, highlighting the necessity of combined treatment strategies for ALS.}, }
@article {pmid19183864, year = {2009}, author = {Kalmar, B and Greensmith, L}, title = {Activation of the heat shock response in a primary cellular model of motoneuron neurodegeneration-evidence for neuroprotective and neurotoxic effects.}, journal = {Cellular &amp; molecular biology letters}, volume = {14}, number = {2}, pages = {319-335}, pmid = {19183864}, issn = {1689-1392}, support = {G0601943/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Apoptosis ; Cells, Cultured ; HSP70 Heat-Shock Proteins/metabolism ; *Heat-Shock Response/drug effects ; Hydroxylamines/pharmacology ; Mice ; Motor Neurons/cytology/*metabolism ; Neuroprotective Agents/pharmacology ; Oxidative Stress ; Pentacyclic Triterpenes ; Rats ; Staurosporine/pharmacology ; Triterpenes/pharmacology ; }, abstract = {Pharmacological up-regulation of heat shock proteins (hsps) rescues motoneurons from cell death in a mouse model of amyotrophic lateral sclerosis. However, the relationship between increased hsp expression and neuronal survival is not straightforward. Here we examined the effects of two pharmacological agents that induce the heat shock response via activation of HSF-1, on stressed primary motoneurons in culture. Although both arimoclomol and celastrol induced the expression of Hsp70, their effects on primary motoneurons in culture were significantly different. Whereas arimoclomol had survival-promoting effects, rescuing motoneurons from staurosporin and H(2)O(2) induced apoptosis, celastrol not only failed to protect stressed motoneurons from apoptosis under same experimental conditions, but was neurotoxic and induced neuronal death. Immunostaining of celastrol-treated cultures for hsp70 and activated caspase-3 revealed that celastrol treatment activates both the heat shock response and the apoptotic cell death cascade. These results indicate that not all agents that activate the heat shock response will necessarily be neuroprotective.}, }
@article {pmid19335999, year = {2009}, author = {An, JJ and Lee, YP and Kim, DW and Sohn, EJ and Jeong, HJ and Kang, HW and Shin, MJ and Kim, MJ and Ahn, EH and Jang, SH and Kang, JH and Kang, TC and Won, MH and Kwon, OS and Cho, SW and Lee, KS and Park, J and Eum, WS and Choi, SY}, title = {Transduced HSP27 protein protects neuronal cell death by enhancing FALS-associated SOD1 mutant activity.}, journal = {BMB reports}, volume = {42}, number = {3}, pages = {136-141}, doi = {10.5483/bmbrep.2009.42.3.136}, pmid = {19335999}, issn = {1976-6696}, mesh = {Amyotrophic Lateral Sclerosis/*enzymology ; Astrocytes/cytology/metabolism ; Cell Death ; Cell Survival ; Cysteamine/analogs &amp; derivatives/metabolism ; *Cytoprotection ; HSP27 Heat-Shock Proteins/*metabolism ; Heat-Shock Proteins ; Humans ; Molecular Chaperones ; Mutant Proteins/*metabolism ; Neurons/*cytology/metabolism ; Oxidative Stress ; Peptides/metabolism ; Protein Structure, Quaternary ; Recombinant Fusion Proteins/isolation &amp; purification ; Superoxide Dismutase/chemistry/*metabolism ; Superoxide Dismutase-1 ; *Transduction, Genetic ; }, abstract = {Familial Amyotrophic lateral sclerosis (FALS) is a progressive neurodegenetative disorder induced by mutations of the SOD1 gene. Heat shock protein 27 (HSP27) is well-defined as a stress-inducible protein, however the its role in ALS protection has not yet been established. To investigate the role HSP27 may have in SOD1 mutant-mediated apoptosis, human SOD1 or HSP27 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame fusion protein, which was then transduced into cells. We found the purified PEP-1-HSP27 fusion proteins can be transduced efficiently into neuronal cells and protect against cell death by enhancing mutant SOD1 activity. Moreover, transduced PEP-1-HSP27 efficiently prevents protein aggregation produced by oxidative stress. These results suggest that transduced HSP27 fusion protein may be explored as a potential therapeutic agent for FALS patients.}, }
@article {pmid19399234, year = {2009}, author = {Banno, H and Katsuno, M and Suzuki, K and Tanaka, F and Sobue, G}, title = {Neuropathology and therapeutic intervention in spinal and bulbar muscular atrophy.}, journal = {International journal of molecular sciences}, volume = {10}, number = {3}, pages = {1000-1012}, pmid = {19399234}, issn = {1422-0067}, mesh = {Animals ; Antineoplastic Agents, Hormonal/therapeutic use ; Heat-Shock Proteins/metabolism ; Humans ; Leuprolide/therapeutic use ; Muscular Atrophy, Spinal/drug therapy/metabolism/*pathology ; Peptides/genetics/metabolism ; Receptors, Androgen/metabolism ; Testosterone/blood ; Trinucleotide Repeats ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is a hereditary motor neuron disease caused by the expansion of a polyglutamine tract in the androgen receptor (AR). The histopathological finding in SBMA is loss of lower motor neurons in the anterior horn of the spinal cord as well as in the brainstem motor nuclei. Animal studies have revealed that the pathogenesis of SBMA depends on the level of serum testosterone, and that androgen deprivation mitigates neurodegeneration through inhibition of nuclear accumulation of the pathogenic AR. Heat shock proteins, ubiquitin-proteasome system and transcriptional regulation are also potential targets of therapy development for SBMA.}, }
@article {pmid19486304, year = {2009}, author = {Suzuki, K and Kastuno, M and Banno, H and Sobue, G}, title = {Pathogenesis-targeting therapeutics for spinal and bulbar muscular atrophy (SBMA).}, journal = {Neuropathology : official journal of the Japanese Society of Neuropathology}, volume = {29}, number = {4}, pages = {509-516}, doi = {10.1111/j.1440-1789.2009.01013.x}, pmid = {19486304}, issn = {1440-1789}, mesh = {Animals ; Clinical Trials as Topic/methods ; Drug Delivery Systems/methods/*trends ; Gene Targeting/methods/*trends ; Humans ; Muscular Atrophy/*pathology/therapy ; Muscular Atrophy, Spinal/*pathology/therapy ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA) is an hereditary, adult-onset, lower motor neuron disease caused by an aberrant elongation of a trinucleotide CAG repeat, which encodes the polyglutamine tract, in the first exon of the androgen receptor (AR) gene. The main symptoms are slowly progressive muscle weakness and atrophy of bulbar, facial and limb muscles. The cardinal histopathological findings of SBMA are an extensive loss of lower motor neurons in the anterior horn of the spinal cord as well as in brainstem motor nuclei and intranuclear accumulations of mutant AR protein in the residual motor neurons. Androgen deprivation therapy rescues neuronal dysfunction in animal models of SBMA, suggesting that the molecular basis for motor neuron degeneration in this disorder is testosterone-dependent nuclear accumulation of the mutant AR. Suppression of disease progression by leuprorelin acetate has also been demonstrated in a phase 2 clinical trial. In addition, the clarification of pathophysiology leads to appearance of candidate drugs to treat this devastating disease: heat shock protein (HSP) inducer, Hsp90 inhibitor, and histone deacetylase inhibitor. Advances in basic and clinical research on SBMA are now paving the way for clinical application of pathogenesis-targeting therapeutics.}, }
@article {pmid19697878, year = {2009}, author = {Banno, H and Katsuno, M and Suzuki, K and Iguchi, Y and Adachi, H and Tanaka, F and Sobue, G}, title = {[Molecular-targeted therapy for motor neuron disease].}, journal = {Brain and nerve = Shinkei kenkyu no shinpo}, volume = {61}, number = {8}, pages = {891-900}, pmid = {19697878}, issn = {1881-6096}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/*therapy ; Animals ; Anti-Inflammatory Agents, Non-Steroidal/therapeutic use ; Butyrates/administration &amp; dosage ; DNA-Binding Proteins/genetics ; Dyslipidemias ; Free Radical Scavengers/therapeutic use ; Heat-Shock Proteins/antagonists &amp; inhibitors ; Histone Deacetylase Inhibitors ; Humans ; Leuprolide/therapeutic use ; Male ; Mice ; Muscular Disorders, Atrophic/*genetics/*therapy ; Mutation ; Nerve Growth Factors/therapeutic use ; Receptors, Androgen/physiology ; Superoxide Dismutase/genetics/physiology ; Superoxide Dismutase-1 ; }, abstract = {The mechanisms underlying selective motor neuron degeneration in amyotrophic lateral sclerosis (ALS) remain unknown. There have been several important clinical trials on the treatment of ALS and treatment efficacy studies using mouse (SOD1) models of ALS. The latter revealed that diminished mutant SOD1 expression in the astrocytes delayed microglial activation and slowed disease progression. Dyslipidemia has been reported to have a protective effect in ALS patients. Current evidence has implicated a 43-kDa TAR DNA-binding protein (TDP-43) in the pathologenesis of ALS. Several mutations in TDP-43 were discovered in families with inherited motor neuron disease. Although phase III trials revealed that creatine monohydrate and IGF-1 was not beneficial for patients with ALS, favorable outcomes in SOD1 mice were reported with lithium, NADPH oxidase inhibitor, free-radical scavenger, and ammonium tetrathiomolybdate. Spinal and bulbar muscular atrophy (SBMA) is an adult-onset motor neuron disease affecting only males. Animal studies have revealed that the pathogenesis of SBMA depends on the serum testosterone level and that androgen deprivation mitigates neurodegeneration through inhibition of nuclear accumulation of the pathogenic androgen receptor (AR). Our studies have also identified several candidates for the treatment of SBMA. Selective inhibition of heat shock protein (HSP) facilitates the proteasomal degradation of pathogenic AR, leading to improvements in the signs and symptoms of SBMA mice. Oral administration of sodium butyrate--a histone deacetylase inhibitor--resulted in the improvement of neurological dysfunction in the SBMA mouse model, although its therapeutic dose range is narrow.}, }
@article {pmid19777341, year = {2010}, author = {Miltiadous, P and Stamatakis, A and Stylianopoulou, F}, title = {Neuroprotective effects of IGF-I following kainic acid-induced hippocampal degeneration in the rat.}, journal = {Cellular and molecular neurobiology}, volume = {30}, number = {3}, pages = {347-360}, pmid = {19777341}, issn = {1573-6830}, mesh = {Animals ; Biomarkers/analysis/metabolism ; Cell Death/drug effects/physiology ; Cytoprotection/drug effects/physiology ; Dentate Gyrus/drug effects/pathology/physiopathology ; Disease Models, Animal ; Epilepsy, Temporal Lobe/chemically induced/*complications ; Fluoresceins ; Glial Fibrillary Acidic Protein/metabolism ; Gliosis/chemically induced/metabolism/physiopathology ; HSP72 Heat-Shock Proteins/drug effects/metabolism ; Hippocampus/*drug effects/pathology/physiopathology ; Insulin-Like Growth Factor I/*pharmacology/therapeutic use ; Kainic Acid/antagonists &amp; inhibitors/toxicity ; Male ; Nerve Degeneration/*drug therapy/physiopathology/prevention &amp; control ; Neurons/*drug effects/metabolism/pathology ; Neuroprotective Agents/*pharmacology/therapeutic use ; Neurotoxins/antagonists &amp; inhibitors/toxicity ; Organic Chemicals ; Rats ; Rats, Wistar ; Staining and Labeling ; Stress, Physiological/drug effects/physiology ; }, abstract = {Insulin-like growth factor I (IGF-I) has been shown to act as a neuroprotectant both in in vitro studies and in in vivo animal models of ischemia, hypoxia, trauma in the brain or the spinal cord, multiple and amyotrophic lateral sclerosis, Alzheimer's and Parkinson's disease. In the present study, we investigated the neuroprotective potential of IGF-I in the "kainic acid-induced degeneration of the hippocampus" model of temporal lobe epilepsy. Increased cell death--as detected by FluoroJade B staining--and extensive cell loss--as determined by cresyl violet staining--were observed mainly in the CA3 and CA4 areas of the ipsilateral and contralateral hippocampus, 7 days following intrahippocampal administration of kainic acid. Kainic acid injection also resulted in intense astrogliosis--as assessed by the number of glial fibrillary acidic protein (GFAP) immunopositive cells--in both hemispheres, forming a clear astroglial scar ipsilaterally to the injection site. Heat-shock protein 70 (Hsp70) immunopositive cells were also observed in the ipsilateral dentate gyrus (DG) following kainic acid injection. When IGF-I was administered together with kainic acid, practically no signs of degeneration were detected in the contralateral hemisphere, while in the ipsilateral, there was a smaller degree of cell loss, reduced number of FluoroJade B-stained cells, decreased reactive gliosis and fewer Hsp70-positive cells. Our present results extend further the cases in which IGF-I is shown to exhibit neuroprotective properties in neurodegenerative processes in the CNS.}, }
@article {pmid19853641, year = {2010}, author = {Brettschneider, J and Lehmensiek, V and Mogel, H and Pfeifle, M and Dorst, J and Hendrich, C and Ludolph, AC and Tumani, H}, title = {Proteome analysis reveals candidate markers of disease progression in amyotrophic lateral sclerosis (ALS).}, journal = {Neuroscience letters}, volume = {468}, number = {1}, pages = {23-27}, doi = {10.1016/j.neulet.2009.10.053}, pmid = {19853641}, issn = {1872-7972}, mesh = {Aged ; Amyotrophic Lateral Sclerosis/cerebrospinal fluid/*diagnosis ; Biomarkers/cerebrospinal fluid ; Disease Progression ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Middle Aged ; Prospective Studies ; Proteome/*metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; }, abstract = {OBJECTIVES: In amyotrophic lateral sclerosis (ALS) the pathological determinants of disease progression remain poorly understood. We aimed to identify a characteristic CSF protein pattern that could provide new candidate biomarkers of disease progression in ALS.<br><br>METHODS: Using the two-dimensional difference in gel electrophoresis (2-D-DIGE), we compared CSF samples from patients with ALS that showed a rapid progression of disease (ALS-rp, n=9) over a follow-up time of 2 years and from patients with ALS that showed a slow progression of disease over follow-up (ALS-sl, n=9) over the same period. Protein spots that showed significant differences between patients and controls were selected for further analysis by MALDI-TOF mass spectrometry. For validation of identified spots ELISA and nephelometry were performed for two candidate proteins on a second cohort of patients (n=40).<br><br>RESULTS: We identified 6 different proteins and their isoforms which were all upregulated in ALS-rp as compared to ALS-sl (heat shock protein1, alpha-1 antitrypsin, fetuin-A precursor, transferrin, transthyretin (TTR), nebulin-related anchoring protein). For Fetuin-A and TTR, our findings could be confirmed by quantitative assay.<br><br>CONCLUSIONS: Fetuin-A and TTR are promising candidate markers for disease progression in ALS that warrant further evaluation on a larger cohort of patients.}, }
@article {pmid19860665, year = {2009}, author = {Stetler, RA and Gao, Y and Signore, AP and Cao, G and Chen, J}, title = {HSP27: mechanisms of cellular protection against neuronal injury.}, journal = {Current molecular medicine}, volume = {9}, number = {7}, pages = {863-872}, pmid = {19860665}, issn = {1875-5666}, support = {NS43802/NS/NINDS NIH HHS/United States ; NS45048/NS/NINDS NIH HHS/United States ; R01 NS056118/NS/NINDS NIH HHS/United States ; R01 NS043802/NS/NINDS NIH HHS/United States ; NS36736/NS/NINDS NIH HHS/United States ; NS44178/NS/NINDS NIH HHS/United States ; R01 NS036736/NS/NINDS NIH HHS/United States ; R01 NS045048/NS/NINDS NIH HHS/United States ; NS56118/NS/NINDS NIH HHS/United States ; R01 NS044178/NS/NINDS NIH HHS/United States ; }, mesh = {Cell Death/physiology ; Gene Expression Regulation ; HSP27 Heat-Shock Proteins/genetics/*metabolism ; Heat-Shock Response/physiology ; Humans ; Nervous System Diseases/metabolism/pathology/physiopathology ; Neurons/metabolism/*pathology ; Neuroprotective Agents/*metabolism ; Protein Folding ; Protein Processing, Post-Translational ; Signal Transduction/physiology ; }, abstract = {The heat shock protein (HSP) family has long been associated with a generalized cellular stress response, particularly in terms of recognizing and chaperoning misfolded proteins. While HSPs in general appear to be protective, HSP27 has recently emerged as a particularly potent neuroprotectant in a number of diverse neurological disorders, ranging from ALS to stroke. Although its robust protective effect on a number of insults has been recognized, the mechanisms and regulation of HSP27's protective actions are still undergoing intense investigation. On the basis of recent studies, HSP27 appears to have a dynamic and diverse range of function in cellular survival. This review provides a forum to compare and contrast recent literature exploring the protective mechanism and regulation of HSP27, focusing on neurological disorders in particular, as they represent a range from protein aggregate-associated diseases to acute stress.}, }
@article {pmid19938902, year = {2009}, author = {Lanka, V and Wieland, S and Barber, J and Cudkowicz, M}, title = {Arimoclomol: a potential therapy under development for ALS.}, journal = {Expert opinion on investigational drugs}, volume = {18}, number = {12}, pages = {1907-1918}, doi = {10.1517/13543780903357486}, pmid = {19938902}, issn = {1744-7658}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy ; Animals ; Clinical Trials as Topic ; Cytoprotection/drug effects ; Drug Approval ; Drug Evaluation, Preclinical ; Heat-Shock Proteins/biosynthesis/drug effects ; Humans ; Hydroxylamines/adverse effects/pharmacokinetics/*pharmacology/*therapeutic use ; }, abstract = {Arimoclomol, an amplifier of heat shock protein expression involved in cellular stress response, has emerged as a potential therapeutic candidate in amyotrophic lateral sclerosis (ALS) in recent years. Treatment with arimoclomol was reported to improve survival and muscle function in a mouse model of motor neuron disease. Several single- and multiple-dose safety studies have been completed in healthy control subjects. A 3-month Phase IIa study in people with ALS demonstrated safety at dosages up to 300 mg/day and another study is currently recruiting participants with familial ALS caused by mutations in the superoxide dismutase gene. We review the rationale for testing arimoclomol in sporadic and familial ALS in the context of available safety and pharmacokinetic data. Published and unpublished literature relative to the drug in the past two decades is discussed. The current review attempts to bring together our existing understanding of the actions of arimoclomol with the disease profile of ALS. The pharmacological profile of arimoclomol and the available preclinical data make it a promising therapeutic possibility in ALS.}, }
@article {pmid20021225, year = {2009}, author = {Adachi, H and Katsuno, M and Waza, M and Minamiyama, M and Tanaka, F and Sobue, G}, title = {Heat shock proteins in neurodegenerative diseases: pathogenic roles and therapeutic implications.}, journal = {International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group}, volume = {25}, number = {8}, pages = {647-654}, doi = {10.3109/02656730903315823}, pmid = {20021225}, issn = {1464-5157}, mesh = {Animals ; Diterpenes/pharmacology ; HSP90 Heat-Shock Proteins/metabolism ; Heat-Shock Proteins/genetics/*physiology/*therapeutic use ; Mice ; Neurodegenerative Diseases/*drug therapy/*etiology ; Proteasome Endopeptidase Complex/metabolism ; Protein Folding ; Ubiquitin/therapeutic use ; }, abstract = {Neurodegenerative diseases including amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, and polyglutamine (polyQ) diseases are thought to be caused by protein misfolding. Heat shock proteins (HSPs), which function mainly as molecular chaperones, play an important role in the folding and quality control of proteins. The histopathological hallmark of neurodegenerative diseases is accumulation and/or inclusions of the disease-causing proteins in residual neurons in targeted regions of the nervous system. The inclusions combine with many components of molecular chaperone pathways and ubiquitin-proteasome, raising the possibility that misfolding and altered degradation of mutant proteins may be involved in the pathogenesis of neurodegenerative diseases. Overexpression of HSPs has been reported to reduce the number and size of inclusions and accumulation of disease-causing proteins, and ameliorate the phenotypes in neuronal cell and mouse models. Hsp90 inhibitors also exert therapeutic effects through selective proteasome degradation of its client proteins. Elucidation of its pathophysiology using animal models has led to the development of disease-modifying drugs, i.e., Hsp90 inhibitor and HSP inducer, which inhibit the pathogenic process of neuronal degeneration. These findings may provide the basis for development of an HSP-related therapy for neurodegenerative diseases.}, }
@article {pmid20084477, year = {2010}, author = {Chow, AM and Mok, P and Xiao, D and Khalouei, S and Brown, IR}, title = {Heteromeric complexes of heat shock protein 70 (HSP70) family members, including Hsp70B', in differentiated human neuronal cells.}, journal = {Cell stress &amp; chaperones}, volume = {15}, number = {5}, pages = {545-553}, pmid = {20084477}, issn = {1466-1268}, mesh = {Blotting, Western ; Cell Line ; HSP40 Heat-Shock Proteins/genetics/metabolism ; HSP70 Heat-Shock Proteins/genetics/*metabolism ; Humans ; Immunoprecipitation ; Neurons/*metabolism ; Protein Binding ; }, abstract = {Human neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis have been termed "protein misfolding disorders." Upregulation of heat shock proteins that target misfolded aggregation-prone proteins has been proposed as a potential therapeutic strategy to counter neurodegenerative disorders. The heat shock protein 70 (HSP70) family is well characterized for its cytoprotective effects against cell death and has been implicated in neuroprotection by overexpression studies. HSP70 family members exhibit sequence and structural conservation. The significance of the multiplicity of HSP70 proteins is unknown. In this study, coimmunoprecipitation was employed to determine if association of HSP70 family members occurs, including Hsp70B' which is present in the human genome but not in mouse and rat. Heteromeric complexes of Hsp70B', Hsp70, and Hsc70 were detected in differentiated human SH-SY5Y neuronal cells. Hsp70B' also formed complexes with Hsp40 suggesting a common co-chaperone for HSP70 family members.}, }
@article {pmid20166961, year = {2010}, author = {Sajjad, MU and Samson, B and Wyttenbach, A}, title = {Heat shock proteins: therapeutic drug targets for chronic neurodegeneration?.}, journal = {Current pharmaceutical biotechnology}, volume = {11}, number = {2}, pages = {198-215}, doi = {10.2174/138920110790909641}, pmid = {20166961}, issn = {1873-4316}, support = {G120/881/MRC_/Medical Research Council/United Kingdom ; /BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Brain/drug effects/metabolism ; Chronic Disease ; Heat-Shock Proteins/*biosynthesis/genetics/physiology ; Humans ; Molecular Structure ; Neurodegenerative Diseases/*drug therapy/genetics/metabolism ; Neuroprotective Agents/adverse effects/chemistry/*pharmacology/therapeutic use ; Prions/metabolism ; Protein Folding ; }, abstract = {Intra- and extracellular protein misfolding and aggregation is likely to contribute to a number of age-related central nervous system diseases ("proteinopathies"). Therefore, molecular chaperones, such as heat shock proteins (HSPs), that regulate protein folding, misfolding and adaption to cellular stress are emerging as therapeutic targets. Here we review the current knowledge of HSP-modulating drugs and discuss the opportunities and difficulties of their therapeutic use to treat proteinopathies such as Alzheimer's- and Parkinson's disease, the polyglutamine- and prion disorders and Amyotrophic Lateral Sclerosis.}, }
@article {pmid20166962, year = {2010}, author = {Nagai, Y and Fujikake, N and Popiel, HA and Wada, K}, title = {Induction of molecular chaperones as a therapeutic strategy for the polyglutamine diseases.}, journal = {Current pharmaceutical biotechnology}, volume = {11}, number = {2}, pages = {188-197}, doi = {10.2174/138920110790909650}, pmid = {20166962}, issn = {1873-4316}, mesh = {Amyloid/genetics/metabolism ; Animals ; Brain/metabolism ; Heredodegenerative Disorders, Nervous System/genetics/metabolism/*therapy ; Humans ; Molecular Chaperones/*biosynthesis/genetics/physiology ; Peptides/genetics/*metabolism ; Protein Folding ; }, abstract = {Protein misfolding and aggregation in the brain have been implicated as a common molecular pathogenesis of various neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and the polyglutamine (polyQ) diseases. The polyQ diseases are a group of nine hereditary neurodegenerative diseases, including Huntington's disease (HD) and various types of spinocerebellar ataxia (SCA), which are caused by abnormal expansions of the polyQ stretch (> 35-40 repeats) in unrelated disease-causative proteins. The expanded polyQ stretch is thought to trigger misfolding of these proteins, leading to their aggregation and accumulation as inclusion bodies in affected neurons, eventually resulting in neurodegeneration. Misfolding and aggregation of the polyQ protein are the most ideal therapeutic targets since they are the most upstream events in the pathogenic cascade, and therefore, therapeutic approaches using molecular chaperones, which prevent protein misfolding and assist the refolding of misfolded proteins, are being extensively investigated. Indeed, a variety of molecular chaperones such as Hsp70 and Hsp40 have been demonstrated to exert therapeutic effects against various experimental models of the polyQ diseases. Furthermore, toward developing pharmacological therapies, small chemical activators of heat shock transcription factor 1 (HSF1) such as geldanamycin and its derivative 17-AAG, which induce multiple endogenous molecular chaperones, have been proven to be effective not only in polyQ disease models, but also in other neurodegenerative disease models. We hope that brain-permeable molecular chaperone inducers will be developed as drugs against a wide range of neurodegenerative diseases in the near future.}, }
@article {pmid20170473, year = {2010}, author = {Arawaka, S and Machiya, Y and Kato, T}, title = {Heat shock proteins as suppressors of accumulation of toxic prefibrillar intermediates and misfolded proteins in neurodegenerative diseases.}, journal = {Current pharmaceutical biotechnology}, volume = {11}, number = {2}, pages = {158-166}, doi = {10.2174/138920110790909713}, pmid = {20170473}, issn = {1873-4316}, mesh = {Amyloid/*antagonists &amp; inhibitors/metabolism ; Animals ; Heat-Shock Proteins/genetics/metabolism/*physiology ; Humans ; Neurodegenerative Diseases/*metabolism ; Protein Folding ; Protein Structure, Secondary ; Solubility ; }, abstract = {The most characteristic feature of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and Huntington's disease, is the occurrence of extra- or intracellular fibrillar aggregates containing misfolded proteins with beta-sheet conformation. These aggregates are composed of distinct proteins in each neurodegenerative disease. However, mutations in genes encoding major constituents of aggregates, such as Abeta, tau, alpha-synuclein, SOD1 and huntingtin, have been identified to causally associate with familial forms of the diseases. Biochemical studies demonstrate that these mutant and some wild-type proteins tend to be misfolded or form aggregates. It has been proposed that these diseases are caused by a common mechanism involving misfolded proteins that trigger a toxic cascade leading to neuronal degeneration. This hypothesis is the basis of the therapeutic potential of heat shock proteins (HSPs), which prevent protein misfolding and aggregation. Transgenic animal models of the diseases have demonstrated that induction or overexpression of HSPs can suppress neuronal dysfunction and degeneration. Do the results promise clinical success for HSP-based therapy in neurodegenerative diseases? Recent findings regarding the pathogenic species generated during fibril formation have highlighted some of the beneficial and problematic aspects of HSP-based therapy. In this review, we focus on the pathogenic role of prefibrillar intermediates, including soluble oligomers and protofibrils, on neurodegeneration, and the relationship between prefibrillar intermediates and the proteins targeted by HSPs. We discuss in vitro and in vivo experimental data showing that HSPs counteract disease progression by acting as suppressors of toxic prefibrillar intermediates and toxic misfolded proteins in neurodegenerative diseases.}, }
@article {pmid20406180, year = {2010}, author = {Siciliano, G and Carlesi, C and Pasquali, L and Piazza, S and Pietracupa, S and Fornai, F and Ruggieri, S and Murri, L}, title = {Clinical trials for neuroprotection in ALS.}, journal = {CNS &amp; neurological disorders drug targets}, volume = {9}, number = {3}, pages = {305-313}, doi = {10.2174/187152710791292648}, pmid = {20406180}, issn = {1996-3181}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/genetics/metabolism/*pathology ; Animals ; Clinical Trials as Topic/*trends ; Humans ; Neuropharmacology/methods/*trends ; Neuroprotective Agents/*pharmacology/therapeutic use ; Neurotoxins/antagonists &amp; inhibitors ; }, abstract = {Owing to uncertainty on the pathogenic mechanisms underlying motor neuron degeneration in amyotrophic lateral sclerosis (ALS) riluzole remains the only available therapy, with only marginal effects on disease survival. Here we review some of the recent advances in the search for disease-modifying drugs for ALS based on their putative neuroprotective effetcs. A number of more or less established agents have recently been investigated also in ALS for their potential role in neuroprotection and relying on antiglutamatergic, antioxidant or antiapoptotic strategies. Among them Talampanel, beta-lactam antibiotics, Coenzyme Q10, and minocycline have been investigated. Progress has also been made in exploiting growth factors for the treatment of ALS, partly due to advances in developing effective delivery systems to the central nervous system. A number of new therapies have also been identified, including a novel class of compounds, such as heat-shock protein co-inducers, which upregulate cell stress responses, and agents promoting autophagy and mitochondriogenesis, such as lithium and rapamycin. More recently, alterations of mRNA processing were described as a pathogenic mechanism in genetically defined forms of ALS, as those related to TDP-43 and FUS-TLS gene mutations. This knowledge is expected to improve our understanding of the pathogenetic mechanism in ALS and developing more effective therapies.}, }
@article {pmid20538880, year = {2010}, author = {Irobi, J and Almeida-Souza, L and Asselbergh, B and De Winter, V and Goethals, S and Dierick, I and Krishnan, J and Timmermans, JP and Robberecht, W and De Jonghe, P and Van Den Bosch, L and Janssens, S and Timmerman, V}, title = {Mutant HSPB8 causes motor neuron-specific neurite degeneration.}, journal = {Human molecular genetics}, volume = {19}, number = {16}, pages = {3254-3265}, pmid = {20538880}, issn = {1460-2083}, mesh = {Amino Acid Substitution ; Amyloid beta-Protein Precursor/metabolism ; Animals ; Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Cells, Cultured ; DNA Damage ; Green Fluorescent Proteins/genetics/metabolism ; HSP20 Heat-Shock Proteins/genetics/*metabolism ; Heat-Shock Proteins ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Molecular Chaperones ; Motor Neurons/*metabolism/pathology ; Muscle Proteins/genetics/*metabolism ; *Mutation ; Neurites/*metabolism/pathology ; Neuroglia/metabolism ; Rats ; Rats, Wistar ; Transfection ; }, abstract = {Missense mutations (K141N and K141E) in the alpha-crystallin domain of the small heat shock protein HSPB8 (HSP22) cause distal hereditary motor neuropathy (distal HMN) or Charcot-Marie-Tooth neuropathy type 2L (CMT2L). The mechanism through which mutant HSPB8 leads to a specific motor neuron disease phenotype is currently unknown. To address this question, we compared the effect of mutant HSPB8 in primary neuronal and glial cell cultures. In motor neurons, expression of both HSPB8 K141N and K141E mutations clearly resulted in neurite degeneration, as manifested by a reduction in number of neurites per cell, as well as in a reduction in average length of the neurites. Furthermore, expression of the K141E (and to a lesser extent, K141N) mutation also induced spheroids in the neurites. We did not detect any signs of apoptosis in motor neurons, showing that mutant HSPB8 resulted in neurite degeneration without inducing neuronal death. While overt in motor neurons, these phenotypes were only very mildly present in sensory neurons and completely absent in cortical neurons. Also glial cells did not show an altered phenotype upon expression of mutant HSPB8. These findings show that despite the ubiquitous presence of HSPB8, only motor neurons appear to be affected by the K141N and K141E mutations which explain the predominant motor neuron phenotype in distal HMN and CMT2L.}, }
@article {pmid20570967, year = {2010}, author = {Crippa, V and Sau, D and Rusmini, P and Boncoraglio, A and Onesto, E and Bolzoni, E and Galbiati, M and Fontana, E and Marino, M and Carra, S and Bendotti, C and De Biasi, S and Poletti, A}, title = {The small heat shock protein B8 (HspB8) promotes autophagic removal of misfolded proteins involved in amyotrophic lateral sclerosis (ALS).}, journal = {Human molecular genetics}, volume = {19}, number = {17}, pages = {3440-3456}, doi = {10.1093/hmg/ddq257}, pmid = {20570967}, issn = {1460-2083}, support = {GGP06063/TI_/Telethon/Italy ; GGP07063/TI_/Telethon/Italy ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism/*physiopathology ; Animals ; *Autophagy ; Cell Line ; DNA-Binding Proteins/genetics/metabolism ; Female ; HSP20 Heat-Shock Proteins/genetics/*metabolism ; Heat-Shock Proteins/genetics/*metabolism ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Molecular Chaperones ; Motor Neurons/metabolism ; Muscle Proteins/genetics/*metabolism ; Protein Folding ; Protein Serine-Threonine Kinases/genetics/*metabolism ; Superoxide Dismutase/chemistry/genetics/*metabolism ; Superoxide Dismutase-1 ; }, abstract = {Several neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), are characterized by the presence of misfolded proteins, thought to trigger neurotoxicity. Some familial forms of ALS (fALS), clinically indistinguishable from sporadic ALS (sALS), are linked to superoxide dismutase 1 (SOD1) gene mutations. It has been shown that the mutant SOD1 misfolds, forms insoluble aggregates and impairs the proteasome. Using transgenic G93A-SOD1 mice, we found that spinal cord motor neurons, accumulating mutant SOD1 also over-express the small heat shock protein HspB8. Using motor neuronal fALS models, we demonstrated that HspB8 decreases aggregation and increases mutant SOD1 solubility and clearance, without affecting wild-type SOD1 turnover. Notably, HspB8 acts on mutant SOD1 even when the proteasome activity is specifically blocked. The pharmacological blockage of autophagy resulted in a dramatic increase of mutant SOD1 aggregates. Immunoprecipitation studies, performed during autophagic flux blockage, demonstrated that mutant SOD1 interacts with the HspB8/Bag3/Hsc70/CHIP multiheteromeric complex, known to selectively activate autophagic removal of misfolded proteins. Thus, HspB8 increases mutant SOD1 clearance via autophagy. Autophagy activation was also observed in lumbar spinal cord of transgenic G93A-SOD1 mice since several autophago-lysosomal structures were present in affected surviving motor neurons. Finally, we extended our observation to a different ALS model and demonstrated that HspB8 exerts similar effects on a truncated version of TDP-43, another protein involved both in fALS and in sALS. Overall, these results indicate that the pharmacological modulation of HspB8 expression in motor neurons may have important implications to unravel the molecular mechanisms involved both in fALS and in sALS.}, }
@article {pmid20582873, year = {2010}, author = {Phukan, J}, title = {Arimoclomol, a coinducer of heat shock proteins for the potential treatment of amyotrophic lateral sclerosis.}, journal = {IDrugs : the investigational drugs journal}, volume = {13}, number = {7}, pages = {482-496}, pmid = {20582873}, issn = {2040-3410}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/physiopathology ; Animals ; Chaperonins/agonists ; Drugs, Investigational/adverse effects/pharmacokinetics/*pharmacology/*therapeutic use ; Heat-Shock Proteins/*metabolism ; Heat-Shock Response/drug effects ; Humans ; Hydroxylamines/adverse effects/pharmacokinetics/*pharmacology/*therapeutic use ; Nerve Degeneration/drug therapy/prevention &amp; control ; Neurons/drug effects ; Neuroprotective Agents/adverse effects/pharmacokinetics/pharmacology/therapeutic use ; Stress, Physiological ; Up-Regulation/*drug effects ; }, abstract = {Recent years have seen an explosion of research into increasingly prevalent neurodegenerative diseases. Arimoclomol (BRX-220), being developed by CytRx Corp, is an oral therapeutic candidate for the treatment of amyotrophic lateral sclerosis (ALS), the most common form of motor neuron disease. ALS is a fatal, incurable disorder, which can present as sporadic (90 to 95% of cases) or familial (5 to 10% of cases) forms. The etiology of sporadic ALS remains unknown and much of the understanding of ALS pathogenesis has been derived through study of its familial forms; in particular, through study of autosomal dominant mutations in the SOD1 (copper/zinc superoxide dismutase) gene, which cause approximately 20% of familial ALS cases. Under conditions of excessive stress, arimoclomol induces amplification of the cytoprotective heat shock response in order to protect motor neurons from death. Comprehensive in vivo and in vitro studies demonstrated its effect in the prevention of neuronal loss and promotion of motor neuron survival, even after symptom onset. Clinical trials have reported good tolerability and safety. This paper discusses the rationale for arimoclomol use in ALS, the preclinical and clinical evidence collected to date, the likelihood of its promising preclinical results translating to humans, and the relevance of this research for neurodegeneration as a whole.}, }
@article {pmid20699327, year = {2010}, author = {Bosco, DA and Lemay, N and Ko, HK and Zhou, H and Burke, C and Kwiatkowski, TJ and Sapp, P and McKenna-Yasek, D and Brown, RH and Hayward, LJ}, title = {Mutant FUS proteins that cause amyotrophic lateral sclerosis incorporate into stress granules.}, journal = {Human molecular genetics}, volume = {19}, number = {21}, pages = {4160-4175}, pmid = {20699327}, issn = {1460-2083}, support = {1RC2NS070342/NS/NINDS NIH HHS/United States ; R01NS050557/NS/NINDS NIH HHS/United States ; U01NS05225/NS/NINDS NIH HHS/United States ; RC1 NS068391/NS/NINDS NIH HHS/United States ; 1RC1NS068391/NS/NINDS NIH HHS/United States ; }, mesh = {Adult ; Amyotrophic Lateral Sclerosis/*genetics ; Animals ; Cell Line ; Cytoplasm/metabolism ; Female ; Green Fluorescent Proteins/genetics ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Oxidative Stress ; RNA-Binding Protein FUS/genetics/metabolism/*physiology ; Zebrafish ; }, abstract = {Mutations in the RNA-binding protein FUS (fused in sarcoma) are linked to amyotrophic lateral sclerosis (ALS), but the mechanism by which these mutants cause motor neuron degeneration is not known. We report a novel ALS truncation mutant (R495X) that leads to a relatively severe ALS clinical phenotype compared with FUS missense mutations. Expression of R495X FUS, which abrogates a putative nuclear localization signal at the C-terminus of FUS, in HEK-293 cells and in the zebrafish spinal cord caused a striking cytoplasmic accumulation of the protein to a greater extent than that observed for recessive (H517Q) and dominant (R521G) missense mutants. Furthermore, in response to oxidative stress or heat shock conditions in cultures and in vivo, the ALS-linked FUS mutants, but not wild-type FUS, assembled into perinuclear stress granules in proportion to their cytoplasmic expression levels. These findings demonstrate a potential link between FUS mutations and cellular pathways involved in stress responses that may be relevant to altered motor neuron homeostasis in ALS.}, }
@article {pmid20699640, year = {2010}, author = {Crippa, V and Carra, S and Rusmini, P and Sau, D and Bolzoni, E and Bendotti, C and De Biasi, S and Poletti, A}, title = {A role of small heat shock protein B8 (HspB8) in the autophagic removal of misfolded proteins responsible for neurodegenerative diseases.}, journal = {Autophagy}, volume = {6}, number = {7}, pages = {958-960}, doi = {10.4161/auto.6.7.13042}, pmid = {20699640}, issn = {1554-8635}, mesh = {Amyotrophic Lateral Sclerosis/genetics/metabolism/pathology ; Animals ; Autophagy/*physiology ; Heat-Shock Proteins/*metabolism ; Humans ; Inclusion Bodies/metabolism ; Molecular Chaperones ; Motor Neurons/metabolism/pathology ; Neurodegenerative Diseases/genetics/*metabolism/pathology ; Proteasome Endopeptidase Complex/metabolism ; Protein Conformation ; *Protein Folding ; Protein Serine-Threonine Kinases/*metabolism ; Superoxide Dismutase/chemistry/genetics/metabolism ; Superoxide Dismutase-1 ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of upper and lower motor neurons. As with other age-dependent neurodegenerative disorders, ALS is linked to the presence of misfolded proteins that may perturb several intracellular mechanisms and trigger neurotoxicity. Misfolded proteins aggregate intracellularly generating insoluble inclusions that are a key neuropathological hallmark of ALS. Proteins involved in the intracellular degradative systems, signaling pathways and the human TAR DNA-binding protein TDP-43 are major components of these inclusions. While their role and cytotoxicity are still largely debated, aggregates represent a powerful marker to follow protein misfolding in the neurodegenerative processes. Using in vitro and in vivo models of mutant SOD1 associated familial ALS (fALS), we and other groups demonstrated that protein misfolding perturbs one of the major intracellular degradative pathways, the ubiquitin proteasome system, giving rise to a vicious cycle that leads to the further deposit of insoluble proteins and finally to the formation of inclusions. The aberrant response to mutated SOD1 thus leads to the activation of the cascade of events ultimately responsible for cell death. Hence, our idea is that, by assisting protein folding, we might reduce protein aggregation, restore a fully functional proteasome activity and/or other cascades of events triggered by the mutant proteins responsible for motor neuron death in ALS. This could be obtained by stimulating mutant protein turnover, using an alternative degradative pathway that could clear mutant SOD1, namely autophagy.}, }
@article {pmid20863580, year = {2010}, author = {Ranganathan, S and Fischbeck, KH}, title = {Therapeutic approaches to spinal and bulbar muscular atrophy.}, journal = {Trends in pharmacological sciences}, volume = {31}, number = {11}, pages = {523-527}, pmid = {20863580}, issn = {1873-3735}, support = {ZIA NS003037/ImNIH/Intramural NIH HHS/United States ; ZIA NS003038/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Androgen Antagonists/*therapeutic use ; Animals ; Female ; Gene Expression Regulation ; HSP90 Heat-Shock Proteins/genetics/metabolism ; Insulin-Like Growth Factor Binding Protein 1/metabolism ; Male ; Mice ; Mice, Transgenic ; Molecular Chaperones/metabolism ; Motor Neuron Disease/*drug therapy/genetics/metabolism/physiopathology ; Muscular Atrophy, Spinal/*drug therapy/genetics/metabolism/physiopathology ; Mutation ; Receptors, Androgen/*genetics/*metabolism ; Trinucleotide Repeat Expansion ; }, abstract = {Spinal and bulbar muscular atrophy is a hereditary motor neuron disease caused by trinucleotide repeat expansion in the androgen receptor gene. The disease mechanism probably involves a toxic gain of function in the mutant protein, because other mutations that cause a loss of androgen receptor function result in a different phenotype and the mutant protein is toxic in mouse models. In these models, the toxicity is ligand-dependent and is associated with protein aggregation, as well as altered transcriptional regulation, axonal transport and mitochondrial function. Various therapeutic approaches have shown efficacy in mouse models, including androgen reduction, heat shock protein 90 (HSP90) inhibition and insulin-like growth factor (IGF)-1 overexpression. Clinical trials of androgen-reducing agents have had mixed results, with indications of efficacy but no proof of clinically meaningful benefit to date. These clinical studies have established outcome measures for future trials of other agents that have been beneficial in animal studies.}, }
@article {pmid20886841, year = {2010}, author = {Herskowitz, JH and Seyfried, NT and Duong, DM and Xia, Q and Rees, HD and Gearing, M and Peng, J and Lah, JJ and Levey, AI}, title = {Phosphoproteomic analysis reveals site-specific changes in GFAP and NDRG2 phosphorylation in frontotemporal lobar degeneration.}, journal = {Journal of proteome research}, volume = {9}, number = {12}, pages = {6368-6379}, pmid = {20886841}, issn = {1535-3907}, support = {F32 AG032848/AG/NIA NIH HHS/United States ; P50 AG025688/AG/NIA NIH HHS/United States ; P50AG025688/AG/NIA NIH HHS/United States ; P30 NS055077/NS/NINDS NIH HHS/United States ; F32AG032848-02/AG/NIA NIH HHS/United States ; P30NS055077/NS/NINDS NIH HHS/United States ; }, mesh = {Aged ; Amino Acid Sequence ; Binding Sites ; Brain/metabolism/pathology ; Chromatography, Affinity ; Chromatography, Liquid ; DNA-Binding Proteins/analysis/metabolism ; Female ; Frontotemporal Lobar Degeneration/*metabolism/pathology ; Glial Fibrillary Acidic Protein/*analysis/metabolism ; Humans ; Immunoblotting ; Male ; Middle Aged ; Molecular Sequence Data ; Phosphopeptides/analysis/metabolism ; Phosphoproteins/*analysis/metabolism ; Phosphorylation ; Postmortem Changes ; Proteomics/*methods ; Serine/metabolism ; Tandem Mass Spectrometry ; Tumor Suppressor Proteins/*analysis/metabolism ; }, abstract = {Frontotemporal lobar degeneration (FTLD) is a progressive neurodegenerative disease characterized by behavioral abnormalities, personality changes, language dysfunction, and can co-occur with the development of motor neuron disease. One major pathological form of FTLD is characterized by intracellular deposition of ubiquitinated and phosphorylated TAR DNA binding protein-43 (TDP-43), suggesting that dysregulation in phosphorylation events may contribute to disease progression. However, to date systematic analysis of the phosphoproteome in FTLD brains has not been reported. In this study, we employed immobilized metal affinity chromatography (IMAC) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify phosphopeptides from FTLD and age-matched control post-mortem human brain tissue. Using this approach, we identified 786 phosphopeptides in frontal cortex (control and FTLD), in which the population of phosphopeptides represented approximately 50% of the total peptides analyzed. Label-free quantification using spectral counts revealed six proteins with significant changes in the FTLD phosphoproteome. N-myc-Downstream regulated gene 2 (NDRG2) and glial fibrillary acidic protein (GFAP) had an increased number of phosphospectra in FTLD, whereas microtubule associated protein 1A (MAP1A), reticulon 4 (RTN4; also referred to as neurite outgrowth inhibitor (Nogo)), protein kinase C gamma (PRKCG), and heat shock protein 90 kDa alpha, class A member 1(HSP90AA1) had significantly fewer phosphospectra compared to control brain. To validate these differences, we examined NDRG2 phosphorylation in FTLD brain by immunoblot analyses, and using a phosphoserine-13 (pSer13) GFAP monoclonal antibody we show an increase in pSer13 GFAP levels by immunoblot concomitant with increased overall GFAP levels in FTLD cases. These data highlight the utility of combining proteomic and phosphoproteomic strategies to characterize post-mortem human brain tissue.}, }
@article {pmid21071151, year = {2011}, author = {Krause, M and Rodrigues-Krause, Jda C}, title = {Extracellular heat shock proteins (eHSP70) in exercise: Possible targets outside the immune system and their role for neurodegenerative disorders treatment.}, journal = {Medical hypotheses}, volume = {76}, number = {2}, pages = {286-290}, doi = {10.1016/j.mehy.2010.10.025}, pmid = {21071151}, issn = {1532-2777}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Anti-Inflammatory Agents/pharmacology ; Axons/pathology ; *Exercise ; Exocytosis ; Glucose/metabolism ; HSP70 Heat-Shock Proteins/*metabolism ; HSP72 Heat-Shock Proteins/metabolism ; Humans ; Immune System ; Models, Biological ; Motor Neurons/metabolism/pathology ; Neurodegenerative Diseases/*metabolism/*therapy ; Oxidation-Reduction ; }, abstract = {The intracellular heat shock protein 70kDa (iHSP70) is a universal marker of stress protein whose expression is induced by different cell stressors, such as heat, metabolite deprivation, redox imbalances and also during physical exercise. The activation of the iHSP70 is sine qua non for the promotion of tissue repair, since the expression of this chaperone confers cytoprotection and also exerts anti-inflammatory effects. On the other hand, exercise also induces the appearance of HSP70 in the extracellular medium (eHSP70) but, so far, the eHSP70 function has been mainly attributed to the activation of the immune system, seeming to perform an opposite function from the iHSP70. Since a moderate intensity exercise bout induces a general anti-inflammatory response even in the presence of an elevated eHSP70, this protein could carry out other functions rather than immune activation. Because exercise generates heat and metabolic challenges (especially on glucose metabolism) we suggests that the motoneurons, a very active (possibly one of the most stressed cells during exercise) and also very sensitive cells to heat and glucose metabolism imbalances, could be the major sites for the eHSP70 function. Due to the importance of the iHSP70 for repair and stress adaptation, this protein must be present in abundance on the site of stress and, because of its intrinsic inability response to stress [low heat shock factor 1 (HSF-1) activation] and the structure of the motoneurons (very long cells), the iHSP70, produced on the very far nucleus, is not appropriately transported through the axon to the axon terminal, were it is required. Then, during the exercise, the released eHSP70 can be internalized by the motoneurons and act as intracellular chaperons, protecting this cell against oxidative damage, protein denaturation and many others. Since a decreased iHSP70 expression capacity is associated with neurodegeneration diseases (such as Parkinson, polyglutamine, Amyotrophic lateral sclerosis, Alzheimer's, Huntington's and many others), the understanding of the physiological function of the extracellular HSP70 could be helpful on the treatment of neurodegenerative and other neuronal diseases. Besides that, it could explain some of the beneficial effects of the pharmacological HSP70 activators and also the beneficial effects of the exercise among neuronal cells during neurodegenerative-inducing diseases.}, }
@article {pmid21296666, year = {2011}, author = {Tradewell, ML and Cooper, LA and Minotti, S and Durham, HD}, title = {Calcium dysregulation, mitochondrial pathology and protein aggregation in a culture model of amyotrophic lateral sclerosis: mechanistic relationship and differential sensitivity to intervention.}, journal = {Neurobiology of disease}, volume = {42}, number = {3}, pages = {265-275}, doi = {10.1016/j.nbd.2011.01.016}, pmid = {21296666}, issn = {1095-953X}, support = {//Canadian Institutes of Health Research/Canada ; }, mesh = {Amyotrophic Lateral Sclerosis/*metabolism/pathology ; Animals ; Calcium/*metabolism ; Cells, Cultured ; Disease Models, Animal ; Membrane Potential, Mitochondrial/physiology ; Mice ; Microscopy, Confocal ; Mitochondria/*metabolism/pathology ; Motor Neurons/*metabolism/pathology ; Proteasome Endopeptidase Complex/metabolism ; Spinal Cord/metabolism/pathology ; Superoxide Dismutase/metabolism ; Superoxide Dismutase-1 ; }, abstract = {The combination of Ca(2+) influx during neurotransmission and low cytosolic Ca(2+) buffering contributes to the preferential vulnerability of motor neurons in amyotrophic lateral sclerosis (ALS). This study investigated the relationship among Ca(2+) accumulation in intracellular compartments, mitochondrial abnormalities, and protein aggregation in a model of familial ALS (fALS1). Human SOD1, wild type (SOD1(WT)) or with the ALS-causing mutation G93A (SOD1(G93A)), was expressed in motor neurons of dissociated murine spinal cord-dorsal root ganglia (DRG) cultures. Elevation of mitochondrial Ca(2+) ([Ca(2+)](m)), decreased mitochondrial membrane potential (Δψ) and rounding of mitochondria occurred early, followed by increased endoplasmic reticular Ca(2+) ([Ca(2+)](ER)), elevated cytosolic Ca(2+) ([Ca(2+)](c)), and subsequent appearance of SOD1(G93A) inclusions (a consequence of protein aggregation). [Ca(2+)](c) was elevated to a greater extent in neurons with inclusions than in those with diffusely distributed SOD1(G93A) and promoted aggregation of mutant protein, not vice versa: both [Ca(2+)](c) and the percentage of neurons with SOD1(G93A) inclusions were reduced by co-expressing the cytosolic Ca(2+)-buffering protein, calbindin D-28K; treatment with the heat shock protein inducer, geldanamycin, prevented inclusions but not the increase in [Ca(2+)](c), [Ca(2+)](m) or loss of Δψ, and inhibiting proteasome activity with epoxomicin, known to promote aggregation of disease-causing mutant proteins including SOD1(G93A), had no effect on Ca(2+) levels. Both expression of SOD1(G93A) and epoxomicin-induced inhibition of proteasome activity caused mitochondrial rounding, independent of Ca(2+) dysregulation and reduced Δψ. That geldanamycin prevented inclusions and mitochondrial rounding, but not Ca(2+) dysregulation or loss of Δψ indicates that chaperone-based therapies to prevent protein aggregation may require co-therapy to address these other underlying mechanisms of toxicity.}, }
@article {pmid21355045, year = {2011}, author = {Zhang, T and Mullane, PC and Periz, G and Wang, J}, title = {TDP-43 neurotoxicity and protein aggregation modulated by heat shock factor and insulin/IGF-1 signaling.}, journal = {Human molecular genetics}, volume = {20}, number = {10}, pages = {1952-1965}, pmid = {21355045}, issn = {1460-2083}, support = {R00 NS062089/NS/NINDS NIH HHS/United States ; NS062089/NS/NINDS NIH HHS/United States ; }, mesh = {Aging/metabolism ; Animals ; Animals, Genetically Modified ; Caenorhabditis elegans/genetics/metabolism/*physiology ; DNA-Binding Proteins/genetics/*metabolism ; Disease Models, Animal ; Gene Expression Regulation ; HEK293 Cells ; Heat Shock Transcription Factors ; Heat-Shock Response ; Humans ; Insulin/*metabolism ; Insulin-Like Growth Factor I/*metabolism ; Intracellular Space/metabolism ; Models, Biological ; Neurons/metabolism/pathology ; Protein Multimerization ; Protein Transport ; *Signal Transduction ; TDP-43 Proteinopathies/physiopathology ; Temperature ; Transcription Factors/genetics/*metabolism ; }, abstract = {TAR DNA-binding protein 43 (TDP-43) plays a key role in the neurodegenerative diseases including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The nature of the TDP-43-mediated neurotoxicity associated with these diseases is not yet understood. Here, we have established transgenic Caenorhabditis elegans models that express human TDP-43 variants in the nervous system, including the full-length wild-type (WT) and mutant proteins and a pathologic C-terminal fragment. The C. elegans models developed severe locomotor defects associated with the aggregation of TDP-43 in neurons. In comparison to parallel Cu/Zn superoxide dismutase worm models, transgenic full-length TDP-43, including the WT protein, was highly neurotoxic. In addition, TDP-43 demonstrated an unusually high tendency to aggregate, a property intrinsic to the WT protein. The C-terminal 25 kDa fragment of TDP-43 was unstable but remarkably aggregation-prone. Distinct disulfide-linked TDP-43 dimers and oligomers were detected. In C. elegans, the neurotoxicity and the protein aggregation of TDP-43 were regulated by environmental temperature and heat shock transcriptional factor 1, indicating that a deficiency in protein quality control is a risk factor for TDP-43 proteinopathy. Furthermore, the neurotoxicity and the protein aggregation of TDP-43 can be significantly attenuated by a deficiency in the insulin/insulin-like growth factor 1 (IGF-1) signaling in C. elegans and mammalian cells. These results suggest that protein misfolding underlies the aging-dependent neurodegeneration associated with TDP-43 and that the insulin/IGF-1 signaling may be a target for therapies.}, }
@article {pmid21403864, year = {2011}, author = {Turturici, G and Sconzo, G and Geraci, F}, title = {Hsp70 and its molecular role in nervous system diseases.}, journal = {Biochemistry research international}, volume = {2011}, number = {}, pages = {618127}, pmid = {21403864}, issn = {2090-2255}, abstract = {Heat shock proteins (HSPs) are induced in response to many injuries including stroke, neurodegenerative disease, epilepsy, and trauma. The overexpression of one HSP in particular, Hsp70, serves a protective role in several different models of nervous system injury, but has also been linked to a deleterious role in some diseases. Hsp70 functions as a chaperone and protects neurons from protein aggregation and toxicity (Parkinson disease, Alzheimer disease, polyglutamine diseases, and amyotrophic lateral sclerosis), protects cells from apoptosis (Parkinson disease), is a stress marker (temporal lobe epilepsy), protects cells from inflammation (cerebral ischemic injury), has an adjuvant role in antigen presentation and is involved in the immune response in autoimmune disease (multiple sclerosis). The worldwide incidence of neurodegenerative diseases is high. As neurodegenerative diseases disproportionately affect older individuals, disease-related morbidity has increased along with the general increase in longevity. An understanding of the underlying mechanisms that lead to neurodegeneration is key to identifying methods of prevention and treatment. Investigators have observed protective effects of HSPs induced by preconditioning, overexpression, or drugs in a variety of models of brain disease. Experimental data suggest that manipulation of the cellular stress response may offer strategies to protect the brain during progression of neurodegenerative disease.}, }
@article {pmid21757921, year = {2011}, author = {Romi, F and Helgeland, G and Gilhus, NE}, title = {Heat-shock proteins in clinical neurology.}, journal = {European neurology}, volume = {66}, number = {2}, pages = {65-69}, doi = {10.1159/000329373}, pmid = {21757921}, issn = {1421-9913}, mesh = {Heat-Shock Proteins/*metabolism ; Humans ; Nervous System Diseases/classification/*metabolism/pathology ; *Neurology ; }, abstract = {Heat-shock proteins (HSPs) are antigen-presenting protein-aggregation-preventing chaperones, induced by cellular stress in eukaryotic cells. In this review, we focus on recent HSP advances in neurological disorders. In myasthenia gravis, patients responding to immunosuppressive therapy have reduced serum HSP-71 antibodies. Generalized and ocular myasthenia gravis patients have elevated serum HSP-70 antibodies, indicating common pathogenic mechanisms. In Guillain-Barré syndrome, HSP-70 antibodies are elevated in serum and cerebrospinal fluid, and serum levels are higher than in myasthenia gravis and multiple sclerosis. In multiple sclerosis, serum HSP-27 antibodies are elevated during relapses providing disease activation marker, while α,β-crystallin expression in brain lesions indicates remission phase initiation. In acute stroke, serum HSP-27 antibodies are elevated irrespective of stroke type and duration. In epilepsy, HSP-27 is induced in patients' astrocytes and cerebral blood vessel walls, and α,β-crystallin is expressed in epileptic foci. In neurodegenerative disorders such as Alzheimer dementia and Parkinson's disease, HSPs are upregulated in brain tissue, and α,β-crystallin modulates superoxide dismutase-1 (SOD-1) tissue accumulation in familial amyotrophic lateral sclerosis. HSPs play an important role in antigen-presentation and tolerance development. Antibody-mediated interference with their function alters immune responses causing neuropathology. The role of HSPs in clinical neurology should be the subject of future investigation.}, }
@article {pmid22129991, year = {2011}, author = {Neef, DW and Jaeger, AM and Thiele, DJ}, title = {Heat shock transcription factor 1 as a therapeutic target in neurodegenerative diseases.}, journal = {Nature reviews. Drug discovery}, volume = {10}, number = {12}, pages = {930-944}, pmid = {22129991}, issn = {1474-1784}, support = {R01-GM059911/GM/NIGMS NIH HHS/United States ; GM076954/GM/NIGMS NIH HHS/United States ; F32 GM076954/GM/NIGMS NIH HHS/United States ; R01 NS065890/NS/NINDS NIH HHS/United States ; R01 GM059911/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; DNA-Binding Proteins/antagonists &amp; inhibitors/genetics/*metabolism ; *Drug Delivery Systems/methods ; Heat Shock Transcription Factors ; Heat-Shock Proteins/antagonists &amp; inhibitors/genetics/*metabolism ; Humans ; Neurodegenerative Diseases/*drug therapy/*metabolism/pathology ; Transcription Factors/antagonists &amp; inhibitors/genetics/*metabolism ; }, abstract = {Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis and prion-based neurodegeneration are associated with the accumulation of misfolded proteins, resulting in neuronal dysfunction and cell death. However, current treatments for these diseases predominantly address disease symptoms, rather than the underlying protein misfolding and cell death, and are not able to halt or reverse the degenerative process. Studies in cell culture, fruitfly, worm and mouse models of protein misfolding-based neurodegenerative diseases indicate that enhancing the protein-folding capacity of cells, via elevated expression of chaperone proteins, has therapeutic potential. Here, we review advances in strategies to harness the power of the natural cellular protein-folding machinery through pharmacological activation of heat shock transcription factor 1--the master activator of chaperone protein gene expression--to treat neurodegenerative diseases.}, }
@article {pmid22232551, year = {2012}, author = {Zhang, T and Hwang, HY and Hao, H and Talbot, C and Wang, J}, title = {Caenorhabditis elegans RNA-processing protein TDP-1 regulates protein homeostasis and life span.}, journal = {The Journal of biological chemistry}, volume = {287}, number = {11}, pages = {8371-8382}, pmid = {22232551}, issn = {1083-351X}, support = {K99 NS062089/NS/NINDS NIH HHS/United States ; NS07432/NS/NINDS NIH HHS/United States ; R01 NS074324/NS/NINDS NIH HHS/United States ; NS062089/NS/NINDS NIH HHS/United States ; R00 NS062089/NS/NINDS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics/metabolism ; Animals ; Animals, Genetically Modified ; Caenorhabditis elegans/genetics/*metabolism ; Caenorhabditis elegans Proteins/genetics/metabolism ; DNA-Binding Proteins/*biosynthesis/genetics ; Forkhead Transcription Factors ; Frontotemporal Lobar Degeneration/genetics/metabolism ; Genetic Complementation Test ; Homeostasis/*physiology ; Humans ; Longevity/*physiology ; Mutation ; Transcription Factors/genetics/metabolism ; }, abstract = {Transactive response DNA-binding protein (TARDBP/TDP-43), a heterogeneous nuclear ribonucleoprotein (hnRNP) with diverse activities, is a common denominator in several neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Orthologs of TDP-43 exist in animals ranging from mammals to invertebrates. Here, we systematically studied mutant Caenorhabditis elegans lacking the nematode TDP-43 ortholog, TDP-1. Heterologous expression of human TDP-43 rescued the defects in C. elegans lacking TDP-1, suggesting their functions are conserved. Although the tdp-1 mutants exhibited deficits in fertility, growth, and locomotion, loss of tdp-1 attenuated defects in several C. elegans models of proteotoxicity. Loss of tdp-1 suppressed defects in transgenic C. elegans expressing TDP-43 or CuZn superoxide dismutase, both of which are associated with proteotoxicity in neurodegenerative diseases. Loss of tdp-1 also reduced defects in mutant animals lacking the heat shock factor HSF-1. Transcriptional profiling demonstrated that the loss of TDP-1 altered expression of genes functioning in RNA processing and protein folding. Furthermore, the absence of tdp-1 extended the life span in C. elegans. The life span extension required a FOXO transcriptional factor DAF-16 but not HSF-1. These results suggest that the C. elegans TDP-1 has a role in the regulation of protein homeostasis and aging.}, }
@article {pmid22353756, year = {2012}, author = {Mimoto, T and Miyazaki, K and Morimoto, N and Kurata, T and Satoh, K and Ikeda, Y and Abe, K}, title = {Impaired antioxydative Keap1/Nrf2 system and the downstream stress protein responses in the motor neuron of ALS model mice.}, journal = {Brain research}, volume = {1446}, number = {}, pages = {109-118}, doi = {10.1016/j.brainres.2011.12.064}, pmid = {22353756}, issn = {1872-6240}, mesh = {Adaptor Proteins, Signal Transducing/*metabolism ; Age Factors ; Amyotrophic Lateral Sclerosis/genetics/*pathology ; Animals ; Cytoskeletal Proteins/*metabolism ; Disease Models, Animal ; Gene Expression Regulation/genetics ; Glutathione/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Heat-Shock Proteins/*metabolism ; Heme Oxygenase-1/metabolism ; Humans ; Kelch-Like ECH-Associated Protein 1 ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/*metabolism ; NF-E2-Related Factor 2/*metabolism ; Neuroglia/metabolism/pathology ; Spinal Cord/*pathology ; Superoxide Dismutase/genetics ; Thioredoxins/metabolism ; }, abstract = {The Kelch-like ECH-associated protein 1 (Keap1)/Nuclear erythroid 2-related factor 2 (Nrf2) system is the major cellular defense mechanism under oxidative stress, but the role in motor neuron degeneration under amyotrophic lateral sclerosis (ALS) pathology has not yet been fully elucidated. Here we examined temporal and spatial changes of Keap1, Nrf2, and their downstream stress response proteins heme oxgenase-1 (HO-1), glutathione, thioredoxin (TRX), and heat shock protein 70 (HSP70) throughout the course of motor neuron (MN) degeneration in the spinal cord of ALS model mice. Keap1 protein levels progressively decreased in the MN and anterior lumbar cord of ALS mice to 63% at early symptomatic 14 weeks and 58% at end symptomatic 18 weeks, while Nrf2 dramatically increased in the anterior lumbar cord with accumulation in the MN nucleus to 229% at 14 weeks and 471% at 18 weeks when glial like cells became also positive. In contrast, downstream stress response proteins such as HO-1, glutathione, TRX, and HSP70 showed only a small increase in MN with a significant increase to 149% to 280% in the number of glial-like cells after symptomatic 14 weeks. Our present observation suggests that MN selectively lost inductions of these important downstream protective proteins without regard to the Keap1/Nrf2 system activation, which could be a pivotal mechanism of neurodegenerative processes of ALS.}, }
@article {pmid22384095, year = {2012}, author = {Gregory, JM and Barros, TP and Meehan, S and Dobson, CM and Luheshi, LM}, title = {The aggregation and neurotoxicity of TDP-43 and its ALS-associated 25 kDa fragment are differentially affected by molecular chaperones in Drosophila.}, journal = {PloS one}, volume = {7}, number = {2}, pages = {e31899}, pmid = {22384095}, issn = {1932-6203}, support = {089703/WT_/Wellcome Trust/United Kingdom ; G0700990/MRC_/Medical Research Council/United Kingdom ; MC_G1000734/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/*metabolism ; Animals ; Animals, Genetically Modified ; Benzoquinones/pharmacology ; Brain/metabolism ; DNA-Binding Proteins/chemistry/*metabolism ; Drosophila Proteins/chemistry/*metabolism ; Drosophila melanogaster/*metabolism ; *Gene Expression Regulation ; Heat-Shock Proteins/*metabolism ; Heat-Shock Proteins, Small/chemistry/*metabolism ; Lactams, Macrocyclic/pharmacology ; Microscopy, Electron, Scanning/methods ; Models, Biological ; Models, Genetic ; Peptide Fragments/chemistry/*metabolism ; Phenotype ; Transgenes ; }, abstract = {Almost all cases of sporadic amyotrophic lateral sclerosis (ALS), and some cases of the familial form, are characterised by the deposition of TDP-43, a member of a family of heteronuclear ribonucleoproteins (hnRNP). Although protein misfolding and deposition is thought to be a causative feature of many of the most prevalent neurodegenerative diseases, a link between TDP-43 aggregation and the dysfunction of motor neurons has yet to be established, despite many correlative neuropathological studies. We have investigated this relationship in the present study by probing the effect of altering TDP-43 aggregation behaviour in vivo by modulating the levels of molecular chaperones in a Drosophila model. More specifically, we quantify the effect of either pharmacological upregulation of the heat shock response or specific genetic upregulation of a small heat shock protein, CG14207, on the neurotoxicity of both TDP-43 and of its disease associated 25 kDa fragment (TDP-25) in a Drosophila model. Inhibition of the aggregation of TDP-43 by either method results in a partial reduction of its neurotoxic effects on both photoreceptor and motor neurons, whereas inhibition of the aggregation of TDP-25 results not only in a complete suppression of its toxicity but also its clearance from the brain in both neuronal subtypes studied. The results demonstrate, therefore, that aggregation plays a crucial role in mediating the neurotoxic effects of both full length and truncated TDP-43, and furthermore reveal that the in vivo propensity of these two proteins to aggregate and their susceptibility to molecular chaperone mediated clearance are quite distinct.}, }
@article {pmid22476656, year = {2012}, author = {Banno, H and Katsuno, M and Suzuki, K and Tanaka, F and Sobue, G}, title = {Pathogenesis and molecular targeted therapy of spinal and bulbar muscular atrophy (SBMA).}, journal = {Cell and tissue research}, volume = {349}, number = {1}, pages = {313-320}, doi = {10.1007/s00441-012-1377-9}, pmid = {22476656}, issn = {1432-0878}, mesh = {Animals ; Bulbo-Spinal Atrophy, X-Linked/*drug therapy/*etiology/genetics ; Clinical Trials as Topic ; Humans ; Leuprolide/therapeutic use ; *Molecular Targeted Therapy ; }, abstract = {Spinal and bulbar muscular atrophy (SBMA), also known as Kennedy's disease, is an adult-onset, X-linked motor neuron disease characterized by muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. SBMA is caused by the expansion of a CAG triplet repeat, encoding a polyglutamine tract within the first exon of the androgen receptor (AR) gene. The histopathological finding in SBMA is the loss of lower motor neurons in the anterior horn of the spinal cord as well as in the brainstem motor nuclei. There is no established disease-modifying therapy for SBMA. Animal studies have revealed that the pathogenesis of SBMA depends on the level of serum testosterone, and that androgen deprivation mitigates neurodegeneration through inhibition of nuclear accumulation and/or stabilization of the pathogenic AR. Heat shock proteins, the ubiquitin-proteasome system and transcriptional regulation are also potential targets for development of therapy for SBMA. Among these therapeutic approaches, the luteinizing hormone-releasing hormone analogue, leuprorelin, prevents nuclear translocation of aberrant AR proteins, resulting in a significant improvement of disease phenotype in a mouse model of SBMA. In a phase 2 clinical trial of leuprorelin, the patients treated with this drug exhibited decreased mutant AR accumulation in scrotal skin biopsy. Phase 3 clinical trial showed the possibility that leuprorelin treatment is associated with improved swallowing function particularly in patients with a disease duration less than 10 years. These observations suggest that pharmacological inhibition of the toxic accumulation of mutant AR is a potential therapy for SBMA.}, }
@article {pmid22521462, year = {2012}, author = {Srivastava, AK and Renusch, SR and Naiman, NE and Gu, S and Sneh, A and Arnold, WD and Sahenk, Z and Kolb, SJ}, title = {Mutant HSPB1 overexpression in neurons is sufficient to cause age-related motor neuronopathy in mice.}, journal = {Neurobiology of disease}, volume = {47}, number = {2}, pages = {163-173}, pmid = {22521462}, issn = {1095-953X}, support = {K08 NS067282/NS/NINDS NIH HHS/United States ; K08NS067282/NS/NINDS NIH HHS/United States ; }, mesh = {Aging/*metabolism/pathology ; Animals ; Axons/pathology ; Charcot-Marie-Tooth Disease/*metabolism/pathology ; *Gene Expression Regulation ; HSP27 Heat-Shock Proteins/biosynthesis/*genetics ; Heat-Shock Proteins ; Humans ; Mice ; Mice, Transgenic ; Molecular Chaperones ; Motor Neuron Disease/metabolism/pathology ; Motor Neurons/*metabolism/pathology ; Mutation/*physiology ; Random Allocation ; }, abstract = {The small heat shock protein HSPB1 is a multifunctional, α-crystallin-based protein that has been shown to be neuroprotective in animal models of motor neuron disease and peripheral nerve injury. Missense mutations in HSPB1 result in axonal Charcot-Marie-Tooth disease with minimal sensory involvement (CMT2F) and distal hereditary motor neuropathy type 2 (dHMN-II). These disorders are characterized by a selective loss of motor axons in peripheral nerve resulting in distal muscle weakness and often severe disability. To investigate the pathogenic mechanisms of HSPB1 mutations in motor neurons in vivo, we have developed and characterized transgenic PrP-HSPB1 and PrP-HSPB1(R136W) mice. These mice express the human HSPB1 protein throughout the nervous system including in axons of peripheral nerve. Although both mouse strains lacked obvious motor deficits, the PrP-HSPB1(R136W) mice developed an age-dependent motor axonopathy. Mutant mice showed axonal pathology in spinal cord and peripheral nerve with evidence of impaired neurofilament cytoskeleton, associated with organelle accumulation. Accompanying these findings, increases in the number of Schmidt-Lanterman incisures, as evidence of impaired axon-Schwann cell interactions, were present. These observations suggest that overexpression of HSPB1(R136W) in neurons is sufficient to cause pathological and electrophysiological changes in mice that are seen in patients with hereditary motor neuropathy.}, }
@article {pmid22591194, year = {2012}, author = {Kalmar, B and Edet-Amana, E and Greensmith, L}, title = {Treatment with a coinducer of the heat shock response delays muscle denervation in the SOD1-G93A mouse model of amyotrophic lateral sclerosis.}, journal = {Amyotrophic lateral sclerosis : official publication of the World Federation of Neurology Research Group on Motor Neuron Diseases}, volume = {13}, number = {4}, pages = {378-392}, doi = {10.3109/17482968.2012.660953}, pmid = {22591194}, issn = {1471-180X}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Acetylcholinesterase/*drug effects/metabolism ; Amyotrophic Lateral Sclerosis/*drug therapy/metabolism/pathology ; Animals ; Choline O-Acetyltransferase/*drug effects/metabolism ; Disease Models, Animal ; Disease Progression ; GPI-Linked Proteins/drug effects/metabolism ; HSP70 Heat-Shock Proteins/drug effects/metabolism ; Heat-Shock Response ; Hydroxylamines/*pharmacology ; Longitudinal Studies ; Mice ; Mice, Transgenic ; Motor Neurons/*drug effects/metabolism ; Muscle Fibers, Fast-Twitch/drug effects/metabolism ; Muscle Fibers, Slow-Twitch/drug effects/metabolism ; Muscle, Skeletal/drug effects/*innervation/metabolism ; Neuromuscular Junction/*drug effects/metabolism ; Succinate Dehydrogenase/drug effects/metabolism ; Superoxide Dismutase/genetics ; }, abstract = {We undertook a longitudinal study of the histological and biochemical changes at the neuromuscular junction (NMJ) in muscles of SOD1-G93A mice. We also assessed these functions in mice treated with a known heat shock protein inducer, arimoclomol. Tissue samples of treated and untreated mSOD mice were analysed for AChE and ChAT enzyme activities as markers of neuromuscular function. Sections of hindlimb muscles (TA, EDL and soleus) were also stained for succinate dehydrogenase and silver cholinesterase activities as well as for immunohistochemistry. Hsp70 levels were also measured from muscle samples using ELISA. Results showed that denervation and nerve sprouting were present at symptom onset in fast muscles, although slow muscles remained fully innervated. Cholinergic enzyme activities were reduced prior to denervation and declined further with disease progression. Reduction of endplate size, a slow to fast shift in muscle phenotype was also observed. Treatment with arimoclomol delayed the appearance of these changes, increased innervation, cholinergic enzyme activities and endplate size and reversed muscle fibre transformation. These beneficial effects of arimoclomol in muscles were accompanied by an increase in Hsp70 expression. In conclusion, our results indicate that pharmacological targeting of muscles at early stages of disease may be a successful strategy to ameliorate disease progression in ALS.}, }
@article {pmid22745655, year = {2012}, author = {Knippenberg, S and Thau, N and Dengler, R and Brinker, T and Petri, S}, title = {Intracerebroventricular injection of encapsulated human mesenchymal cells producing glucagon-like peptide 1 prolongs survival in a mouse model of ALS.}, journal = {PloS one}, volume = {7}, number = {6}, pages = {e36857}, pmid = {22745655}, issn = {1932-6203}, mesh = {Amyotrophic Lateral Sclerosis/*metabolism/*therapy ; Animals ; Female ; Glucagon-Like Peptide 1/genetics/*metabolism ; Humans ; Male ; Mesenchymal Stem Cells/*cytology/physiology ; Mice ; Mice, Transgenic ; Motor Neurons/metabolism ; Nitric Oxide Synthase Type I/genetics/metabolism ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1 ; }, abstract = {BACKGROUND: As pharmacological therapies have largely failed so far, stem cell therapy has recently come into the focus of ALS research. Neuroprotective potential was shown for several types of stem and progenitor cells, mainly due to release of trophic factors. In the present study, we assessed the effects of intracerebroventricular injection of glucagon-like peptide 1 (GLP-1) releasing mesenchymal stromal cells (MSC) in mutant SOD1 (G93A) transgenic mice.<br><br>To improve the neuroprotective effects of native MSC, they had been transfected with a plasmid vector encoding a GLP-1 fusion gene prior to the injection, as GLP-1 was shown to exhibit neuroprotective properties before. Cells were encapsulated and therefore protected against rejection. After intracerebroventricular injection of these GLP-1 MSC capsules in presymptomatic SOD1 (G93A) mice, we assessed possible protective effects by survival analysis, measurement of body weight, daily monitoring and evaluation of motor performance by rotarod and footprint analyses. Motor neuron numbers in the spinal cord as well as the amount of astrocytosis, microglial activation, heat shock response and neuronal nitric oxide synthase (nNOS) expression were analyzed by immunohistological methods. Treatment with GLP-1 producing MSC capsules significantly prolonged survival by 13 days, delayed symptom onset by 15 days and weight loss by 14 days and led to significant improvements in motor performance tests compared to vehicle treated controls. Histological data are mainly in favour of anti-inflammatory effects of GLP-1 producing MSC capsules with reduced detection of inflammatory markers and a significant heat shock protein increase.<br><br>CONCLUSION/SIGNIFICANCE: Intracerebroventricular injection of GLP-1 MSC capsules shows neuroprotective potential in the SOD1 (G93A) mouse model.}, }
@article {pmid22865541, year = {2013}, author = {Chow, AM and Tang, DW and Hanif, A and Brown, IR}, title = {Induction of heat shock proteins in cerebral cortical cultures by celastrol.}, journal = {Cell stress &amp; chaperones}, volume = {18}, number = {2}, pages = {155-160}, pmid = {22865541}, issn = {1466-1268}, mesh = {Animals ; Blotting, Western ; Cells, Cultured ; Cerebral Cortex/cytology/*drug effects/metabolism ; HSP27 Heat-Shock Proteins/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Heat-Shock Proteins/*metabolism ; Heme Oxygenase-1/metabolism ; Pentacyclic Triterpenes ; Potassium Channel Blockers/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Temperature ; Triterpenes/*pharmacology ; }, abstract = {Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS) are 'protein misfolding disorders' of the mature nervous system that are characterized by the accumulation of protein aggregates and selective cell loss. Different brain regions are impacted, with Alzheimer's affecting cells in the cerebral cortex, Parkinson's targeting dopaminergic cells in the substantia nigra and ALS causing degeneration of cells in the spinal cord. These diseases differ widely in frequency in the human population. Alzheimer's is more frequent than Parkinson's and ALS. Heat shock proteins (Hsps) are 'protein repair agents' that provide a line of defense against misfolded, aggregation-prone proteins. We have suggested that differing levels of constitutively expressed Hsps (Hsc70 and Hsp27) in neural cell populations confer a variable buffering capacity against 'protein misfolding disorders' that correlates with the relative frequencies of these neurodegenerative diseases. The high relative frequency of Alzheimer's may due to low levels of Hsc70 and Hsp27 in affected cell populations that results in a reduced defense capacity against protein misfolding. Here, we demonstrate that celastrol, but not classical heat shock treatment, is effective in inducing a set of neuroprotective Hsps in cultures derived from cerebral cortices, including Hsp70, Hsp27 and Hsp32. This set of Hsps is induced by celastrol at 'days in vitro' (DIV) 13 when cultured cortical cells reached maturity. The inducibility of a set of neuroprotective Hsps in mature cortical cultures at DIV13 suggests that celastrol is a potential agent to counter Alzheimer's disease, a neurodegenerative 'protein misfolding disorder' of the adult brain that targets cells in the cerebral cortex.}, }
@article {pmid22900172, year = {2012}, author = {Gifondorwa, DJ and Jimenz-Moreno, R and Hayes, CD and Rouhani, H and Robinson, MB and Strupe, JL and Caress, J and Milligan, C}, title = {Administration of Recombinant Heat Shock Protein 70 Delays Peripheral Muscle Denervation in the SOD1(G93A) Mouse Model of Amyotrophic Lateral Sclerosis.}, journal = {Neurology research international}, volume = {2012}, number = {}, pages = {170426}, pmid = {22900172}, issn = {2090-1860}, support = {R01 NS036081/NS/NINDS NIH HHS/United States ; R01 NS046615/NS/NINDS NIH HHS/United States ; R01 NS069212/NS/NINDS NIH HHS/United States ; R29 NS036081/NS/NINDS NIH HHS/United States ; }, abstract = {A prominent clinical feature of ALS is muscle weakness due to dysfunction, denervation and degeneration of motoneurons (MNs). While MN degeneration is a late stage event in the ALS mouse model, muscle denervation occurs significantly earlier in the disease. Strategies to prevent this early denervation may improve quality of life by maintaining muscle control and slowing disease progression. The precise cause of MN dysfunction and denervation is not known, but several mechanisms have been proposed that involve potentially toxic intra- and extracellular changes. Many cells confront these changes by mounting a stress response that includes increased expression of heat shock protein 70 (Hsp70). MNs do not upregulate Hsp70, and this may result in a potentially increased vulnerability. We previously reported that recombinant human hsp70 (rhHsp70) injections delayed symptom onset and increased lifespan in SOD1(G93A) mice. The exogenous rhHsp70 was localized to the muscle and not to spinal cord or brain suggesting it modulates peripheral pathophysiology. In the current study, we focused on earlier administration of Hsp70 and its effect on initial muscle denervation. Injections of the protein appeared to arrest denervation with preserved large myelinated peripheral axons, and reduced glial activation.}, }
@article {pmid22920902, year = {2013}, author = {Crul, T and Toth, N and Piotto, S and Literati-Nagy, P and Tory, K and Haldimann, P and Kalmar, B and Greensmith, L and Torok, Z and Balogh, G and Gombos, I and Campana, F and Concilio, S and Gallyas, F and Nagy, G and Berente, Z and Gungor, B and Peter, M and Glatz, A and Hunya, A and Literati-Nagy, Z and Vigh, L and Hoogstra-Berends, F and Heeres, A and Kuipers, I and Loen, L and Seerden, JP and Zhang, D and Meijering, RA and Henning, RH and Brundel, BJ and Kampinga, HH and Koranyi, L and Szilvassy, Z and Mandl, J and Sumegi, B and Febbraio, MA and Horvath, I and Hooper, PL and Vigh, L}, title = {Hydroximic acid derivatives: pleiotropic HSP co-inducers restoring homeostasis and robustness.}, journal = {Current pharmaceutical design}, volume = {19}, number = {3}, pages = {309-346}, doi = {10.2174/138161213804143716}, pmid = {22920902}, issn = {1873-4286}, support = {MR/K000608/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Genetic Pleiotropy/*physiology ; Heat-Shock Proteins/*biosynthesis/chemistry/genetics ; Heat-Shock Response/*physiology ; Homeostasis/*physiology ; Humans ; Membrane Lipids/chemistry/genetics/metabolism ; Oximes/chemistry/*metabolism ; }, abstract = {According to the "membrane sensor" hypothesis, the membrane's physical properties and microdomain organization play an initiating role in the heat shock response. Clinical conditions such as cancer, diabetes and neurodegenerative diseases are all coupled with specific changes in the physical state and lipid composition of cellular membranes and characterized by altered heat shock protein levels in cells suggesting that these "membrane defects" can cause suboptimal hsp-gene expression. Such observations provide a new rationale for the introduction of novel, heat shock protein modulating drug candidates. Intercalating compounds can be used to alter membrane properties and by doing so normalize dysregulated expression of heat shock proteins, resulting in a beneficial therapeutic effect for reversing the pathological impact of disease. The membrane (and lipid) interacting hydroximic acid (HA) derivatives discussed in this review physiologically restore the heat shock protein stress response, creating a new class of "membrane-lipid therapy" pharmaceuticals. The diseases that HA derivatives potentially target are diverse and include, among others, insulin resistance and diabetes, neuropathy, atrial fibrillation, and amyotrophic lateral sclerosis. At a molecular level HA derivatives are broad spectrum, multi-target compounds as they fluidize yet stabilize membranes and remodel their lipid rafts while otherwise acting as PARP inhibitors. The HA derivatives have the potential to ameliorate disparate conditions, whether of acute or chronic nature. Many of these diseases presently are either untreatable or inadequately treated with currently available pharmaceuticals. Ultimately, the HA derivatives promise to play a major role in future pharmacotherapy.}, }
@article {pmid22926189, year = {2013}, author = {Brotherton, TE and Li, Y and Glass, JD}, title = {Cellular toxicity of mutant SOD1 protein is linked to an easily soluble, non-aggregated form in vitro.}, journal = {Neurobiology of disease}, volume = {49}, number = {}, pages = {49-56}, pmid = {22926189}, issn = {1095-953X}, support = {P30 NS055077/NS/NINDS NIH HHS/United States ; P50 AG025688/AG/NIA NIH HHS/United States ; }, mesh = {Animals ; CHO Cells ; Cell Survival/drug effects/physiology ; Cricetulus ; HSP70 Heat-Shock Proteins/metabolism ; Humans ; Models, Molecular ; Mutant Proteins/genetics/*metabolism ; Mutation ; Proteasome Endopeptidase Complex/metabolism ; Proteasome Inhibitors/pharmacology ; Protein Folding ; Solubility ; Superoxide Dismutase/genetics/*metabolism ; Superoxide Dismutase-1 ; Transfection ; }, abstract = {Mutations in superoxide dismutase 1 (SOD1) are found in approximately 20% of patients with familial amyotrophic lateral sclerosis. The propensity of mutant SOD1 to form aggregates in pathologically affected cells (i.e. motor neurons) has implicated these poorly soluble protein aggregates and/or their misfolded soluble precursors as being instrumental to the disease process. We investigated the relative solubility and toxicity of four different mutant SOD1 proteins in a cell-based model system and demonstrate that the mutant, misfolded SOD1 proteins that are the most soluble are also the most toxic. This toxicity was ameliorated by upregulating heat-shock protein chaperones in order to refold the soluble, misfolded protein, regardless of the presence of poorly soluble SOD1. We further demonstrate that increasing the solubility of a SOD1 mutant protein that is both poorly soluble and non-toxic, as compared to other mutant proteins, resulted in remarkably increased toxicity of the mutant SOD1. Again, this increased toxicity was attenuated by upregulating heat-shock protein chaperones in order to refold the soluble, misfolded proteins. These findings implicate easily soluble, misfolded SOD1 as being toxic to the cell and support the hypothesis that reducing solubility of mutant SOD1 proteins through aggregation may occur as a self-protective response in the cell.}, }
@article {pmid22993064, year = {2013}, author = {Yerbury, JJ and Gower, D and Vanags, L and Roberts, K and Lee, JA and Ecroyd, H}, title = {The small heat shock proteins αB-crystallin and Hsp27 suppress SOD1 aggregation in vitro.}, journal = {Cell stress &amp; chaperones}, volume = {18}, number = {2}, pages = {251-257}, pmid = {22993064}, issn = {1466-1268}, mesh = {Amino Acid Substitution ; Benzothiazoles ; HSP27 Heat-Shock Proteins/*metabolism ; Humans ; Spectrometry, Fluorescence ; Superoxide Dismutase/chemistry/genetics/*metabolism ; Superoxide Dismutase-1 ; Thiazoles/chemistry/metabolism ; alpha-Crystallin B Chain/*metabolism ; }, abstract = {Amyotrophic lateral sclerosis is a devastating neurodegenerative disease. The mechanism that underlies amyotrophic lateral sclerosis (ALS) pathology remains unclear, but protein inclusions are associated with all forms of the disease. Apart from pathogenic proteins, such as TDP-43 and SOD1, other proteins are associated with ALS inclusions including small heat shock proteins. However, whether small heat shock proteins have a direct effect on SOD1 aggregation remains unknown. In this study, we have examined the ability of small heat shock proteins αB-crystallin and Hsp27 to inhibit the aggregation of SOD1 in vitro. We show that these chaperone proteins suppress the increase in thioflavin T fluorescence associated with SOD1 aggregation, primarily through inhibiting aggregate growth, not the lag phase in which nuclei are formed. αB-crystallin forms high molecular mass complexes with SOD1 and binds directly to SOD1 aggregates. Our data are consistent with an overload of proteostasis systems being associated with pathology in ALS.}, }
@article {pmid23000195, year = {2012}, author = {Han, S and Choi, JR and Soon Shin, K and Kang, SJ}, title = {Resveratrol upregulated heat shock proteins and extended the survival of G93A-SOD1 mice.}, journal = {Brain research}, volume = {1483}, number = {}, pages = {112-117}, doi = {10.1016/j.brainres.2012.09.022}, pmid = {23000195}, issn = {1872-6240}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/genetics/*mortality ; Animals ; Antioxidants/*therapeutic use ; DNA-Binding Proteins/metabolism ; Disease Models, Animal ; Glial Fibrillary Acidic Protein/metabolism ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Heat Shock Transcription Factors ; Heat-Shock Proteins/*metabolism ; Humans ; Macrophage-1 Antigen/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Molecular Chaperones ; Motor Neurons/drug effects ; Neoplasm Proteins/metabolism ; Resveratrol ; Spinal Cord/pathology ; Stilbenes/*therapeutic use ; Superoxide Dismutase/genetics ; Survival Analysis ; Transcription Factors/metabolism ; Up-Regulation/*drug effects/genetics ; }, abstract = {In the present study, we investigated whether resveratrol, a SIRT1 activator, can suppress the motor neuron degeneration in a transgenic mouse model of amyotrophic lateral sclerosis. Chronic intraperitoneal injection of resveratrol delayed the disease onset and extended survival of the transgenic mice overexpressing G93A-SOD1. The number of surviving motor neurons increased in the resveratrol-injected G93A mice. Importantly, the levels of Hsp25 and Hsp70 were elevated while the level of heat shock factor 1 (HSF1) acetylation decreased in the spinal cords of the resveratrol-injected G93A mice. Our data suggest that resveratrol may protect motor neurons from the mutant SOD1-induced neurotoxicity by promoting SIRT1-mediated deacetylation of HSF1 and subsequent upregulation of Hsps.}, }
@article {pmid23063459, year = {2012}, author = {Mimoto, T and Morimoto, N and Miyazaki, K and Kurata, T and Sato, K and Ikeda, Y and Abe, K}, title = {Expression of heat shock transcription factor 1 and its downstream target protein T-cell death associated gene 51 in the spinal cord of a mouse model of amyotrophic lateral sclerosis.}, journal = {Brain research}, volume = {1488}, number = {}, pages = {123-131}, doi = {10.1016/j.brainres.2012.10.012}, pmid = {23063459}, issn = {1872-6240}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/metabolism/pathology ; Animals ; DNA-Binding Proteins/*genetics/metabolism ; Disease Models, Animal ; Disease Progression ; Female ; Heat Shock Transcription Factors ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/pathology/physiology ; Nerve Degeneration/genetics/metabolism/pathology ; Spinal Cord/pathology/*physiology ; Stress, Physiological/physiology ; T-Lymphocytes/*physiology ; Transcription Factors/*genetics/metabolism ; }, abstract = {Heat shock transcription factor 1 (HSF1) modulates the expression of the cell survival heat shock protein 70 (HSP70) and the cell death T-cell death associated gene 51 (TDAG51) in response to heat shock and various other cell stressors. We previously reported an increase in HSP70 in glial cells of the spinal anterior horn. Here we examined the temporal and spatial changes of HSF1 and TDAG51 expression over the course of motor neuron degeneration in the spinal cord of a mouse model of ALS (G93A-SOD1 Tg mice). The number of glial-like cells expressing HSF1 increased in G93A-SOD1 Tg mice at both early symptomatic (14 weeks) and end stages of disease (18 weeks), while the number of spinal neurons expressing HSF1 decreased. The total level of HSF1 in the anterior lumbar spinal cord was significantly decreased in G93A-SOD1 Tg mice at the end stage of disease. In contrast to HSF1, the level of TDAG51 in the anterior lumbar spinal cord was significantly increased in G93A-SOD1 Tg mice at the end stage of disease. Moreover, TDAG51 progressively increased in glial-like cells in the anterior lumbar spinal cord of G93A-SOD1 Tg mice from the early symptomatic stage, while decreasing in spinal neurons. Taken together, our results suggest that the balance between the cell survival and death signals mediated by HSP70 and TDAG51, respectively, may be disturbed by the altered expression of HSF1 during the progression of disease in this ALS model.}, }
@article {pmid23180277, year = {2013}, author = {Jung, MK and Kim, KY and Lee, NY and Kang, YS and Hwang, YJ and Kim, Y and Sung, JJ and McKee, A and Kowall, N and Lee, J and Ryu, H}, title = {Expression of taurine transporter (TauT) is modulated by heat shock factor 1 (HSF1) in motor neurons of ALS.}, journal = {Molecular neurobiology}, volume = {47}, number = {2}, pages = {699-710}, pmid = {23180277}, issn = {1559-1182}, support = {P30 AG013846/AG/NIA NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism ; Animals ; Base Sequence ; Cells, Cultured ; DNA-Binding Proteins/*physiology ; *Gene Expression Regulation ; Heat Shock Transcription Factors ; Male ; Membrane Glycoproteins/*biosynthesis/genetics/*metabolism ; Membrane Transport Proteins/*biosynthesis/genetics/*metabolism ; Mice ; Mice, Transgenic ; Molecular Sequence Data ; Motor Neurons/metabolism/*physiology ; Transcription Factors/*physiology ; Taurine Transporters ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurological disorder characterized by progressive paralysis caused by the degeneration of motor neurons throughout the central nervous system. Mutations of the free radical scavenging enzyme Cu/Zn superoxide dismutase 1 (SOD1) are a cause of familial ALS. In the present study, we demonstrated an age-dependent increase in taurine transporter (TauT) immunoreactivity in spinal cord motor neurons of ALS transgenic mice (mutant SOD1 (G93A)) and a similar increase in TauT in spinal motor neurons of patients with ALS. Chromatin immunoprecipitation analysis verified that heat shock factor 1 (HSF1) preferentially occupies the HSF1 binding element in the promoter of TauT under oxidative stress conditions. Knockdown of HSF1 by small interfering RNA reduced the transcriptional activity of TauT. Using [(3)H] taurine, we confirmed that an elevated expression of TauT directly contributes to increased taurine uptake in ALS motor neurons. In addition, we showed that taurine plays an antioxidant role and may prevent motor neuron loss due to oxidative stress in ALS. Our findings suggest that HSF1-induced TauT expression partially protects motor neurons by compensating for constitutive oxidative stress, which is thought to be a key mechanism contributing to the pathogenesis of ALS. Taken together, our results suggest that TauT is a novel pathological marker for stressed motor neurons in ALS and that modulation of TauT and taurine may slow neuronal degeneration in ALS.}, }
@article {pmid23281025, year = {2013}, author = {McGown, A and McDearmid, JR and Panagiotaki, N and Tong, H and Al Mashhadi, S and Redhead, N and Lyon, AN and Beattie, CE and Shaw, PJ and Ramesh, TM}, title = {Early interneuron dysfunction in ALS: insights from a mutant sod1 zebrafish model.}, journal = {Annals of neurology}, volume = {73}, number = {2}, pages = {246-258}, pmid = {23281025}, issn = {1531-8249}, support = {R01NS050414/NS/NINDS NIH HHS/United States ; BB/F01516X/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; RAMESH/APR11/808-791/MNDA_/Motor Neurone Disease Association/United Kingdom ; P30 NS045758/NS/NINDS NIH HHS/United States ; R01 NS050414/NS/NINDS NIH HHS/United States ; G0700091/MRC_/Medical Research Council/United Kingdom ; P30NS045758/NS/NINDS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/drug therapy/genetics/pathology/*physiopathology ; Animals ; Animals, Genetically Modified ; Apomorphine/pharmacology ; *Disease Models, Animal ; Dopamine Agonists/pharmacology ; Genes, Reporter ; Glycine/physiology ; HSP72 Heat-Shock Proteins/genetics ; Humans ; Interneurons/drug effects/pathology/*physiology ; Mice ; Motor Neurons/drug effects/pathology/physiology ; Muscle, Skeletal/innervation ; NF-E2-Related Factor 2/metabolism ; Neuromuscular Junction/pathology/physiopathology ; Neuroprotective Agents ; Patch-Clamp Techniques ; Riluzole/pharmacology ; Stress, Physiological/drug effects/physiology ; Superoxide Dismutase/*genetics/metabolism ; Superoxide Dismutase-1 ; *Zebrafish ; Zebrafish Proteins/metabolism ; }, abstract = {OBJECTIVE: To determine, when, how, and which neurons initiate the onset of pathophysiology in amyotrophic lateral sclerosis (ALS) using a transgenic mutant sod1 zebrafish model and identify neuroprotective drugs.<br><br>METHODS: Proteinopathies such as ALS involve mutant proteins that misfold and activate the heat shock stress response (HSR). The HSR is indicative of neuronal stress, and we used a fluorescent hsp70-DsRed reporter in our transgenic zebrafish to track neuronal stress and to measure functional changes in neurons and muscle over the course of the disease.<br><br>RESULTS: We show that mutant sod1 fish first exhibited the HSR in glycinergic interneurons at 24 hours postfertilization (hpf). By 96 hpf, we observed a significant reduction in spontaneous glycinergic currents induced in spinal motor neurons. The loss of inhibition was followed by increased stress in the motor neurons of symptomatic adults and concurrent morphological changes at the neuromuscular junction (NMJ) indicative of denervation. Riluzole, the only approved ALS drug and apomorphine, an NRF2 activator, reduced the observed early neuronal stress response.<br><br>INTERPRETATION: The earliest event in the pathophysiology of ALS in the mutant sod1 zebrafish model involves neuronal stress in inhibitory interneurons, resulting from mutant Sod1 expression. This is followed by a reduction in inhibitory input to motor neurons. The loss of inhibitory input may contribute to the later development of neuronal stress in motor neurons and concurrent inability to maintain the NMJ. Riluzole, the approved drug for use in ALS, modulates neuronal stress in interneurons, indicating a novel mechanism of riluzole action.}, }
@article {pmid23393146, year = {2013}, author = {Malik, B and Nirmalananthan, N and Gray, AL and La Spada, AR and Hanna, MG and Greensmith, L}, title = {Co-induction of the heat shock response ameliorates disease progression in a mouse model of human spinal and bulbar muscular atrophy: implications for therapy.}, journal = {Brain : a journal of neurology}, volume = {136}, number = {Pt 3}, pages = {926-943}, pmid = {23393146}, issn = {1460-2156}, support = {G0601943/MRC_/Medical Research Council/United Kingdom ; MR/K000608/1/MRC_/Medical Research Council/United Kingdom ; R01 NS041648/NS/NINDS NIH HHS/United States ; R01 NS41648/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Blotting, Western ; Disease Models, Animal ; Disease Progression ; Heat-Shock Proteins/*metabolism ; Heat-Shock Response/*drug effects/physiology ; Hydroxylamines/*pharmacology ; Male ; Mice ; Muscular Disorders, Atrophic/*metabolism ; Neuroprotective Agents/*pharmacology ; Real-Time Polymerase Chain Reaction ; }, abstract = {Spinal and bulbar muscular atrophy, also known as Kennedy's disease, is an adult-onset hereditary neurodegenerative disorder caused by an expansion of the polyglutamine repeat in the first exon in the androgen receptor gene. Pathologically, the disease is defined by selective loss of spinal and bulbar motor neurons causing bulbar, facial and limb weakness. Although the precise disease pathophysiology is largely unknown, it appears to be related to abnormal accumulation of the pathogenic androgen receptor protein within the nucleus, leading to disruption of cellular processes. Using a mouse model of spinal and bulbar muscular atrophy that exhibits many of the characteristic features of the human disease, in vivo physiological assessment of muscle function revealed that mice with the pathogenic expansion of the androgen receptor develop a motor deficit characterized by a reduction in muscle force, abnormal muscle contractile characteristics, loss of functional motor units and motor neuron degeneration. We have previously shown that treatment with arimoclomol, a co-inducer of the heat shock stress response, delays disease progression in the mutant superoxide dismutase 1 mouse model of amyotrophic lateral sclerosis, a fatal motor neuron disease. We therefore evaluated the therapeutic potential of arimoclomol in mice with spinal and bulbar muscular atrophy. Arimoclomol was administered orally, in drinking water, from symptom onset and the effects established at 18 months of age, a late stage of disease. Arimoclomol significantly improved hindlimb muscle force and contractile characteristics, rescued motor units and, importantly, improved motor neuron survival and upregulated the expression of the vascular endothelial growth factor which possess neurotrophic activity. These results provide evidence that upregulation of the heat shock response by treatment with arimoclomol may have therapeutic potential in the treatment of spinal and bulbar muscular atrophy and may also be a possible approach for the treatment of other neurodegenerative diseases.}, }
@article {pmid23487751, year = {2013}, author = {Franco, MC and Ye, Y and Refakis, CA and Feldman, JL and Stokes, AL and Basso, M and Melero Fernández de Mera, RM and Sparrow, NA and Calingasan, NY and Kiaei, M and Rhoads, TW and Ma, TC and Grumet, M and Barnes, S and Beal, MF and Beckman, JS and Mehl, R and Estévez, AG}, title = {Nitration of Hsp90 induces cell death.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {110}, number = {12}, pages = {E1102-11}, pmid = {23487751}, issn = {1091-6490}, support = {NS36761/NS/NINDS NIH HHS/United States ; R01 NS058628/NS/NINDS NIH HHS/United States ; NS058628/NS/NINDS NIH HHS/United States ; S10 RR006487/RR/NCRR NIH HHS/United States ; R01 NS036761/NS/NINDS NIH HHS/United States ; P30 CA13148/CA/NCI NIH HHS/United States ; RR13795/RR/NCRR NIH HHS/United States ; RR06487/RR/NCRR NIH HHS/United States ; NS42834/NS/NINDS NIH HHS/United States ; ES00210/ES/NIEHS NIH HHS/United States ; AT002034/AT/NCCIH NIH HHS/United States ; P30 CA013148/CA/NCI NIH HHS/United States ; R01 NS042834/NS/NINDS NIH HHS/United States ; P01 AT002034/AT/NCCIH NIH HHS/United States ; P30 ES000210/ES/NIEHS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/metabolism ; Animals ; Cell Death/*physiology ; Disease Models, Animal ; HSP90 Heat-Shock Proteins/*metabolism ; Humans ; Motor Neurons/metabolism/pathology ; Peroxynitrous Acid/*metabolism ; Protein Processing, Post-Translational/*physiology ; Rats ; Spinal Cord Injuries/metabolism/pathology ; Tyrosine/metabolism ; fas Receptor/metabolism ; }, abstract = {Oxidative stress is a widely recognized cause of cell death associated with neurodegeneration, inflammation, and aging. Tyrosine nitration in these conditions has been reported extensively, but whether tyrosine nitration is a marker or plays a role in the cell-death processes was unknown. Here, we show that nitration of a single tyrosine residue on a small proportion of 90-kDa heat-shock protein (Hsp90), is sufficient to induce motor neuron death by the P2X7 receptor-dependent activation of the Fas pathway. Nitrotyrosine at position 33 or 56 stimulates a toxic gain of function that turns Hsp90 into a toxic protein. Using an antibody that recognizes the nitrated Hsp90, we found immunoreactivity in motor neurons of patients with amyotrophic lateral sclerosis, in an animal model of amyotrophic lateral sclerosis, and after experimental spinal cord injury. Our findings reveal that cell death can be triggered by nitration of a single protein and highlight nitrated Hsp90 as a potential target for the development of effective therapies for a large number of pathologies.}, }
@article {pmid23587065, year = {2013}, author = {Bentmann, E and Haass, C and Dormann, D}, title = {Stress granules in neurodegeneration--lessons learnt from TAR DNA binding protein of 43 kDa and fused in sarcoma.}, journal = {The FEBS journal}, volume = {280}, number = {18}, pages = {4348-4370}, doi = {10.1111/febs.12287}, pmid = {23587065}, issn = {1742-4658}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/metabolism/pathology ; Biomarkers/metabolism ; Cytoplasmic Granules/*genetics/metabolism/pathology ; DNA-Binding Proteins/*genetics/metabolism ; Frontotemporal Lobar Degeneration/*genetics/metabolism/pathology ; Gene Expression Regulation ; Heat-Shock Proteins/genetics/metabolism ; Humans ; Protein Folding ; Protein Structure, Tertiary ; RNA, Messenger/genetics/metabolism ; RNA-Binding Protein FUS/*genetics/metabolism ; Signal Transduction ; Stress, Physiological ; }, abstract = {Stress granules (SGs) are cytoplasmic foci that rapidly form when cells are exposed to stress. They transiently store mRNAs encoding house-keeping proteins and allow the selective translation of stress-response proteins (e.g. heat shock proteins). Besides mRNA, SGs contain RNA-binding proteins, such as T cell internal antigen-1 and poly(A)-binding protein 1, which can serve as characteristic SG marker proteins. Recently, some of these SG marker proteins were found to label pathological TAR DNA binding protein of 43 kDa (TDP-43)- or fused in sarcoma (FUS)-positive cytoplasmic inclusions in patients with amyotrophic lateral sclerosis and frontotemporal lobar degeneration. In addition, protein aggregates in other neurodegenerative diseases (e.g. tau inclusions in Alzheimer's disease) show a co-localization with T cell internal antigen-1 as well. Moreover, several RNA-binding proteins that are commonly found in SGs have been genetically linked to neurodegeneration. This suggests that SGs might play an important role in the pathogenesis of these proteinopathies, either by acting as a seed for pathological inclusions, by mediating translational repression or by trapping essential RNA-binding proteins, or by a combination of these mechanisms. This minireview gives an overview of the general biology of SGs and highlights the recently identified connection of SGs with TDP-43, FUS and other proteins involved in neurodegenerative diseases. We propose that pathological inclusions containing RNA-binding proteins, such as TDP-43 and FUS, might arise from SGs and discuss how SGs might contribute to neurodegeneration via toxic gain or loss-of-function mechanisms.}, }
@article {pmid23595219, year = {2013}, author = {Zinkie, S and Gentil, BJ and Minotti, S and Durham, HD}, title = {Expression of the protein chaperone, clusterin, in spinal cord cells constitutively and following cellular stress, and upregulation by treatment with Hsp90 inhibitor.}, journal = {Cell stress &amp; chaperones}, volume = {18}, number = {6}, pages = {745-758}, pmid = {23595219}, issn = {1466-1268}, support = {MOP77743//Canadian Institutes of Health Research/Canada ; }, mesh = {Amino Acid Substitution ; Amyotrophic Lateral Sclerosis/metabolism/pathology ; Animals ; Astrocytes/cytology/metabolism ; Benzoquinones/*pharmacology ; Cells, Cultured ; Clusterin/*metabolism ; Disease Models, Animal ; Female ; HSP90 Heat-Shock Proteins/*antagonists &amp; inhibitors/metabolism ; Lactams, Macrocyclic/*pharmacology ; Male ; Mice ; Mice, Transgenic ; Motor Neurons/cytology/metabolism ; Spinal Cord/cytology/*metabolism ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1 ; Temperature ; Up-Regulation/*drug effects ; }, abstract = {Clusterin, a protein chaperone found at high levels in physiological fluids, is expressed in nervous tissue and upregulated in several neurological diseases. To assess relevance to amyotrophic lateral sclerosis (ALS) and other motor neuron disorders, clusterin expression was evaluated using long-term dissociated cultures of murine spinal cord and SOD1(G93A) transgenic mice, a model of familial ALS. Motor neurons and astrocytes constitutively expressed nuclear and cytoplasmic forms of clusterin, and secreted clusterin accumulated in culture media. Although clusterin can be stress inducible, heat shock failed to increase levels in these neural cell compartments despite robust upregulation of stress-inducible Hsp70 (HspA1) in non-neuronal cells. In common with HSPs, clusterin was upregulated by treatment with the Hsp90 inhibitor, geldanamycin, and thus could contribute to the neuroprotection previously identified for such compounds in disease models. Clusterin expression was not altered in cultured motor neurons expressing SOD1(G93A) by gene transfer or in presymptomatic SOD1(G93A) transgenic mice; however, clusterin immunolabeling was weakly increased in lumbar spinal cord of overtly symptomatic mice. More striking, mutant SOD1 inclusions, a pathological hallmark, were strongly labeled by anti-clusterin. Since secreted, as well as intracellular, mutant SOD1 contributes to toxicity, the extracellular chaperoning property of clusterin could be important for folding and clearance of SOD1 and other misfolded proteins in the extracellular space. Evaluation of chaperone-based therapies should include evaluation of clusterin as well as HSPs, using experimental models that replicate the control mechanisms operant in the cells and tissue of interest.}, }
@article {pmid23625794, year = {2013}, author = {Sama, RR and Ward, CL and Kaushansky, LJ and Lemay, N and Ishigaki, S and Urano, F and Bosco, DA}, title = {FUS/TLS assembles into stress granules and is a prosurvival factor during hyperosmolar stress.}, journal = {Journal of cellular physiology}, volume = {228}, number = {11}, pages = {2222-2231}, pmid = {23625794}, issn = {1097-4652}, support = {R01 NS078145/NS/NINDS NIH HHS/United States ; R01NS078145-01/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Cell Death/drug effects ; Cell Nucleus/drug effects/metabolism ; Cell Survival/drug effects ; Cytoplasmic Granules/drug effects/*metabolism ; HEK293 Cells ; HeLa Cells ; Humans ; Hypertonic Solutions/*pharmacology ; Methylation/drug effects ; Mice ; RNA-Binding Protein FUS/*metabolism ; Sorbitol/pharmacology/toxicity ; Stress, Physiological/*drug effects ; }, abstract = {FUsed in Sarcoma/Translocated in LipoSarcoma (FUS/TLS or FUS) has been linked to several biological processes involving DNA and RNA processing, and has been associated with multiple diseases, including myxoid liposarcoma and amyotrophic lateral sclerosis (ALS). ALS-associated mutations cause FUS to associate with stalled translational complexes called stress granules under conditions of stress. However, little is known regarding the normal role of endogenous (non-disease linked) FUS in cellular stress response. Here, we demonstrate that endogenous FUS exerts a robust response to hyperosmolar stress induced by sorbitol. Hyperosmolar stress causes an immediate re-distribution of nuclear FUS to the cytoplasm, where it incorporates into stress granules. The redistribution of FUS to the cytoplasm is modulated by methyltransferase activity, whereas the inhibition of methyltransferase activity does not affect the incorporation of FUS into stress granules. The response to hyperosmolar stress is specific, since endogenous FUS does not redistribute to the cytoplasm in response to sodium arsenite, hydrogen peroxide, thapsigargin, or heat shock, all of which induce stress granule assembly. Intriguingly, cells with reduced expression of FUS exhibit a loss of cell viability in response to sorbitol, indicating a prosurvival role for endogenous FUS in the cellular response to hyperosmolar stress.}, }
@article {pmid23639787, year = {2013}, author = {Hwang, CS and Liu, GT and Chang, MD and Liao, IL and Chang, HT}, title = {Elevated serum autoantibody against high mobility group box 1 as a potent surrogate biomarker for amyotrophic lateral sclerosis.}, journal = {Neurobiology of disease}, volume = {58}, number = {}, pages = {13-18}, doi = {10.1016/j.nbd.2013.04.013}, pmid = {23639787}, issn = {1095-953X}, mesh = {Adult ; Age Factors ; Aged ; Aged, 80 and over ; Alzheimer Disease/blood ; Amyotrophic Lateral Sclerosis/*blood/classification/surgery ; Analysis of Variance ; Autoantibodies/*blood ; Biomarkers/*blood ; Chaperonin 60/immunology ; Cohort Studies ; Disease Progression ; Female ; HMGB1 Protein/*immunology ; HSP70 Heat-Shock Proteins/immunology ; Humans ; Male ; Middle Aged ; Mitochondrial Proteins/immunology ; Parkinson Disease/blood ; ROC Curve ; Tracheotomy/methods ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a complicate and progressive onset devastating neurodegenerative disease. Its pathogenic mechanisms remain unclear and there is no specific test for diagnosis. For years, researchers have been vigorously searching for biomarkers associated with ALS to assist clinical diagnosis and monitor disease progression. Some specific inflammatory processes in the central nervous system have been reported to participate in the pathogenesis of ALS. As high mobility group box 1 (HMGB1) is elevated in spinal cord tissues of patients with ALS, we hypothesized, therefore, that serum autoantibody against HMGB1 (HMGB1 autoAb) might represent an effective biomarker for ALS. Patients with ALS, Alzheimer's disease, Parkinson's disease, and healthy age-matched control subjects were recruited for this study. ALS group consisted of 61 subjects, the other groups each consisted of forty subjects. We generated a polyclonal antibody against HMGB1 and developed an ELISA-based methodology for screening serum samples of these subjects. All samples were coded for masked comparison. For statistic analyses, two-tailed Student's t-test, ANOVA, Bonferroni multiple comparison test, Spearman correlation, and receiver operating characteristic curve were applied. We discovered that the level of HMGB1 autoAb significantly increased in patients with ALS as compared with that of patients with Alzheimer's disease, Parkinson's disease, and healthy control subjects. The differences between all groups were robust even at the early stages of ALS progression. More importantly, higher HMGB1 autoAb level was found in more severe disease status with significant correlation. Our study demonstrates that serum HMGB1 autoAb may serve as a biomarker for the diagnosis of ALS and can be used to monitor disease progression.}, }
@article {pmid23711791, year = {2013}, author = {Harrison, IF and Dexter, DT}, title = {Epigenetic targeting of histone deacetylase: therapeutic potential in Parkinson's disease?.}, journal = {Pharmacology &amp; therapeutics}, volume = {140}, number = {1}, pages = {34-52}, doi = {10.1016/j.pharmthera.2013.05.010}, pmid = {23711791}, issn = {1879-016X}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Animals ; Epigenesis, Genetic ; Epigenomics ; Gene Expression ; Histone Deacetylase Inhibitors/pharmacology/therapeutic use ; Histone Deacetylases/*metabolism ; Humans ; Parkinson Disease/drug therapy/*genetics/*metabolism ; }, abstract = {Parkinson's disease (PD) is the most common movement disorder affecting more than 4million people worldwide. The primary motor symptoms of the disease are due to degeneration of dopaminergic nigrostriatal neurons. Dopamine replacement therapies have therefore revolutionised disease management by partially controlling these symptoms. However these drugs can produce debilitating side effects when used long term and do not protect degenerating neurons against death. Recent evidence has highlighted a pathological imbalance in PD between the acetylation and deacetylation of the histone proteins around which deoxyribonucleic acid (DNA) is coiled, in favour of excessive histone deacetylation. This mechanism of adding/removing acetyl groups to histone lysine residues is one of many epigenetic regulatory processes which control the expression of genes, many of which will be essential for neuronal survival. Hence, such epigenetic modifications may have a pathogenic role in PD. It has therefore been hypothesised that if this pathological imbalance can be corrected with the use of histone deacetylase inhibiting agents then neurodegeneration observed in PD can be ameliorated. This article will review the current literature with regard to epigenetic changes in PD and the use of histone deacetylase inhibitors (HDACIs) in PD: examining the evidence of the neuroprotective effects of numerous HDACIs in cellular and animal models of Parkinsonian cell death. Ultimately answering the question: does epigenetic targeting of histone deacetylases hold therapeutic potential in PD?}, }
@article {pmid23764168, year = {2013}, author = {Prell, T and Lautenschläger, J and Grosskreutz, J}, title = {Calcium-dependent protein folding in amyotrophic lateral sclerosis.}, journal = {Cell calcium}, volume = {54}, number = {2}, pages = {132-143}, doi = {10.1016/j.ceca.2013.05.007}, pmid = {23764168}, issn = {1532-1991}, mesh = {Amyotrophic Lateral Sclerosis/*physiopathology ; Animals ; Calcium/*physiology ; Disease Models, Animal ; Endoplasmic Reticulum/physiology ; Endoplasmic Reticulum Chaperone BiP ; Homeostasis/physiology ; Humans ; Mice ; *Protein Folding ; }, abstract = {Amyotrophic lateral sclerosis is a neurodegenerative disease characterized by a progressive loss of motor neurons. Although the etiology remains unclear, disturbances in Ca2+ homoeostasis and protein folding are essential features of neurodegeneration. The correct folding of proteins is managed by folding proteins, which are regulated by Ca2+ levels. Therefore, Ca(2+)-sensitive folding proteins represent an important link between disturbed Ca2+ handling and protein misfolding in amyotrophic lateral sclerosis. In the first part of this review, we focus on Ca2+ handling in the endoplasmic reticulum and mitochondria in terms of their roles in protein misfolding. In the second part, we draw attention to the main Ca(2+)-sensitive folding proteins that play a role in motor neuron degeneration such as calreticulin and calnexin, which are involved in the folding of glycosylated proteins. In addition, calmodulin and the Ca2+/calmodulin-dependent protein kinase are discussed as one correlation to oxidative stress. The heat shock protein endoplasmin is associated with the anti-apoptotic insulin-like growth factor pathway that is altered in amyotrophic lateral sclerosis. Grp78, which influences Ca2+ homeostasis in the intraluminal endoplasmic reticulum is upregulated in mice models and amyotrophic lateral sclerosis patients and constitutes a core component of the unfolded protein response. Lastly, the protein disulfide isomerase family is responsible for mediating oxidative protein folding in the endoplasmic reticulum.}, }
@article {pmid23827971, year = {2013}, author = {Salminen, A and Kaarniranta, K and Kauppinen, A and Ojala, J and Haapasalo, A and Soininen, H and Hiltunen, M}, title = {Impaired autophagy and APP processing in Alzheimer's disease: The potential role of Beclin 1 interactome.}, journal = {Progress in neurobiology}, volume = {106-107}, number = {}, pages = {33-54}, doi = {10.1016/j.pneurobio.2013.06.002}, pmid = {23827971}, issn = {1873-5118}, mesh = {Alzheimer Disease/genetics/*metabolism ; Amyloid beta-Protein Precursor/*metabolism ; Animals ; Apoptosis/physiology ; Apoptosis Regulatory Proteins/genetics/*metabolism ; Autophagy/genetics/*physiology ; Beclin-1 ; Humans ; Membrane Proteins/genetics/*metabolism ; Proteome/*metabolism ; }, abstract = {The accumulation of amyloid-β-containing neuritic plaques and intracellular tau protein tangles are key histopathological hallmarks of Alzheimer's disease (AD). This type of pathology clearly indicates that the mechanisms of neuronal housekeeping and protein quality control are compromised in AD. There is mounting evidence that the autophagosome-lysosomal degradation is impaired, which could disturb the processing of APP and provoke AD pathology. Beclin 1 is a molecular platform assembling an interactome with stimulating and suppressive components which regulate the initiation of the autophagosome formation. Recent studies have indicated that the expression Beclin 1 is reduced in AD brain. Moreover, the deficiency of Beclin 1 in cultured neurons and transgenic mice provokes the deposition of amyloid-β peptides whereas its overexpression reduces the accumulation of amyloid-β. There are several potential mechanisms, which could inhibit the function of Beclin 1 interactome and thus impair autophagy and promote AD pathology. The mechanisms include (i) reduction of Beclin 1 expression or its increased proteolytic cleavage by caspases, (ii) sequestration of Beclin 1 to non-functional locations, such as tau tangles, (iii) formation of inhibitory complexes between Beclin 1 and antiapoptotic Bcl-2 proteins or inflammasomes, (iv) interaction of Beclin 1 with inhibitory neurovirulent proteins, e.g. herpex simplex ICP34.5, or (v) inhibition of the Beclin 1/Vps34 complex through the activation of CDK1 and CDK5. We will shortly introduce the function of Beclin 1 interactome in autophagy and phagocytosis, review the recent evidence indicating that Beclin 1 regulates autophagy and APP processing in AD, and finally examine the potential mechanisms through which Beclin 1 dysfunction could be involved in the pathogenesis of AD.}, }
@article {pmid23886956, year = {2013}, author = {Wei, R and Bhattacharya, A and Hamilton, RT and Jernigan, AL and Chaudhuri, AR}, title = {Differential effects of mutant SOD1 on protein structure of skeletal muscle and spinal cord of familial amyotrophic lateral sclerosis: role of chaperone network.}, journal = {Biochemical and biophysical research communications}, volume = {438}, number = {1}, pages = {218-223}, doi = {10.1016/j.bbrc.2013.07.060}, pmid = {23886956}, issn = {1090-2104}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism ; Animals ; *Disease Models, Animal ; Heat-Shock Proteins/genetics/*metabolism ; Humans ; Male ; Mice ; Mice, Transgenic ; Molecular Chaperones/genetics/*metabolism ; Muscle, Skeletal/*metabolism ; Mutation/genetics ; Signal Transduction/genetics ; Species Specificity ; Spinal Cord/*metabolism ; Structure-Activity Relationship ; Superoxide Dismutase/genetics/*metabolism ; Tissue Distribution ; }, abstract = {Protein misfolding is considered to be a potential contributing factor for motor neuron and muscle loss in diseases like Amyotrophic lateral sclerosis (ALS). Several independent studies have demonstrated using over-expressed mutated Cu/Zn-superoxide dismutase (mSOD1) transgenic mouse models which mimic familial ALS (f-ALS), that both muscle and motor neurons undergo degeneration during disease progression. However, it is unknown whether protein conformation of skeletal muscle and spinal cord is equally or differentially affected by mSOD1-induced toxicity. It is also unclear whether heat shock proteins (Hsp's) differentially modulate skeletal muscle and spinal cord protein structure during ALS disease progression. We report three intriguing observations utilizing the f-ALS mouse model and cell-free in vitro system; (i) muscle proteins are equally sensitive to misfolding as spinal cord proteins despite the presence of low level of soluble and absence of insoluble G93A protein aggregate, unlike in spinal cord, (ii) Hsp's levels are lower in muscle compared to spinal cord at any stage of the disease, and (iii) G93ASOD1 enzyme-induced toxicity selectively affects muscle protein conformation over spinal cord proteins. Together, these findings strongly suggest that differential chaperone levels between skeletal muscle and spinal cord may be a critical determinant for G93A-induced protein misfolding in ALS.}, }
@article {pmid23962724, year = {2014}, author = {Udan-Johns, M and Bengoechea, R and Bell, S and Shao, J and Diamond, MI and True, HL and Weihl, CC and Baloh, RH}, title = {Prion-like nuclear aggregation of TDP-43 during heat shock is regulated by HSP40/70 chaperones.}, journal = {Human molecular genetics}, volume = {23}, number = {1}, pages = {157-170}, pmid = {23962724}, issn = {1460-2083}, support = {AG042095/AG/NIA NIH HHS/United States ; NS057105/NS/NINDS NIH HHS/United States ; R01 NS069669/NS/NINDS NIH HHS/United States ; AG031867/AG/NIA NIH HHS/United States ; NS069669/NS/NINDS NIH HHS/United States ; NS055980/NS/NINDS NIH HHS/United States ; R01 AR068797/AR/NIAMS NIH HHS/United States ; }, mesh = {Amino Acid Motifs ; Animals ; Brain/metabolism ; COS Cells ; Cell Nucleus/metabolism ; Chlorocebus aethiops ; Cytoplasm/metabolism ; DNA-Binding Proteins/*chemistry/*metabolism ; HEK293 Cells ; HSP40 Heat-Shock Proteins/*physiology ; HSP70 Heat-Shock Proteins/*physiology ; HeLa Cells ; Heat-Shock Response ; Heterogeneous Nuclear Ribonucleoprotein A1 ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B/*metabolism ; Humans ; Muscles/metabolism ; Prions/*chemistry/metabolism ; Protein Folding ; }, abstract = {TDP-43 aggregation in the cytoplasm or nucleus is a key feature of the pathology of amyotrophic lateral sclerosis and frontotemporal dementia and is observed in numerous other neurodegenerative diseases, including Alzheimer's disease. Despite this fact, the inciting events leading to TDP-43 aggregation remain unclear. We observed that endogenous TDP-43 undergoes reversible aggregation in the nucleus after the heat shock and that this behavior is mediated by the C-terminal prion domain. Substitution of the prion domain from TIA-1 or an authentic yeast prion domain from RNQ1 into TDP-43 can completely recapitulate heat shock-induced aggregation. TDP-43 is constitutively bound to members of the Hsp40/Hsp70 family, and we found that heat shock-induced TDP-43 aggregation is mediated by the availability of these chaperones interacting with the inherently disordered C-terminal prion domain. Finally, we observed that the aggregation of TDP-43 during heat shock led to decreased binding to hnRNPA1, and a change in TDP-43 RNA-binding partners suggesting that TDP-43 aggregation alters its function in response to misfolded protein stress. These findings indicate that TDP-43 shares properties with physiologic prions from yeast, in that self-aggregation is mediated by a Q/N-rich disordered domain, is modulated by chaperone proteins and leads to altered function of the protein. Furthermore, they indicate that TDP-43 aggregation is regulated by chaperone availability, explaining the recurrent observation of TDP-43 aggregates in degenerative diseases of both the brain and muscle where protein homeostasis is disrupted.}, }
@article {pmid23978556, year = {2014}, author = {Kalmar, B and Lu, CH and Greensmith, L}, title = {The role of heat shock proteins in Amyotrophic Lateral Sclerosis: The therapeutic potential of Arimoclomol.}, journal = {Pharmacology &amp; therapeutics}, volume = {141}, number = {1}, pages = {40-54}, doi = {10.1016/j.pharmthera.2013.08.003}, pmid = {23978556}, issn = {1879-016X}, support = {MR/K000608/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/*metabolism ; Animals ; Endoplasmic Reticulum Chaperone BiP ; Enzyme Induction/drug effects ; Heat-Shock Proteins/biosynthesis/*drug effects/*metabolism ; Heat-Shock Response/drug effects ; Humans ; Hydroxylamines/*pharmacology/*therapeutic use ; Models, Biological ; Molecular Targeted Therapy/methods ; Neuroprotective Agents/pharmacology/therapeutic use ; }, abstract = {Arimoclomol is a hydroxylamine derivative, a group of compounds which have unique properties as co-inducers of heat shock protein expression, but only under conditions of cellular stress. Arimoclomol has been found to be neuroprotective in a number of neurodegenerative disease models, including Amyotrophic Lateral Sclerosis (ALS), and in mutant Superoxide Dismutase 1 (SOD1) mice that model ALS, Arimoclomol rescues motor neurons, improves neuromuscular function and extends lifespan. The therapeutic potential of Arimoclomol is currently under investigation in a Phase II clinical trial for ALS patients with SOD1 mutations. In this review we summarize the evidence for the neuroprotective effects of enhanced heat shock protein expression by Arimoclomol and other inducers of the Heat Shock Response. ALS is a complex, multifactorial disease affecting a number of cell types and intracellular pathways. Cells and pathways affected by ALS pathology and which may be targeted by a heat shock protein-based therapy are also discussed in this review. For example, protein aggregation is a characteristic pathological feature of neurodegenerative diseases including ALS. Enhanced heat shock protein expression not only affects protein aggregation directly, but can also lead to more effective clearance of protein aggregates via the unfolded protein response, the proteasome-ubiquitin system or by autophagy. However, compounds such as Arimoclomol have effects beyond targeting protein mis-handling and can also affect additional pathological mechanisms such as oxidative stress. Therefore, by targeting multiple pathological mechanisms, compounds such as Arimoclomol may be particularly effective in the development of a disease-modifying therapy for ALS and other neurodegenerative disorders.}, }
@article {pmid24092395, year = {2014}, author = {Cha, JR and St Louis, KJ and Tradewell, ML and Gentil, BJ and Minotti, S and Jaffer, ZM and Chen, R and Rubenstein, AE and Durham, HD}, title = {A novel small molecule HSP90 inhibitor, NXD30001, differentially induces heat shock proteins in nervous tissue in culture and in vivo.}, journal = {Cell stress &amp; chaperones}, volume = {19}, number = {3}, pages = {421-435}, pmid = {24092395}, issn = {1466-1268}, support = {R21 NS066129/NS/NINDS NIH HHS/United States ; 1R21NS066129-01/NS/NINDS NIH HHS/United States ; MOP-77743/CAPMC/CIHR/Canada ; }, mesh = {Animals ; Calcium/metabolism ; Cell Survival/drug effects ; Cells, Cultured ; Ganglia, Spinal/drug effects/metabolism ; Green Fluorescent Proteins/metabolism ; HSP90 Heat-Shock Proteins/*antagonists &amp; inhibitors/metabolism ; Heat-Shock Proteins/*metabolism ; Homeostasis/drug effects ; Inclusion Bodies/metabolism ; Lactones/administration &amp; dosage/pharmacokinetics/*pharmacology ; Mice, Inbred C57BL ; Mice, Transgenic ; Mitochondrial Dynamics/drug effects ; Motor Neurons/drug effects/metabolism ; Nerve Tissue/drug effects/*metabolism ; Oximes/administration &amp; dosage/pharmacokinetics/*pharmacology ; Phosphorylation/drug effects ; Small Molecule Libraries/administration &amp; dosage/pharmacokinetics/*pharmacology ; Spinal Cord/drug effects/metabolism ; Superoxide Dismutase/metabolism ; Superoxide Dismutase-1 ; Tissue Culture Techniques ; }, abstract = {Heat shock proteins (HSPs) are attractive therapeutic targets for neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), characterized by aberrant formation of protein aggregates. Although motor neurons have a high threshold for activation of HSP genes, HSP90 inhibitors are effective inducers. This study evaluated NXD30001, a novel, small molecule HSP90 inhibitor based on the radicicol backbone, for its ability to induce neuronal HSPs and for efficacy in an experimental model of ALS based on mutations in superoxide-dismutase 1 (SOD1). In motor neurons of dissociated murine spinal cord cultures, NXD30001-induced expression of HSP70/HSPA1 (iHSP70) and its co-chaperone HSP40/DNAJ through activation of HSF1 and exhibited a protective profile against SOD1(G93A) similar to geldanamycin, but with less toxicity. Treatment prevented protein aggregation, mitochondrial fragmentation, and motor neuron death, important features of mutant SOD1 toxicity, but did not effectively prevent aberrant intracellular Ca(2+) accumulation. NXD30001 distributed to brain and spinal cord of wild-type and SOD1(G93A) transgenic mice following intraperitoneal injection; however, unlike in culture, in vivo levels of SOD1 were not reduced. NXD30001-induced expression of iHSP70 in skeletal and cardiac muscle and, to a lesser extent, in kidney, but not in liver, spinal cord, or brain, with either single or repeated administration. NXD30001 is a very useful experimental tool in culture, but these data point to the complex nature of HSP gene regulation in vivo and the necessity for early evaluation of the efficacy of novel HSP inducers in target tissues in vivo.}, }
@article {pmid24256261, year = {2013}, author = {Crippa, V and Galbiati, M and Boncoraglio, A and Rusmini, P and Onesto, E and Giorgetti, E and Cristofani, R and Zito, A and Poletti, A}, title = {Motoneuronal and muscle-selective removal of ALS-related misfolded proteins.}, journal = {Biochemical Society transactions}, volume = {41}, number = {6}, pages = {1598-1604}, doi = {10.1042/BST20130118}, pmid = {24256261}, issn = {1470-8752}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism/pathology ; Animals ; Humans ; Motor Neurons/*metabolism/pathology ; Muscles/*metabolism/pathology ; *Protein Folding ; Superoxide Dismutase/chemistry/*metabolism ; Superoxide Dismutase-1 ; }, abstract = {ALS (amyotrophic lateral sclerosis), a fatal motoneuron (motor neuron) disease, occurs in clinically indistinguishable sporadic (sALS) or familial (fALS) forms. Most fALS-related mutant proteins identified so far are prone to misfolding, and must be degraded in order to protect motoneurons from their toxicity. This process, mediated by molecular chaperones, requires proteasome or autophagic systems. Motoneurons are particularly sensitive to misfolded protein toxicity, but other cell types such as the muscle cells could also be affected. Muscle-restricted expression of the fALS protein mutSOD1 (mutant superoxide dismutase 1) induces muscle atrophy and motoneuron death. We found that several genes have an altered expression in muscles of transgenic ALS mice at different stages of disease. MyoD, myogenin, atrogin-1, TGFβ1 (transforming growth factor β1) and components of the cell response to proteotoxicity [HSPB8 (heat shock 22kDa protein 8), Bag3 (Bcl-2-associated athanogene 3) and p62] are all up-regulated by mutSOD1 in skeletal muscle. When we compared the potential mutSOD1 toxicity in motoneuron (NSC34) and muscle (C2C12) cells, we found that muscle ALS models possess much higher chymotryptic proteasome activity and autophagy power than motoneuron ALS models. As a result, mutSOD1 molecular behaviour was found to be very different. MutSOD1 clearance was found to be much higher in muscle than in motoneurons. MutSOD1 aggregated and impaired proteasomes only in motoneurons, which were particularly sensitive to superoxide-induced oxidative stress. Moreover, in muscle cells, mutSOD1 was found to be soluble even after proteasome inhibition. This effect could be associated with a higher mutSOD1 autophagic clearance. Therefore muscle cells seem to manage misfolded mutSOD1 more efficiently than motoneurons, thus mutSOD1 toxicity in muscle may not directly depend on aggregation.}, }
@article {pmid24256636, year = {2013}, author = {Lin, PY and Simon, SM and Koh, WK and Folorunso, O and Umbaugh, CS and Pierce, A}, title = {Heat shock factor 1 over-expression protects against exposure of hydrophobic residues on mutant SOD1 and early mortality in a mouse model of amyotrophic lateral sclerosis.}, journal = {Molecular neurodegeneration}, volume = {8}, number = {}, pages = {43}, pmid = {24256636}, issn = {1750-1326}, support = {NRSA F32 NS063495/NS/NINDS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism ; Animals ; Blotting, Western ; DNA-Binding Proteins/*metabolism ; Disease Models, Animal ; Electrophoresis, Gel, Two-Dimensional ; Fluorescent Antibody Technique ; Heat Shock Transcription Factors ; Humans ; Hydrophobic and Hydrophilic Interactions ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Proteomics/methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spinal Cord/metabolism/pathology ; Superoxide Dismutase/*chemistry/genetics/metabolism ; Superoxide Dismutase-1 ; Transcription Factors/*metabolism ; }, abstract = {BACKGROUND: Mutations in the Cu/Zn superoxide dismutase gene (SOD1) are responsible for 20% of familial forms of amyotrophic lateral sclerosis (ALS), and mutant SOD1 has been shown to have increased surface hydrophobicity in vitro. Mutant SOD1 may adopt a complex array of conformations with varying toxicity in vivo. We have used a novel fluorescence-based proteomic assay using 4,4'-bis-1-anilinonaphthalene-8-sulfonate (bisANS) to assess the surface hydrophobicity, and thereby distinguish between different conformations, of SOD1 and other proteins in situ.<br><br>RESULTS: Covalent bisANS labeling of spinal cord extracts revealed that alterations in surface hydrophobicity of H46R/H48Q mutations in SOD1 provoke formation of high molecular weight SOD1 species with lowered solubility, likely due to increased exposure of hydrophobic surfaces. BisANS was docked on the H46R/H48Q SOD1 structure at the disordered copper binding and electrostatic loops of mutant SOD1, but not non-mutant WT SOD1. 16 non-SOD1 proteins were also identified that exhibited altered surface hydrophobicity in the H46R/H48Q mutant mouse model of ALS, including proteins involved in energy metabolism, cytoskeleton, signaling, and protein quality control. Heat shock proteins (HSPs) were also enriched in the detergent-insoluble fractions with SOD1. Given that chaperones recognize proteins with exposed hydrophobic surfaces as substrates and the importance of protein homeostasis in ALS, we crossed SOD1 H46R/H48Q mutant mice with mice over-expressing the heat shock factor 1 (HSF1) transcription factor. Here we showed that HSF1 over-expression in H46R/H48Q ALS mice enhanced proteostasis as evidenced by increased expression of HSPs in motor neurons and astrocytes and increased solubility of mutant SOD1. HSF1 over-expression significantly reduced body weight loss, delayed ALS disease onset, decreases cases of early disease, and increased survival for the 25th percentile in an H46R/H48Q SOD1 background. HSF1 overexpression did not affect macroautophagy in the ALS background, but was associated with maintenance of carboxyl terminus of Hsp70 interacting protein (CHIP) expression which declined in H46R/H48Q mice.<br><br>CONCLUSION: Our results uncover the potential importance of changes in protein surface hydrophobicity of SOD1 and other non-SOD1 proteins in ALS, and how strategies that activate HSF1 are valid therapies for ALS and other age-associated proteinopathies.}, }
@article {pmid24265138, year = {2014}, author = {Sun, M and Yamashita, T and Shang, J and Liu, N and Deguchi, K and Liu, W and Ikeda, Y and Feng, J and Abe, K}, title = {Acceleration of TDP43 and FUS/TLS protein expressions in the preconditioned hippocampus following repeated transient ischemia.}, journal = {Journal of neuroscience research}, volume = {92}, number = {1}, pages = {54-63}, doi = {10.1002/jnr.23301}, pmid = {24265138}, issn = {1097-4547}, mesh = {Animals ; CA1 Region, Hippocampal/*metabolism/pathology ; DNA-Binding Proteins/*metabolism ; Gerbillinae ; HSP72 Heat-Shock Proteins/metabolism ; Ischemic Attack, Transient/*metabolism/pathology ; *Ischemic Preconditioning ; Male ; Neurons/metabolism/pathology ; RNA-Binding Protein FUS/*metabolism ; Time Factors ; }, abstract = {The 43-kDa transactivation response DNA binding protein (TDP43), fused in sarcoma/translocated in liposarcoma (FUS/TLS), heat shock protein 70 (HSP70), and β-amyloid (Aβ) are induced and involved in cerebral ischemia, amyotrophic lateral sclerosis (ALS), and Alzheimer's disease (AD), but their relationships in ischemic tolerance have never been examined, although they could be involved in endogenous neuroprotection under ischemic preconditioning. In the present study, Mongolian gerbils were subjected to one or three incidents of basically nonlethal 2-min transient common carotid arteries occlusion (tCCAO). Hippocampal CA1 neurons were lost only in the 2-min three times group at 3 and 7 days, which then gradually recovered from 1 to 6 months. Inductions of TDP43 and FUS/TLS were accelerated from 3 months to 7 days or from 7 days to 1 day, respectively, after 2-min three times ischemia compared with once. The cytoplasmic stainings of TDP43 and FUS/TLS showed a further acceleration of the peaks from 1 months to 3 days or from 1 months to 7 days, respectively, after 2-min three times ischemia compared with once. In contrast, HSP70 was induced only at 7 days after 2-min tCCAO for three times, with no expression for Aβ. These data show that ischemic preconditioning offers a way to induce endogenous neuroprotection and neurogenesis in gerbils, with TDP43, FUS/TLS, and HSP70 involved in this function.}, }
@article {pmid24326187, year = {2014}, author = {Sanhueza, M and Zechini, L and Gillespie, T and Pennetta, G}, title = {Gain-of-function mutations in the ALS8 causative gene VAPB have detrimental effects on neurons and muscles.}, journal = {Biology open}, volume = {3}, number = {1}, pages = {59-71}, pmid = {24326187}, issn = {2046-6390}, abstract = {Amyotrophic Lateral Sclerosis (ALS) is a motor neuron degenerative disease characterized by a progressive, and ultimately fatal, muscle paralysis. The human VAMP-Associated Protein B (hVAPB) is the causative gene of ALS type 8. Previous studies have shown that a loss-of-function mechanism is responsible for VAPB-induced ALS. Recently, a novel mutation in hVAPB (V234I) has been identified but its pathogenic potential has not been assessed. We found that neuronal expression of the V234I mutant allele in Drosophila (DVAP-V260I) induces defects in synaptic structure and microtubule architecture that are opposite to those associated with DVAP mutants and transgenic expression of other ALS-linked alleles. Expression of DVAP-V260I also induces aggregate formation, reduced viability, wing postural defects, abnormal locomotion behavior, nuclear abnormalities, neurodegeneration and upregulation of the heat-shock-mediated stress response. Similar, albeit milder, phenotypes are associated with the overexpression of the wild-type protein. These data show that overexpressing the wild-type DVAP is sufficient to induce the disease and that DVAP-V260I is a pathogenic allele with increased wild-type activity. We propose that a combination of gain- and loss-of-function mechanisms is responsible for VAPB-induced ALS.}, }
@article {pmid24563850, year = {2014}, author = {Niforou, K and Cheimonidou, C and Trougakos, IP}, title = {Molecular chaperones and proteostasis regulation during redox imbalance.}, journal = {Redox biology}, volume = {2}, number = {}, pages = {323-332}, pmid = {24563850}, issn = {2213-2317}, mesh = {Animals ; Electron Transport ; Endoplasmic Reticulum/metabolism ; Endoplasmic Reticulum Chaperone BiP ; Free Radicals/metabolism ; Heat-Shock Proteins/physiology ; Homeostasis ; Humans ; Models, Biological ; Molecular Chaperones/*physiology ; Oxidants/metabolism ; Oxidation-Reduction ; Oxidative Stress/*physiology ; Proteasome Endopeptidase Complex/metabolism ; Protein Folding ; Protein Processing, Post-Translational ; Protein Stability ; Proteins/*metabolism ; Proteome ; Reactive Nitrogen Species/metabolism ; Reactive Oxygen Species/metabolism ; }, abstract = {Free radicals originate from both exogenous environmental sources and as by-products of the respiratory chain and cellular oxygen metabolism. Sustained accumulation of free radicals, beyond a physiological level, induces oxidative stress that is harmful for the cellular homeodynamics as it promotes the oxidative damage and stochastic modification of all cellular biomolecules including proteins. In relation to proteome stability and maintenance, the increased concentration of oxidants disrupts the functionality of cellular protein machines resulting eventually in proteotoxic stress and the deregulation of the proteostasis (homeostasis of the proteome) network (PN). PN curates the proteome in the various cellular compartments and the extracellular milieu by modulating protein synthesis and protein machines assembly, protein recycling and stress responses, as well as refolding or degradation of damaged proteins. Molecular chaperones are key players of the PN since they facilitate folding of nascent polypeptides, as well as holding, folding, and/or degradation of unfolded, misfolded, or non-native proteins. Therefore, the expression and the activity of the molecular chaperones are tightly regulated at both the transcriptional and post-translational level at organismal states of increased oxidative and, consequently, proteotoxic stress, including ageing and various age-related diseases (e.g. degenerative diseases and cancer). In the current review we present a synopsis of the various classes of intra- and extracellular chaperones, the effects of oxidants on cellular homeodynamics and diseases and the redox regulation of chaperones.}, }
@article {pmid24595055, year = {2014}, author = {Carlomagno, Y and Zhang, Y and Davis, M and Lin, WL and Cook, C and Dunmore, J and Tay, W and Menkosky, K and Cao, X and Petrucelli, L and Deture, M}, title = {Casein kinase II induced polymerization of soluble TDP-43 into filaments is inhibited by heat shock proteins.}, journal = {PloS one}, volume = {9}, number = {3}, pages = {e90452}, pmid = {24595055}, issn = {1932-6203}, support = {1R21NS079807-01A1/NS/NINDS NIH HHS/United States ; R21 NS079807/NS/NINDS NIH HHS/United States ; R01 NS 063964-01/NS/NINDS NIH HHS/United States ; ES20395-01/ES/NIEHS NIH HHS/United States ; R01 NS063964/NS/NINDS NIH HHS/United States ; R01AG026251/AG/NIA NIH HHS/United States ; R01 ES020395/ES/NIEHS NIH HHS/United States ; R01 AG026251/AG/NIA NIH HHS/United States ; R01 NS077402/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Casein Kinase II/*pharmacology ; DNA-Binding Proteins/*chemistry/isolation &amp; purification/*metabolism ; Frontotemporal Lobar Degeneration/metabolism/pathology ; HSP90 Heat-Shock Proteins/*metabolism ; Humans ; Inclusion Bodies/drug effects/metabolism/ultrastructure ; Mice ; Phosphorylation/drug effects ; Polymerization/*drug effects ; Protein Aggregation, Pathological/metabolism ; Reproducibility of Results ; Solubility ; }, abstract = {BACKGROUND: Trans-activation Response DNA-binding Protein-43 (TDP-43) lesions are observed in Amyotrophic Lateral Sclerosis (ALS), Frontotemporal Lobar Degeneration with ubiquitin inclusions (FTLD-TDP) and 25-50% of Alzheimer's Disease (AD) cases. These abnormal protein inclusions are composed of either amorphous TDP-43 aggregates or highly ordered filaments. The filamentous TDP-43 accumulations typically contain clean 10-12 nm filaments though wider 18-20 nm coated filaments may be observed. The TDP-43 present within these lesions is phosphorylated, truncated and ubiquitinated, and these modifications appear to be abnormal as they are linked to both a cellular heat shock response and microglial activation. The mechanisms associated with this abnormal TDP-43 accumulation are believed to result in a loss of TDP-43 function, perhaps due to the post-translational modifications or resulting from physical sequestration of the TDP-43. The formation of TDP-43 inclusions involves cellular translocation and conversion of TDP-43 into fibrillogenic forms, but the ability of these accumulations to sequester normal TDP-43 and propagate this behavior between neurons pathologically is mostly inferred. The lack of methodology to produce soluble full length TDP-43 and recapitulate this polymerization into filaments as observed in disease has limited our understanding of these pathogenic cascades.<br><br>RESULTS: The protocols described here generate soluble, full-length and untagged TDP-43 allowing for a direct assessment of the impact of various posttranslational modifications on TDP-43 function. We demonstrate that Casein Kinase II (CKII) promotes the polymerization of this soluble TDP-43 into 10 nm diameter filaments that resemble the most common TDP-43 structures observed in disease. Furthermore, these filaments are recognized as abnormal by Heat Shock Proteins (HSPs) which can inhibit TDP-43 polymerization or directly promote TDP-43 filament depolymerization.<br><br>CONCLUSION: These findings demonstrate CKII induces polymerization of soluble TDP-43 into filaments and Hsp90 promotes TDP-43 filament depolymerization. These findings provide rational for potential therapeutic intervention at these points in TDP-43 proteinopathies.}, }
@article {pmid24662024, year = {2014}, author = {Lovett, MC and Coates, JR and Shu, Y and Oglesbee, MJ and Fenner, W and Moore, SA}, title = {Quantitative assessment of hsp70, IL-1β and TNF-α in the spinal cord of dogs with E40K SOD1-associated degenerative myelopathy.}, journal = {Veterinary journal (London, England : 1997)}, volume = {200}, number = {2}, pages = {312-317}, doi = {10.1016/j.tvjl.2014.03.003}, pmid = {24662024}, issn = {1532-2971}, mesh = {Animals ; Biomarkers/cerebrospinal fluid/*metabolism ; CD18 Antigens/genetics/metabolism ; Dog Diseases/metabolism/pathology ; Dogs ; Enzyme-Linked Immunosorbent Assay/veterinary ; *Gene Expression ; HSP70 Heat-Shock Proteins/cerebrospinal fluid/*metabolism ; Immunohistochemistry/veterinary ; Interleukin-1beta/cerebrospinal fluid/*metabolism ; Neurodegenerative Diseases/genetics/metabolism/*veterinary ; Spinal Cord/metabolism/pathology ; Spinal Cord Diseases/genetics/metabolism/*veterinary ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1 ; Tumor Necrosis Factor-alpha/cerebrospinal fluid ; }, abstract = {Inflammation is involved in the pathogenesis of many neurodegenerative diseases. Canine degenerative myelopathy (DM) is a progressive adult-onset neurodegenerative disease commonly associated with an E40K missense mutation in the SOD1 gene. DM has many similarities to some familial forms of human amyotrophic lateral sclerosis (ALS) and may serve as an important disease model for therapy development. Pro-inflammatory mediators such as interleukin (IL)-1β, tumor necrosis factor (TNF)-α and heat shock protein (hsp) 70 play a role in the pathogenesis of ALS. The focus of the current work was to determine whether an inflammatory phenotype is present in canine DM as defined by IL-1β, TNF-α, and hsp70 responses in cerebrospinal fluid (CSF) and spinal cord tissue. Concentrations of hsp70, IL-1β and TNF-α were below the limits of detection by ELISA in the CSF of both normal and DM-affected dogs. Immunohistochemical staining for hsp70 was significantly increased in ependymal cells lining the spinal cord central canal of DM-affected dogs (P = 0.003). This was not associated with increased IL-1β or TNF-α staining, but was associated with increased CD18 staining in the gray matter of DM-affected dogs. These results suggest that hsp70 in spinal cord tissue is a potential inflammatory signature in canine DM.}, }
@article {pmid24685484, year = {2014}, author = {Bhattacharya, A and Wei, R and Hamilton, RT and Chaudhuri, AR}, title = {Neuronal cells but not muscle cells are resistant to oxidative stress mediated protein misfolding and cell death: role of molecular chaperones.}, journal = {Biochemical and biophysical research communications}, volume = {446}, number = {4}, pages = {1250-1254}, doi = {10.1016/j.bbrc.2014.03.097}, pmid = {24685484}, issn = {1090-2104}, support = {K01 AG038555/AG/NIA NIH HHS/United States ; K01AG038555/AG/NIA NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/metabolism ; Animals ; Cell Death ; Cell Line ; Cell Survival ; Cells, Cultured ; Heat-Shock Proteins/analysis/*metabolism ; Mice ; Mice, Inbred C57BL ; Muscle Cells/*cytology/metabolism ; Neurons/*cytology/metabolism ; *Oxidative Stress ; *Protein Folding ; }, abstract = {Our recent study in a mouse model of familial-Amyotrophic Lateral Sclerosis (f-ALS) revealed that muscle proteins are equally sensitive to misfolding as spinal cord proteins despite the presence of low mutant CuZn-superoxide dismutase, which is considered to be the key toxic element for initiation and progression of f-ALS. More importantly, we observed differential level of heat shock proteins (Hsp's) between skeletal muscle and spinal cord tissues prior to the onset and during disease progression; spinal cord maintains significantly higher level of Hsp's compared to skeletal muscle. In this study, we report two important observations; (i) muscle cells (but not neuronal cells) are extremely vulnerable to protein misfolding and cell death during challenge with oxidative stress and (ii) muscle cells fail to mount Hsp's during challenge unlike neuronal cells. These two findings can possibly explain why muscle atrophy precedes the death of motor neurons in f-ALS mice.}, }
@article {pmid25001311, year = {2014}, author = {Trippier, PC and Zhao, KT and Fox, SG and Schiefer, IT and Benmohamed, R and Moran, J and Kirsch, DR and Morimoto, RI and Silverman, RB}, title = {Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis.}, journal = {ACS chemical neuroscience}, volume = {5}, number = {9}, pages = {823-829}, pmid = {25001311}, issn = {1948-7193}, support = {1R43NS057849/NS/NINDS NIH HHS/United States ; }, mesh = {Adaptor Proteins, Signal Transducing ; Amyotrophic Lateral Sclerosis/*metabolism ; Animals ; Anti-Inflammatory Agents, Non-Steroidal/*pharmacology ; Autophagy-Related Proteins ; Biotinylation ; Cell Cycle Proteins/genetics/metabolism ; Cysteine Proteinase Inhibitors/pharmacology ; Disease Models, Animal ; Enzyme Activation/drug effects ; Hot Temperature ; Humans ; Leupeptins/pharmacology ; Luminescent Proteins/genetics/metabolism ; Models, Molecular ; PC12 Cells ; *Proteomics ; Pyrazolones/*chemistry/*pharmacology ; Rats ; Superoxide Dismutase/genetics ; Tandem Mass Spectrometry ; Ubiquitins/genetics/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a progressive and ultimately fatal neurodegenerative disease. Pyrazolone containing small molecules have shown significant disease attenuating efficacy in cellular and murine models of ALS. Pyrazolone based affinity probes were synthesized to identify high affinity binding partners and ascertain a potential biological mode of action. Probes were confirmed to be neuroprotective in PC12-SOD1(G93A) cells. PC12-SOD1(G93A) cell lysates were used for protein pull-down, affinity purification, and subsequent proteomic analysis using LC-MS/MS. Proteomics identified the 26S proteasome regulatory subunit 4 (PSMC1), 26S proteasome regulatory subunit 6B (PSMC4), and T-complex protein 1 (TCP-1) as putative protein targets. Coincubation with appropriate competitors confirmed the authenticity of the proteomics results. Activation of the proteasome by pyrazolones was demonstrated in the absence of exogenous proteasome inhibitor and by restoration of cellular protein degradation of a fluorogenic proteasome substrate in PC12-SOD1(G93A) cells. Importantly, supplementary studies indicated that these molecules do not induce a heat shock response. We propose that pyrazolones represent a rare class of molecules that enhance proteasomal activation in the absence of a heat shock response and may have therapeutic potential in ALS.}, }
@article {pmid25167838, year = {2014}, author = {Watanabe, S and Ageta-Ishihara, N and Nagatsu, S and Takao, K and Komine, O and Endo, F and Miyakawa, T and Misawa, H and Takahashi, R and Kinoshita, M and Yamanaka, K}, title = {SIRT1 overexpression ameliorates a mouse model of SOD1-linked amyotrophic lateral sclerosis via HSF1/HSP70i chaperone system.}, journal = {Molecular brain}, volume = {7}, number = {}, pages = {62}, pmid = {25167838}, issn = {1756-6606}, mesh = {Acetylation ; Amyotrophic Lateral Sclerosis/*metabolism/*pathology ; Animals ; Behavior, Animal ; DNA-Binding Proteins/*metabolism ; Disease Models, Animal ; Gene Dosage ; HSP70 Heat-Shock Proteins/*metabolism ; Heat Shock Transcription Factors ; Humans ; Longevity ; Mice, Inbred C57BL ; Mice, Transgenic ; Mutation/genetics ; Promoter Regions, Genetic/genetics ; Protein Folding ; Sirtuin 1/*metabolism ; Spinal Cord/pathology ; Superoxide Dismutase/*genetics ; Superoxide Dismutase-1 ; Transcription Factors/*metabolism ; Up-Regulation ; }, abstract = {BACKGROUND: Dominant mutations in superoxide dismutase 1 (SOD1) cause degeneration of motor neurons in a subset of inherited amyotrophic lateral sclerosis (ALS). The pathogenetic process mediated by misfolded and/or aggregated mutant SOD1 polypeptides is hypothesized to be suppressed by protein refolding. This genetic study is aimed to test whether mutant SOD1-mediated ALS pathology recapitulated in mice could be alleviated by overexpressing a longevity-related deacetylase SIRT1 whose substrates include a transcription factor heat shock factor 1 (HSF1), the master regulator of the chaperone system.<br><br>RESULTS: We established a line of transgenic mice that chronically overexpress SIRT1 in the brain and spinal cord. While inducible HSP70 (HSP70i) was upregulated in the spinal cord of SIRT1 transgenic mice (PrP-Sirt1), no neurological and behavioral alterations were detected. To test hypothetical benefits of SIRT1 overexpression, we crossbred PrP-Sirt1 mice with two lines of ALS model mice: A high expression line that exhibits a severe phenotype (SOD1G93A-H) or a low expression line with a milder phenotype (SOD1G93A-L). The Sirt1 transgene conferred longer lifespan without altering the time of symptomatic onset in SOD1G93A-L. Biochemical analysis of the spinal cord revealed that SIRT1 induced HSP70i expression through deacetylation of HSF1 and that SOD1G93A-L/PrP-Sirt1 double transgenic mice contained less insoluble SOD1 than SOD1G93A-L mice. Parallel experiments showed that Sirt1 transgene could not rescue a more severe phenotype of SOD1G93A-H transgenic mice partly because their HSP70i level had peaked out.<br><br>CONCLUSIONS: The genetic supplementation of SIRT1 can ameliorate a mutant SOD1-linked ALS mouse model partly through the activation of the HSF1/HSP70i chaperone system. Future studies shall include testing potential benefits of pharmacological enhancement of the deacetylation activity of SIRT1 after the onset of the symptom.}, }
@article {pmid25281879, year = {2015}, author = {Lue, LF and Schmitz, C and Walker, DG}, title = {What happens to microglial TREM2 in Alzheimer's disease: Immunoregulatory turned into immunopathogenic?.}, journal = {Neuroscience}, volume = {302}, number = {}, pages = {138-150}, doi = {10.1016/j.neuroscience.2014.09.050}, pmid = {25281879}, issn = {1873-7544}, support = {P30AG19610/AG/NIA NIH HHS/United States ; }, mesh = {Alzheimer Disease/*genetics/*pathology ; Gene Expression Regulation/physiology ; Humans ; Membrane Glycoproteins/*genetics ; Microglia/metabolism/pathology ; Mutation/*genetics ; Receptors, Immunologic/*genetics ; }, abstract = {Microglia play major roles in initiation, coordination and execution of innate immunity in the brain. In the adult brain, these include maintenance of homeostasis, neuron and tissue repair, and eliminating infectious agents, apoptotic cells, and misfolded proteins. Some of these activities are accompanied by inflammatory reactions; and others are performed with no inflammatory effects. Under normal conditions, triggering receptor expressed on myeloid cells 2 (TREM2) belongs to the second category. It pairs with the adaptor protein DNAX-activating protein of 12kDa (DAP12) to induce phagocytosis of apoptotic neurons without inflammatory responses, and to regulate Toll-like receptor-mediated inflammatory responses, and microglial activation. Although ligands for TREM2 are largely unknown, the mitochondrial heat shock protein 60, expressed on cell surface of apoptotic neurons, is a specific ligand that activates TREM2-mediated phagocytosis by microglia. TREM2 also phagocytoses amyloid beta peptide in cultured cells. Several TREM2 mutations have been identified recently that increase the risk of Alzheimer's disease, Frontotemporal dementia, Parkinson's disease, and amyotrophic lateral sclerosis. Some of these mutations cause impaired proteolysis of full-length TREM2 at the plasma membrane to different degrees. The defects in the intramembrane cleavage result in dysfunction of phagocytosis signaling. The association of TREM2 mutations with neurodegenerative disease also calls for the understanding of the biology and pathological role of non-mutated TREM2 on human brains and microglia. This review provides a summary of current literature in TREM2 and DAP12 from several aspects, and proposes a theory that loss of TREM2 functions might contribute to the immunopathogenic role of microglia in Alzheimer's disease.}, }
@article {pmid25404049, year = {2015}, author = {Patel, P and Julien, JP and Kriz, J}, title = {Early-stage treatment with Withaferin A reduces levels of misfolded superoxide dismutase 1 and extends lifespan in a mouse model of amyotrophic lateral sclerosis.}, journal = {Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics}, volume = {12}, number = {1}, pages = {217-233}, pmid = {25404049}, issn = {1878-7479}, support = {//Canadian Institutes of Health Research/Canada ; }, mesh = {Amyotrophic Lateral Sclerosis/*genetics ; Animals ; Blotting, Western ; Disease Models, Animal ; Fluorescent Antibody Technique ; Immunoprecipitation ; Longevity/*drug effects ; Mice ; Mice, Transgenic ; Motor Neurons/drug effects/pathology ; Neuroprotective Agents/*pharmacology ; Superoxide Dismutase/*genetics ; Superoxide Dismutase-1 ; Withanolides/*pharmacology ; }, abstract = {Approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Recent studies have shown that Withaferin A (WA), an inhibitor of nuclear factor-kappa B activity, was efficient in reducing disease phenotype in a TAR DNA binding protein 43 transgenic mouse model of ALS. These findings led us to test WA in mice from 2 transgenic lines expressing different ALS-linked SOD1 mutations, SOD1(G93A) and SOD1(G37R). Intraperitoneal administration of WA at a dosage of 4 mg/kg of body weight was initiated from postnatal day 40 until end stage in SOD1(G93A) mice, and from 9 months until end stage in SOD1(G37R) mice. The beneficial effects of WA in the SOD1(G93A) mice model were accompanied by an alleviation of neuroinflammation, a decrease in levels of misfolded SOD1 species in the spinal cord, and a reduction in loss of motor neurons resulting in delayed disease progression and mortality. Interestingly, WA treatment triggered robust induction of heat shock protein 25 (a mouse ortholog of heat shock protein 27), which may explain the reduced level of misfolded SOD1 species in the spinal cord of SOD1(G93A) mice and the decrease of neuronal injury responses, as revealed by real-time imaging of biophotonic SOD1(G93A) mice expressing a luciferase transgene under the control of the growth-associated protein 43 promoter. These results suggest that WA may represent a potential lead compound for drug development aiming to treat ALS.}, }
@article {pmid25531407, year = {2014}, author = {Buchan, JR}, title = {mRNP granules. Assembly, function, and connections with disease.}, journal = {RNA biology}, volume = {11}, number = {8}, pages = {1019-1030}, pmid = {25531407}, issn = {1555-8584}, mesh = {Cytoplasmic Granules/genetics/*metabolism ; Gene Expression Regulation ; Humans ; Neurodegenerative Diseases/*genetics/pathology ; Neuronal Plasticity/genetics ; Neurons/metabolism/pathology ; Protein Biosynthesis/genetics ; RNA Transport/*genetics ; RNA, Messenger/*genetics/metabolism ; Ribonucleoproteins/*genetics/metabolism ; Synapses/genetics/metabolism/pathology ; }, abstract = {Messenger ribonucleoprotein (mRNP) granules are dynamic, self-assembling structures that harbor non-translating mRNAs bound by various proteins that regulate mRNA translation, localization, and turnover. Their importance in gene expression regulation is far reaching, ranging from precise spatial-temporal control of mRNAs that drive developmental programs in oocytes and embryos, to similarly exquisite control of mRNAs in neurons that underpin synaptic plasticity, and thus, memory formation. Analysis of mRNP granules in their various contexts has revealed common themes of assembly, disassembly, and modes of mRNA regulation, yet new studies continue to reveal unexpected and important findings, such as links between aberrant mRNP granule assembly and neurodegenerative disease. Continued study of these enigmatic structures thus promises fascinating new insights into cellular function, and may also suggest novel therapeutic strategies in various disease states.}, }
@article {pmid25534414, year = {2015}, author = {Zhao, Z and Sui, Y and Gao, W and Cai, B and Fan, D}, title = {Effects of diet on adenosine monophosphate-activated protein kinase activity and disease progression in an amyotrophic lateral sclerosis model.}, journal = {The Journal of international medical research}, volume = {43}, number = {1}, pages = {67-79}, doi = {10.1177/0300060514554725}, pmid = {25534414}, issn = {1473-2300}, mesh = {Acetyl-CoA Carboxylase/metabolism ; Adenylate Kinase/*metabolism ; Amyotrophic Lateral Sclerosis/*enzymology/*pathology ; Animals ; Body Weight ; Caloric Restriction ; *Diet ; Diet, High-Fat ; Disease Models, Animal ; *Disease Progression ; Feeding Behavior ; Humans ; Male ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/enzymology/pathology ; Nerve Degeneration/pathology ; Phosphorylation ; Superoxide Dismutase/metabolism ; Superoxide Dismutase-1 ; }, abstract = {OBJECTIVES: To study the effects of diet on disease progression and activity levels of adenosine monophosphate-activated protein kinase (AMPK), and its downstream targets, in an amyotrophic lateral sclerosis (ALS) animal model.<br><br>METHODS: AMPK activity was measured in cerebral cortex, spinal cord, cerebellum and hindlimb muscle tissue using immunohistochemistry in transgenic mice overexpressing human superoxide dismutase-1 (SOD1(G93A)) fed a high-fat (HFD), standard ad libitum (AL) or calorie-restricted (CR) diet; AMPK activity was also measured in wild-type (SOD1(WT)) mice. Activity of AMPK and phospho-AMPK, acetyl coenzyme-A carboxylase (ACC), phospho-ACC and heat shock protein-70 (Hsp70) were also measured using Western blot. Food intake and grip strength were recorded; body composition was analysed using dual energy X-ray absorptiometry. Motor neuron survival was observed using Nissl staining.<br><br>RESULTS: AMPK activity increased and Hsp70 expression decreased in AL SOD1(G93A) mice compared with SOD1(WT) mice in spinal cord and hindlimb muscle. Compared with AL SOD1(G93A) mice, CR SOD1(G93A) mice showed increased AMPK activity, downregulated Hsp70 expression, reduced motor neuron survival in spinal cord and hindlimb muscle and reduced lifespan; HFD SOD1(G93A) mice showed opposite effects.<br><br>CONCLUSIONS: In this mouse model, increased AMPK activity seems to play a negative role in motor neuron survival, possibly through a novel mechanism involving Hsp70 downregulation. These changes can be modified by diet. Inhibition of AMPK may provide a therapeutic strategy for ALS.}, }
@article {pmid25557022, year = {2015}, author = {Xu, G and Fromholt, S and Ayers, JI and Brown, H and Siemienski, Z and Crosby, KW and Mayer, CA and Janus, C and Borchelt, DR}, title = {Substantially elevating the levels of αB-crystallin in spinal motor neurons of mutant SOD1 mice does not significantly delay paralysis or attenuate mutant protein aggregation.}, journal = {Journal of neurochemistry}, volume = {133}, number = {3}, pages = {452-464}, pmid = {25557022}, issn = {1471-4159}, support = {P01 NS049134/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Humans ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Mice, Transgenic ; Motor Neurons/*metabolism ; Mutant Proteins/*biosynthesis/genetics ; Paralysis/genetics/*metabolism/prevention &amp; control ; Protein Aggregation, Pathological/genetics/*metabolism/prevention &amp; control ; Spinal Cord/metabolism ; *Superoxide Dismutase/genetics ; Superoxide Dismutase-1 ; alpha-Crystallin B Chain/*biosynthesis/genetics ; }, abstract = {There has been great interest in enhancing endogenous protein maintenance pathways such as the heat-shock chaperone response, as it is postulated that enhancing clearance of misfolded proteins could have beneficial disease modifying effects in amyotrophic lateral sclerosis and other neurodegenerative disorders. In cultured cell models of mutant SOD1 aggregation, co-expression of αB-crystallin (αB-crys) has been shown to inhibit the formation of detergent-insoluble forms of mutant protein. Here, we describe the generation of a new line of transgenic mice that express αB-crys at > 6-fold the normal level in spinal cord, with robust increases in immunoreactivity throughout the spinal cord grey matter and, specifically, in spinal motor neurons. Surprisingly, spinal cords of mice expressing αB-crys alone contained 20% more motor neurons per section than littermate controls. Raising αB-crys by these levels in mice transgenic for either G93A or L126Z mutant SOD1 had no effect on the age at which paralysis developed. In the G93A mice, which showed the most robust degree of motor neuron loss, the number of these cells declined by the same proportion as in mice expressing the mutant SOD1 alone. In paralyzed bigenic mice, the levels of detergent-insoluble, misfolded, mutant SOD1 were similar to those of mice expressing mutant SOD1 alone. These findings indicate that raising the levels of αB-crys in spinal motor neurons by 6-fold does not produce the therapeutic effects predicted by cell culture models of mutant SOD1 aggregation. Enhancing the protein chaperone function may present a therapeutic approach to amyotrophic lateral sclerosis caused by mutations in SOD1, and other neurodegenerative disorders characterized by cytosolic protein aggregation. Previous studies in cell models suggested that the chaperone known as αB-crystallin (αB-crys) can prevent mutant SOD1 aggregation. We report that transgenic expression of αB-crys at > 6-fold the normal level in spinal cords of mice expressing mutant SOD1 produces no therapeutic benefit.}, }
@article {pmid25600110, year = {2015}, author = {Bethea, CL and Reddy, AP}, title = {Ovarian steroids regulate gene expression related to DNA repair and neurodegenerative diseases in serotonin neurons of macaques.}, journal = {Molecular psychiatry}, volume = {20}, number = {12}, pages = {1565-1578}, pmid = {25600110}, issn = {1476-5578}, support = {P51 OD011092/OD/NIH HHS/United States ; R01 MH062677/MH/NIMH NIH HHS/United States ; R24 OD011895/OD/NIH HHS/United States ; U54 HD 18185/HD/NICHD NIH HHS/United States ; MH62677/MH/NIMH NIH HHS/United States ; P30 HD018185/HD/NICHD NIH HHS/United States ; P51 OD 011092/OD/NIH HHS/United States ; U54 HD018185/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; DNA Repair/*genetics ; Estradiol/*physiology ; Female ; *Gene Expression Regulation ; Macaca mulatta ; Neurodegenerative Diseases/*genetics ; Ovariectomy ; Progesterone/*physiology ; Serotonergic Neurons/*metabolism ; }, abstract = {Depression often accompanies the perimenopausal transition and it often precedes overt symptomology in common neurodegenerative diseases (NDDs, such as Alzheimer's, Parkinson's, Huntington, amyotrophic lateral sclerosis). Serotonin dysfunction is frequently found in the different etiologies of depression. We have shown that ovariectomized (Ovx) monkeys treated with estradiol (E) for 28 days supplemented with placebo or progesterone (P) on days 14-28 had reduced DNA fragmentation in serotonin neurons of the dorsal raphe nucleus, and long-term Ovx monkeys had fewer serotonin neurons than intact controls. We questioned the effect of E alone or E+P (estradiol supplemented with progesterone) on gene expression related to DNA repair, protein folding (chaperones), the ubiquitin-proteosome, axon transport and NDD-specific genes in serotonin neurons. Ovx macaques were treated with placebo, E or E+P (n=3 per group) for 1 month. Serotonin neurons were laser captured and subjected to microarray analysis and quantitative real-time PCR (qRT-PCR). Increases were confirmed with qRT-PCR in five genes that code for proteins involved in repair of strand breaks and nucleotide excision. NBN1, PCNA (proliferating nuclear antigen), GADD45A (DNA damage-inducible), RAD23A (DNA damage recognition) and GTF2H5 (gene transcription factor 2H5) significantly increased with E or E+P treatment (all analysis of variance (ANOVA), P<0.01). Chaperone genes HSP70 (heat-shock protein 70), HSP60 and HSP27 significantly increased with E or E+P treatment (all ANOVA, P<0.05). HSP90 showed a similar trend. Ubiquinase coding genes UBEA5, UBE2D3 and UBE3A (Parkin) increased with E or E+P (all ANOVA, P<0.003). Transport-related genes coding kinesin, dynein and dynactin increased with E or E+P treatment (all ANOVA, P<0.03). SCNA (α-synuclein) and ADAM10 (α-secretase) increased (both ANOVA, P<0.02) but PSEN1 (presenilin1) decreased (ANOVA, P<0.02) with treatment. APP decreased 10-fold with E or E+P administration. Newman-Keuls post hoc comparisons indicated variation in the response to E alone versus E+P across the different genes. In summary, E or E+P increased gene expression for DNA repair mechanisms in serotonin neurons, thereby rendering them less vulnerable to stress-induced DNA fragmentation. In addition, E or E+P regulated four genes encoding proteins that are often misfolded or malfunctioning in neuronal populations subserving overt NDD symptomology. The expression and regulation of these genes in serotonergic neurons invites speculation that they may mediate an underlying disease process in NDDs, which in turn may be ameliorated or delayed with timely hormone therapy in women.}, }
@article {pmid25621016, year = {2014}, author = {Zhang, X and Shi, J and Tian, J and Robinson, AC and Davidson, YS and Mann, DM}, title = {Expression of one important chaperone protein, heat shock protein 27, in neurodegenerative diseases.}, journal = {Alzheimer's research &amp; therapy}, volume = {6}, number = {9}, pages = {78}, pmid = {25621016}, issn = {1758-9193}, abstract = {INTRODUCTION: Many neurodegenerative diseases are characterised by accumulations of misfolded proteins that can colocalise with chaperone proteins (for example, heat shock protein 27 (HSP27)), which might act as modulators of protein aggregation.<br><br>METHODS: The role of HSP27 in the pathogenesis of neurodegenerative disorders such as frontotemporal lobar degeneration (FTLD), Alzheimer's disease (AD) and motor neuron disease (MND) was investigated. We used immunohistochemical and Western blot analysis to determine the distribution and amount of this protein in the frontal and temporal cortices of diseased and control subjects.<br><br>RESULTS: HSP27 immunostaining presented as accumulations of granules within neuronal and glial cell perikarya. Patients with AD and FTLD were affected more often, and showed greater immunostaining for HSP27, than patients with MND and controls. In FTLD, there was no association between HSP27 and histological type. The neuropathological changes of FTLD, AD and MND were not immunoreactive to HSP27. Western blot analysis revealed higher HSP27 expression in FTLD than in controls, but without qualitative differences in banding patterns.<br><br>CONCLUSIONS: The pattern of HSP27 immunostaining observed may reflect the extent of ongoing neurodegeneration in affected brain areas and is not specific to FTLD, AD or MND. It may represent an accumulation of misfolded, damaged or unwanted proteins, awaiting or undergoing degradation.}, }
@article {pmid25911031, year = {2015}, author = {Modrzewska, B and Kurnatowski, P}, title = {Adherence of Candida sp. to host tissues and cells as one of its pathogenicity features.}, journal = {Annals of parasitology}, volume = {61}, number = {1}, pages = {3-9}, pmid = {25911031}, issn = {2299-0631}, mesh = {Animals ; Candida/*physiology ; *Cell Adhesion ; Epithelial Cells/*microbiology/physiology ; Fungal Proteins/genetics/*metabolism ; Gene Expression Regulation, Fungal ; Humans ; }, abstract = {The ability of Candida sp. cells to adhere to the mucosal surfaces of various host organs as well as synthetic materials is an important pathogenicity feature of those fungi which contributes to the development of infection. This property varies depending on the species of the fungus and is the greatest for C. albicans. The process of adhesion depends on plenty of factors related to the fungal and host cells as well as environmental conditions. The main adhesins present on the fungal cell wall are: Als, Epa, Hwp1, but also Eap1, Sun41, Csh1 and probably Hyr1; for adhesion significant are also secreted aspartyl proteases Sap. Various researchers specify a range of genes which contribute to adhesion, such as: CZF1, EFG1, TUP1, TPK1, TPK2, HGC1, RAS1, RIM101, VPS11, ECM1, CKA2, BCR1, BUD2, RSR1, IRS4, CHS2, SCS7, UBI4, UME6, TEC1 and GAT2. Influence for adherence have also heat shock proteins Hsp70, Mediator Middle domain subunit Med31 and morphological transition. Among factors affecting adhesion related to host cells it is necessary to mention fibronectins and integrins (receptors for Candida sp. adhesins), type of epithelial cells, their morphology and differentiation phase. To a lesser degree influence on adhesion have non-specific factors and environmental conditions.}, }
@article {pmid26041991, year = {2015}, author = {Chen, D and Wang, Y and Chin, ER}, title = {Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice.}, journal = {Frontiers in cellular neuroscience}, volume = {9}, number = {}, pages = {170}, pmid = {26041991}, issn = {1662-5102}, abstract = {Mutations in Cu/Zn superoxide dismutase (SOD1) are one of the genetic causes of Amyotrophic Lateral Sclerosis (ALS). Although the primary symptom of ALS is muscle weakness, the link between SOD1 mutations, cellular dysfunction and muscle atrophy and weakness is not well understood. The purpose of this study was to characterize cellular markers of ER stress in skeletal muscle across the lifespan of G93A*SOD1 (ALS-Tg) mice. Muscles were obtained from ALS-Tg and age-matched wild type (WT) mice at 70d (pre-symptomatic), 90d and 120-140d (symptomatic) and analyzed for ER stress markers. In white gastrocnemius (WG) muscle, ER stress sensors PERK and IRE1α were upregulated ~2-fold at 70d and remained (PERK) or increased further (IRE1α) at 120-140d. Phospho-eIF2α, a downstream target of PERK and an inhibitor of protein translation, was increased by 70d and increased further to 12.9-fold at 120-140d. IRE1α upregulation leads to increased splicing of X-box binding protein 1 (XBP-1) to the XBP-1s isoform. XBP-1s transcript was increased at 90d and 120-140d indicating activation of IRE1α signaling. The ER chaperone/heat shock protein Grp78/BiP was upregulated 2-fold at 70d and 90d and increased to 6.1-fold by 120-140d. The ER-stress-specific apoptotic signaling protein CHOP was upregulated 2-fold at 70d and 90d and increased to 13.3-fold at 120-140d indicating progressive activation of an apoptotic signal in muscle. There was a greater increase in Grp78/BiP and CHOP in WG vs. the more oxidative red gastrocnemius (RG) ALS-Tg at 120-140d indicating greater ER stress and apoptosis in fast glycolytic muscle. These data show that the ER stress response is activated in skeletal muscle of ALS-Tg mice by an early pre-symptomatic age and increases with disease progression. These data suggest a mechanism by which myocellular ER stress leads to reduced protein translation and contributes to muscle atrophy and weakness in ALS.}, }
@article {pmid26190973, year = {2015}, author = {Irvin, CW and Kim, RB and Mitchell, CS}, title = {Seeking homeostasis: temporal trends in respiration, oxidation, and calcium in SOD1 G93A Amyotrophic Lateral Sclerosis mice.}, journal = {Frontiers in cellular neuroscience}, volume = {9}, number = {}, pages = {248}, pmid = {26190973}, issn = {1662-5102}, support = {K01 NS069616/NS/NINDS NIH HHS/United States ; R01 NS061696/NS/NINDS NIH HHS/United States ; R21 NS081426/NS/NINDS NIH HHS/United States ; }, abstract = {Impairments in mitochondria, oxidative regulation, and calcium homeostasis have been well documented in numerous Amyotrophic Lateral Sclerosis (ALS) experimental models, especially in the superoxide dismutase 1 glycine 93 to alanine (SOD1 G93A) transgenic mouse. However, the timing of these deficiencies has been debatable. In a systematic review of 45 articles, we examine experimental measurements of cellular respiration, mitochondrial mechanisms, oxidative markers, and calcium regulation. We evaluate the quantitative magnitude and statistical temporal trend of these aggregated assessments in high transgene copy SOD1 G93A mice compared to wild type mice. Analysis of overall trends reveals cellular respiration, intracellular adenosine triphosphate, and corresponding mitochondrial elements (Cox, cytochrome c, complex I, enzyme activity) are depressed for the entire lifespan of the SOD1 G93A mouse. Oxidant markers (H2O2, 8OH2'dG, MDA) are initially similar to wild type but are double that of wild type by the time of symptom onset despite early post-natal elevation of protective heat shock proteins. All aspects of calcium regulation show early disturbances, although a notable and likely compensatory convergence to near wild type levels appears to occur between 40 and 80 days (pre-onset), followed by a post-onset elevation in intracellular calcium. The identified temporal trends and compensatory fluctuations provide evidence that the "cause" of ALS may lay within failed homeostatic regulation, itself, rather than any one particular perturbing event or cellular mechanism. We discuss the vulnerabilities of motoneurons to regulatory instability and possible hypotheses regarding failed regulation and its potential treatment in ALS.}, }
@article {pmid26428534, year = {2015}, author = {Finsterer, J and Mishra, A and Wakil, S and Pennuto, M and Soraru, G}, title = {Mitochondrial implications in bulbospinal muscular atrophy (Kennedy disease).}, journal = {Amyotrophic lateral sclerosis &amp; frontotemporal degeneration}, volume = {17}, number = {1-2}, pages = {112-118}, doi = {10.3109/21678421.2015.1089910}, pmid = {26428534}, issn = {2167-9223}, mesh = {Bulbo-Spinal Atrophy, X-Linked/*pathology/*physiopathology ; Evidence-Based Medicine ; Humans ; Mitochondria/*metabolism/*pathology ; Mitochondrial Diseases/etiology/*pathology/*physiopathology ; Mitochondrial Proteins/metabolism ; Models, Biological ; }, abstract = {There is increasing evidence that mitochondrial functions are secondarily disturbed in bulbospinal muscular atrophy (BSMA). This review focuses on the relation between BSMA and the effect of the expanded polyglutamine (poly-Q) androgen receptor (AR) on mitochondrial functions. Mitochondrial functions in bulbospinal muscular atrophy (SBMA) are affected on the molecular, clinical, and therapeutic level. On the molecular level there is down-regulation of various nuclear-DNA-encoded mitochondrial proteins by mutant androgen receptor (mAR), colocalization of the mAR with various mitochondrial proteins, association of mAR aggregates with mitochondria resulting in abnormal distribution of mitochondria, mtDNA depletion or multiple mtDNA deletions, mitochondrial membrane depolarization, increase in reactive oxidative species, and activation of the mitochondrial caspase pathway. On the clinical level various mitochondrial disorders mimic SBMA, and on the therapeutic level pioglitazone expresses PPAR-γ, cyclosporine-A restores mitochondrial membrane potentials, coenzyme-Q and idebenone reduce oxidative stress, and geldanamycin up-regulates protective mitochondrial heat shock proteins. In conclusion, in BSMA mitochondrial dysfunction results from various interactions of elongated poly-Q AR with mitochondria, mitochondrial proteins, nuclear or mitochondrial DNA, causing oxidative stress, decreased mitochondrial membrane potential, or activation of the mitochondrial caspase pathway. Additionally, mitochondrial disease may mimic BSMA and therapeutic approaches may depend on modifications of mitochondrial pathways.}, }
@article {pmid26713267, year = {2015}, author = {Siklos, M and BenAissa, M and Thatcher, GR}, title = {Cysteine proteases as therapeutic targets: does selectivity matter? A systematic review of calpain and cathepsin inhibitors.}, journal = {Acta pharmaceutica Sinica. B}, volume = {5}, number = {6}, pages = {506-519}, pmid = {26713267}, issn = {2211-3835}, abstract = {Cysteine proteases continue to provide validated targets for treatment of human diseases. In neurodegenerative disorders, multiple cysteine proteases provide targets for enzyme inhibitors, notably caspases, calpains, and cathepsins. The reactive, active-site cysteine provides specificity for many inhibitor designs over other families of proteases, such as aspartate and serine; however, a) inhibitor strategies often use covalent enzyme modification, and b) obtaining selectivity within families of cysteine proteases and their isozymes is problematic. This review provides a general update on strategies for cysteine protease inhibitor design and a focus on cathepsin B and calpain 1 as drug targets for neurodegenerative disorders; the latter focus providing an interesting query for the contemporary assumptions that irreversible, covalent protein modification and low selectivity are anathema to therapeutic safety and efficacy.}, }
@article {pmid26872075, year = {2016}, author = {Yerbury, JJ and Ooi, L and Dillin, A and Saunders, DN and Hatters, DM and Beart, PM and Cashman, NR and Wilson, MR and Ecroyd, H}, title = {Walking the tightrope: proteostasis and neurodegenerative disease.}, journal = {Journal of neurochemistry}, volume = {137}, number = {4}, pages = {489-505}, doi = {10.1111/jnc.13575}, pmid = {26872075}, issn = {1471-4159}, mesh = {Animals ; Humans ; Neurodegenerative Diseases/*metabolism/pathology ; Protein Folding ; Protein Interaction Maps/physiology ; *Proteolysis ; Proteostasis Deficiencies/*metabolism/pathology ; Ubiquitin/metabolism ; }, abstract = {A characteristic of many neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS), is the aggregation of specific proteins into protein inclusions and/or plaques in degenerating brains. While much of the aggregated protein consists of disease specific proteins, such as amyloid-β, α-synuclein, or superoxide dismutase1 (SOD1), many other proteins are known to aggregate in these disorders. Although the role of protein aggregates in the pathogenesis of neurodegenerative diseases remains unknown, the ubiquitous association of misfolded and aggregated proteins indicates that significant dysfunction in protein homeostasis (proteostasis) occurs in these diseases. Proteostasis is the concept that the integrity of the proteome is in fine balance and requires proteins in a specific conformation, concentration, and location to be functional. In this review, we discuss the role of specific mechanisms, both inside and outside cells, which maintain proteostasis, including molecular chaperones, protein degradation pathways, and the active formation of inclusions, in neurodegenerative diseases associated with protein aggregation. A characteristic of many neurodegenerative diseases is the aggregation of specific proteins, which alone provides strong evidence that protein homeostasis is disrupted in these disease states. Proteostasis is the maintenance of the proteome in the correct conformation, concentration, and location by functional pathways such as molecular chaperones and protein degradation machinery. Here, we discuss the potential roles of quality control pathways, both inside and outside cells, in the loss of proteostasis during aging and disease.}, }
@article {pmid26878578, year = {2015}, author = {Nefedova, VV and Muranova, LK and Sudnitsyna, MV and Ryzhavskaya, AS and Gusev, NB}, title = {Small Heat Shock Proteins and Distal Hereditary Neuropathies.}, journal = {Biochemistry. Biokhimiia}, volume = {80}, number = {13}, pages = {1734-1747}, doi = {10.1134/S000629791513009X}, pmid = {26878578}, issn = {1608-3040}, mesh = {Adolescent ; Adult ; Aged ; Charcot-Marie-Tooth Disease/genetics/*metabolism ; Child ; HSP27 Heat-Shock Proteins/*genetics/metabolism ; Heat-Shock Proteins ; Heat-Shock Proteins, Small/genetics ; Humans ; Middle Aged ; Molecular Chaperones ; *Mutation ; Peripheral Nervous System Diseases/genetics/metabolism ; Protein Interaction Domains and Motifs ; Protein Multimerization ; Protein Stability ; Young Adult ; }, abstract = {Classification of small heat shock proteins (sHsp) is presented and processes regulated by sHsp are described. Symptoms of hereditary distal neuropathy are described and the genes whose mutations are associated with development of this congenital disease are listed. The literature data and our own results concerning physicochemical properties of HspB1 mutants associated with Charcot-Marie-Tooth disease are analyzed. Mutations of HspB1, associated with hereditary motor neuron disease, can be accompanied by change of the size of HspB1 oligomers, by decreased stability under unfavorable conditions, by changes in the interaction with protein partners, and as a rule by decrease of chaperone-like activity. The largest part of these mutations is accompanied by change of oligomer stability (that can be either increased or decreased) or by change of intermonomer interaction inside an oligomer. Data on point mutation of HspB3 associated with axonal neuropathy are presented. Data concerning point mutations of Lys141 of HspB8 and those associated with hereditary neuropathy and different forms of Charcot-Marie-Tooth disease are analyzed. It is supposed that point mutations of sHsp associated with distal neuropathies lead either to loss of function (for instance, decrease of chaperone-like activity) or to gain of harmful functions (for instance, increase of interaction with certain protein partners).}, }
@article {pmid26936937, year = {2016}, author = {Chen, HJ and Mitchell, JC and Novoselov, S and Miller, J and Nishimura, AL and Scotter, EL and Vance, CA and Cheetham, ME and Shaw, CE}, title = {The heat shock response plays an important role in TDP-43 clearance: evidence for dysfunction in amyotrophic lateral sclerosis.}, journal = {Brain : a journal of neurology}, volume = {139}, number = {Pt 5}, pages = {1417-1432}, pmid = {26936937}, issn = {1460-2156}, support = {/WT_/Wellcome Trust/United Kingdom ; SHAW/APR15/970-797/MNDA_/Motor Neurone Disease Association/United Kingdom ; 092621/WT_/Wellcome Trust/United Kingdom ; 089701/Z/09/2/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/metabolism/*physiopathology ; Animals ; Cell Survival/genetics ; DNA-Binding Proteins/biosynthesis/*metabolism/*physiology ; HSP40 Heat-Shock Proteins/metabolism/physiology ; Heat Shock Transcription Factors ; Heat-Shock Proteins/*metabolism ; Humans ; Mice ; Mice, Transgenic ; Neurons/metabolism ; Phosphorylation ; Transcription Factors/biosynthesis/*physiology ; Ubiquitin/metabolism ; }, abstract = {Detergent-resistant, ubiquitinated and hyperphosphorylated Tar DNA binding protein 43 (TDP-43, encoded by TARDBP) neuronal cytoplasmic inclusions are the pathological hallmark in ∼95% of amyotrophic lateral sclerosis and ∼60% of frontotemporal lobar degeneration cases. We sought to explore the role for the heat shock response in the clearance of insoluble TDP-43 in a cellular model of disease and to validate our findings in transgenic mice and human amyotrophic lateral sclerosis tissues. The heat shock response is a stress-responsive protective mechanism regulated by the transcription factor heat shock factor 1 (HSF1), which increases the expression of chaperones that refold damaged misfolded proteins or facilitate their degradation. Here we show that manipulation of the heat shock response by expression of dominant active HSF1 results in a dramatic reduction of insoluble and hyperphosphorylated TDP-43 that enhances cell survival, whereas expression of dominant negative HSF1 leads to enhanced TDP-43 aggregation and hyperphosphorylation. To determine which chaperones were mediating TDP-43 clearance we over-expressed a range of heat shock proteins (HSPs) and identified DNAJB2a (encoded by DNAJB2, and also known as HSJ1a) as a potent anti-aggregation chaperone for TDP-43. DNAJB2a has a J domain, allowing it to interact with HSP70, and ubiquitin interacting motifs, which enable it to engage the degradation of its client proteins. Using functionally deleted DNAJB2a constructs we demonstrated that TDP-43 clearance was J domain-dependent and was not affected by ubiquitin interacting motif deletion or proteasome inhibition. This indicates that TDP-43 is maintained in a soluble state by DNAJB2a, leaving the total levels of TDP-43 unchanged. Additionally, we have demonstrated that the levels of HSF1 and heat shock proteins are significantly reduced in affected neuronal tissues from a TDP-43 transgenic mouse model of amyotrophic lateral sclerosis and patients with sporadic amyotrophic lateral sclerosis. This implies that the HSF1-mediated DNAJB2a/HSP70 heat shock response pathway is compromised in amyotrophic lateral sclerosis. Defective refolding of TDP-43 is predicted to aggravate the TDP-43 proteinopathy. The finding that the pathological accumulation of insoluble TDP-43 can be reduced by the activation of HSF1/HSP pathways presents an exciting opportunity for the development of novel therapeutics.}, }
@article {pmid26961006, year = {2016}, author = {Crippa, V and D'Agostino, VG and Cristofani, R and Rusmini, P and Cicardi, ME and Messi, E and Loffredo, R and Pancher, M and Piccolella, M and Galbiati, M and Meroni, M and Cereda, C and Carra, S and Provenzani, A and Poletti, A}, title = {Transcriptional induction of the heat shock protein B8 mediates the clearance of misfolded proteins responsible for motor neuron diseases.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {22827}, pmid = {26961006}, issn = {2045-2322}, mesh = {Amyotrophic Lateral Sclerosis/metabolism ; Animals ; Autophagy ; Cell Line ; Colchicine/*pharmacology ; DNA-Binding Proteins/biosynthesis ; Doxorubicin/*pharmacology ; Frontotemporal Dementia/metabolism ; Heat-Shock Proteins/*biosynthesis ; Heat-Shock Response/drug effects ; High-Throughput Screening Assays ; Humans ; Mice ; Molecular Chaperones ; Motor Neurons/cytology/*drug effects/metabolism ; Peptide Fragments/biosynthesis ; Protein Folding ; Protein Serine-Threonine Kinases/*biosynthesis ; Transcription, Genetic ; }, abstract = {Neurodegenerative diseases (NDs) are often associated with the presence of misfolded protein inclusions. The chaperone HSPB8 is upregulated in mice, the human brain and muscle structures affected during NDs progression. HSPB8 exerts a potent pro-degradative activity on several misfolded proteins responsible for familial NDs forms. Here, we demonstrated that HSPB8 also counteracts accumulation of aberrantly localized misfolded forms of TDP-43 and its 25 KDa fragment involved in most sporadic cases of Amyotrophic Lateral Sclerosis (sALS) and of Fronto Lateral Temporal Dementia (FLTD). HSPB8 acts with BAG3 and the HSP70/HSC70-CHIP complex enhancing the autophagic removal of misfolded proteins. We performed a high-through put screening (HTS) to find small molecules capable of inducing HSPB8 in neurons for therapeutic purposes. We identified two compounds, colchicine and doxorubicin, that robustly up-regulated HSPB8 expression. Both colchicine and doxorubicin increased the expression of the master regulator of autophagy TFEB, the autophagy linker p62/SQSTM1 and the autophagosome component LC3. In line, both drugs counteracted the accumulation of TDP-43 and TDP-25 misfolded species responsible for motoneuronal death in sALS. Thus, analogs of colchicine and doxorubicin able to induce HSPB8 and with better safety and tolerability may result beneficial in NDs models.}, }
@article {pmid26994698, year = {2016}, author = {Lin, PY and Folorunso, O and Taglialatela, G and Pierce, A}, title = {Overexpression of heat shock factor 1 maintains TAR DNA binding protein 43 solubility via induction of inducible heat shock protein 70 in cultured cells.}, journal = {Journal of neuroscience research}, volume = {94}, number = {7}, pages = {671-682}, doi = {10.1002/jnr.23725}, pmid = {26994698}, issn = {1097-4547}, mesh = {Alzheimer Disease/genetics/metabolism ; Amyotrophic Lateral Sclerosis/genetics/metabolism ; Animals ; Autophagy/genetics ; Cell Line, Tumor ; Cells, Cultured ; DNA-Binding Proteins/*chemistry/toxicity ; HSP70 Heat-Shock Proteins/*biosynthesis ; Heat Shock Transcription Factors/biosynthesis/*genetics ; Humans ; Mice ; Mice, Knockout ; Primary Cell Culture ; Proteasome Endopeptidase Complex/drug effects ; Solubility ; Ubiquitin/metabolism ; }, abstract = {TAR DNA binding protein 43 (TDP-43) is a nuclear protein that has been shown to have altered homeostasis in the form of neuronal nuclear and cytoplasmic aggregates in some familial and almost all cases of sporadic amyotrophic lateral sclerosis as well as 51% of frontotemporal lobar degeneration and 57% of Alzheimer's disease cases. Heat shock proteins (HSPs), such as HSP70, recognize misfolded or aggregated proteins and refold, disaggregate, or turn them over and are upregulated by the master transcription factor heat shock factor 1 (HSF1). Here, we explore the effect of HSF1 overexpression on proteotoxic stress-related alterations in TDP-43 solubility, proteolytic processing, and cytotoxicity. HSF1 overexpression reduced TDP-43-positive puncta concomitantly with upregulating HSP70 and HSP90 protein levels. HSF1 overexpression or pharmacological activation sustained TDP-43 solubility and significantly reduced truncation of TDP-43 in response to inhibition of the proteasome with Z-Leu-Leu-Leu-al, and this was reversed by HSF1 inhibition. HSF1 activation conferred protection against toxicity associated with TDP-43 C-terminal fragments without globally increasing the activity of the ubiquitin proteasome system (UPS) while concomitantly reducing the induction of autophagy, suggesting that HSF1 protection is an early event. In support of this, inhibition of HSP70 ATPase activity further reduced TDP-43 solubility. HSF1 knockout significantly increased TDP-43 insolubility and accelerated TDP-43 fragmentation in response to proteotoxic stress. Overall, this study shows that HSF1 overexpression protects against TDP-43 pathology by upregulation of chaperones, especially HSP70, rather than enhancing autophagy or the UPS during times of proteotoxic stress. © 2016 Wiley Periodicals, Inc.}, }
@article {pmid27064076, year = {2016}, author = {Parakh, S and Atkin, JD}, title = {Protein folding alterations in amyotrophic lateral sclerosis.}, journal = {Brain research}, volume = {1648}, number = {Pt B}, pages = {633-649}, doi = {10.1016/j.brainres.2016.04.010}, pmid = {27064076}, issn = {1872-6240}, mesh = {Amyotrophic Lateral Sclerosis/genetics/*metabolism/*physiopathology ; Animals ; C9orf72 Protein/genetics ; DNA-Binding Proteins/genetics ; Humans ; Molecular Chaperones/genetics ; Mutation/genetics ; *Protein Folding ; Proteostasis Deficiencies/genetics ; RNA-Binding Protein FUS/genetics ; Superoxide Dismutase-1/genetics ; }, abstract = {Protein misfolding leads to the formation of aggregated proteins and protein inclusions, which are associated with synaptic loss and neuronal death in neurodegenerative diseases. Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that targets motor neurons in the brain, brainstem and spinal cord. Several proteins misfold and are associated either genetically or pathologically in ALS, including superoxide dismutase 1 (SOD1), Tar DNA binding protein-43 (TDP-43), Ubiquilin-2, p62, VCP, and dipeptide repeat proteins produced by unconventional repeat associated non-ATG translation of the repeat expansion in C9ORF72. Chaperone proteins, including heat shock proteins (Hsp׳s) and the protein disulphide isomerase (PDI) family, assist in protein folding and therefore can prevent protein misfolding, and have been implicated as being protective in ALS. In this review we provide an overview of the current literature regarding the molecular mechanisms of protein misfolding and aggregation in ALS, and the role of chaperones as potential targets for therapeutic intervention. This article is part of a Special Issue entitled SI:ER stress.}, }
@article {pmid27106072, year = {2016}, author = {Kampinga, HH and Bergink, S}, title = {Heat shock proteins as potential targets for protective strategies in neurodegeneration.}, journal = {The Lancet. Neurology}, volume = {15}, number = {7}, pages = {748-759}, doi = {10.1016/S1474-4422(16)00099-5}, pmid = {27106072}, issn = {1474-4465}, mesh = {Animals ; Heat-Shock Proteins/drug effects/*metabolism ; Humans ; Neurodegenerative Diseases/drug therapy/*metabolism ; }, abstract = {Protein aggregates are hallmarks of nearly all age-related neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and several polyglutamine diseases such as Huntington's disease and different forms of spinocerebellar ataxias (SCA; SCA1-3, SCA6, and SCA7). The collapse of cellular protein homoeostasis can be both a cause and a consequence of this protein aggregation. Boosting components of the cellular protein quality control system has been widely investigated as a strategy to counteract protein aggregates or their toxic consequences. Heat shock proteins (HSPs) play a central part in regulating protein quality control and contribute to protein aggregation and disaggregation. Therefore, HSPs are viable targets for the development of drugs aimed at reducing pathogenic protein aggregates that are thought to contribute to the development of so many neurodegenerative disorders.}, }
@article {pmid27112486, year = {2016}, author = {Miyazaki, D and Nakamura, A and Hineno, A and Kobayashi, C and Kinoshita, T and Yoshida, K and Ikeda, S}, title = {Elevation of serum heat-shock protein levels in amyotrophic lateral sclerosis.}, journal = {Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology}, volume = {37}, number = {8}, pages = {1277-1281}, pmid = {27112486}, issn = {1590-3478}, mesh = {Aged ; Amyotrophic Lateral Sclerosis/*blood ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; HSP90 Heat-Shock Proteins ; Heat-Shock Proteins/*blood ; Humans ; Male ; Middle Aged ; }, abstract = {Heat-shock proteins (HSPs) have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). In this study, we aimed to examine whether the serum levels of HSPs (HSP27, HSP70, and HSP90) are altered in patients with ALS. We included 58 patients diagnosed with ALS and 85 control individuals. Serum HSP levels of patients and controls were determined using enzyme-linked immunosorbent assay. The serum levels of HSP70 and HSP90 were significantly higher in patients than in controls. In contrast, serum levels of HSP27 did not differ significantly between the patient and control groups. Moreover, serum levels of HSP70 and HSP90 in patients remained high throughout the duration of the disease. Taken together, our findings suggest that HSPs might have a role in ALS progression throughout the course of the disease. Further studies are needed to clarify the role of HSPs in the pathogenesis of ALS.}, }
@article {pmid27496206, year = {2016}, author = {Healy, EF and Cervantes, L}, title = {An in silico study of the effect of SOD1 electrostatic loop dynamics on amyloid‑like filament formation.}, journal = {European biophysics journal : EBJ}, volume = {45}, number = {8}, pages = {853-859}, pmid = {27496206}, issn = {1432-1017}, support = {K12 GM102745/GM/NIGMS NIH HHS/United States ; }, mesh = {Amyloid beta-Peptides/*chemistry ; *Computer Simulation ; Humans ; Hydrogen Bonding ; Molecular Dynamics Simulation ; Mutation ; Protein Aggregates ; Protein Multimerization ; Protein Structure, Secondary ; *Static Electricity ; Superoxide Dismutase-1/*chemistry/genetics/*metabolism ; alpha-Crystallins/chemistry/metabolism ; }, abstract = {Superoxide dismutase [Cu-Zn], or SOD1, is a homo-dimeric protein that functions as an antioxidant by scavenging for superoxides. A wide range of SOD1 variants are linked to inherited, or familial, amyotrophic lateral sclerosis, a progressive and fatal neurodegenerative disease. Aberrant SOD1 oligomerization has been strongly implicated in disease causation, even for sporadic ALS, or SALS, which accounts for ~90 % of ALS cases. Small heat shock proteins (sHSP) have been shown to protect against amyloid fibril formation in vitro, and the sHSP αB-crystallin suppresses in vitro aggregation of SOD1. We are seeking to elucidate the structural features of both SOD1 amyloid formation and αB-crystallin amyloid suppression. Specifically, we have used a flexible docking protocol to refine our model of a SOD1 non-obligate tetramer, postulated to function as a transient desolvating complex. Homology modeling and molecular dynamics (MD) are used to supply the missing structural elements of a previously characterized SOD1 amyloid filament, thereby providing a structural analysis for the observed gain of interaction. This completed filament is then further modified using MD to provide a structural model for protofibril capping of SOD1 filaments by αB-crystallin.}, }
@article {pmid27671821, year = {2016}, author = {Venkatesha, SH and Moudgil, KD}, title = {Celastrol and Its Role in Controlling Chronic Diseases.}, journal = {Advances in experimental medicine and biology}, volume = {928}, number = {}, pages = {267-289}, pmid = {27671821}, issn = {0065-2598}, support = {R01 AT004321/AT/NCCIH NIH HHS/United States ; }, mesh = {Angiogenesis Inhibitors/pharmacology ; Animals ; Antineoplastic Agents, Phytogenic/pharmacology ; Autoimmune Diseases/drug therapy ; Chronic Disease ; Humans ; Inflammation/drug therapy ; Pentacyclic Triterpenes ; Signal Transduction/drug effects ; Triterpenes/pharmacology/*therapeutic use ; }, abstract = {Celastrol, a triterpenoid derived from traditional Chinese medicinal plants, has anti-inflammatory, antioxidant, and anticancer activities. Celastrol has shown preventive/therapeutic effects in experimental models of several chronic diseases. These include, chronic inflammatory and autoimmune diseases (e.g., rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, and psoriasis), neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's disease, and Amyotrophic lateral sclerosis), atherosclerosis, obesity, Type 2 diabetes, and cancer. Celastrol modulates intricate cellular pathways and networks associated with disease pathology, and it interrupts or redirects the aberrant cellular and molecular events so as to limit disease progression and facilitate recovery, where feasible. The major cell signaling pathways modulated by celastrol include the NF-kB pathway, MAPK pathway, JAK/STAT pathway, PI3K/Akt/mTOR pathway, and antioxidant defense mechanisms. Furthermore, celastrol modulates cell proliferation, apoptosis, proteasome activity, heat-shock protein response, innate and adaptive immune responses, angiogenesis, and bone remodeling. Current understanding of the mechanisms of action of celastrol and information about its disease-modulating activities in experimental models have set the stage for testing celastrol in clinical studies as a therapeutic agent for several chronic human diseases.}, }
@article {pmid27748814, year = {2016}, author = {Shen, H and Kim, K and Oh, Y and Yoon, KS and Baik, HH and Kim, SS and Ha, J and Kang, I and Choe, W}, title = {Neurotoxin β‑N‑methylamino‑L‑alanine induces endoplasmic reticulum stress‑mediated neuronal apoptosis.}, journal = {Molecular medicine reports}, volume = {14}, number = {5}, pages = {4873-4880}, doi = {10.3892/mmr.2016.5802}, pmid = {27748814}, issn = {1791-3004}, mesh = {Amino Acids, Diamino/*pharmacology ; Animals ; Apoptosis/drug effects/genetics ; Cell Line ; Cyanobacteria Toxins ; Endoplasmic Reticulum Stress/*drug effects/genetics ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Humans ; MAP Kinase Signaling System/drug effects ; Mice ; Neurons/*drug effects/*metabolism ; Neurotoxins/*pharmacology ; Signal Transduction/drug effects ; Transcription Factor CHOP/genetics/metabolism ; Unfolded Protein Response/drug effects ; }, abstract = {β-N-methylamino-L-alanine (BMAA) is a neurotoxin that is closely associated with the incidence of amyotrophic lateral sclerosis, Parkinson's disease and Alzheimer's disease. In cultured neuronal cells, BMAA notably induces the upregulation of endoplasmic reticulum (ER) chaperons and activates the unfolded protein response (UPR) receptor pathways of protein kinase RNA‑like endoplasmic reticulum kinase, inositol‑requiring kinase 1 and transcription factor 6. The ER stress‑specific protein CCAAT/‑enhancer‑binding protein homologous protein (CHOP) affords pro‑apoptotic responses that cause mitochondrial damage and caspase activation. BMAA also induces the activation of mitogen‑activated protein kinase member c‑JUN N‑terminal kinase, p38 and extracellular signal‑regulated kinase, which have been suggested to be involved in the signaling pathway of UPR‑mediated apoptosis. Inhibition of ER stress using ER stress antagonist, salubrinal, attenuated the expression of CHOP and alleviated neuronal death. Overexpression of heat shock protein 70 suppressed the activation of UPR receptors and UPR‑evoked apoptotic signaling. The present findings demonstrated that ER stress induced by BMAA is the important mediator of neuronal injury and apoptotic death, and suggests development in novel therapeutic strategies for treatment.}, }
@article {pmid27804858, year = {2017}, author = {van Noort, JM and Bugiani, M and Amor, S}, title = {Heat Shock Proteins: Old and Novel Roles in Neurodegenerative Diseases in the Central Nervous System.}, journal = {CNS &amp; neurological disorders drug targets}, volume = {16}, number = {3}, pages = {244-256}, doi = {10.2174/1871527315666161031125317}, pmid = {27804858}, issn = {1996-3181}, mesh = {Animals ; Central Nervous System/*metabolism ; Heat-Shock Proteins/*metabolism ; Humans ; Neurodegenerative Diseases/*metabolism/*pathology ; }, abstract = {Heat shock proteins (HSPs) are families of molecular chaperones that play important homeostatic functions in the central nervous system (CNS) by preventing protein misfolding, promoting degradation of improperly folded proteins, and protecting against apoptosis and inflammatory damage especially during hyperthermia, hypoxia, or oxidative stress. Under stress conditions, HSPs are upregulated to protect cells from damage that accumulates during ageing as well as pathological conditions. An important, yet frequently overlooked function of some HSPs is their ability to function as extracellular messengers (also termed chaperokines) that modulate immune responses within the CNS. Given the strong association between protein aggregation, innate immune cell activation and neurodegeneration, the expression and roles of HSPs in the CNS is attracting attention in many neurodegenerative disorders including inflammatory diseases such as multiple sclerosis, protein folding diseases such as Alzheimer's disease and amyotrophic lateral sclerosis, and genetic white matter diseases. This is especially so since several studies show that HSPs act therapeutically by modulating innate immune activation and may thus serve as neuroprotective agents. Here we review the evidence linking HSPs with neurodegenerative disorders in humans and the experimental animal models of these disorders. We discuss the mechanisms by which HSPs protect cells, and how the knowledge of their endogenous functions can be exploited to treat disorders of the CNS.}, }
@article {pmid28077174, year = {2017}, author = {Geuens, T and De Winter, V and Rajan, N and Achsel, T and Mateiu, L and Almeida-Souza, L and Asselbergh, B and Bouhy, D and Auer-Grumbach, M and Bagni, C and Timmerman, V}, title = {Mutant HSPB1 causes loss of translational repression by binding to PCBP1, an RNA binding protein with a possible role in neurodegenerative disease.}, journal = {Acta neuropathologica communications}, volume = {5}, number = {1}, pages = {5}, pmid = {28077174}, issn = {2051-5960}, support = {P 27634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Brain/metabolism ; Carrier Proteins/genetics/*metabolism ; Charcot-Marie-Tooth Disease/genetics/metabolism ; Consensus Sequence ; DNA-Binding Proteins ; Fibroblasts/metabolism ; Gene Expression Regulation/physiology ; HEK293 Cells ; HSP27 Heat-Shock Proteins/genetics/*metabolism ; HeLa Cells ; Heat-Shock Proteins/genetics/*metabolism ; Heterogeneous-Nuclear Ribonucleoproteins/genetics/*metabolism ; Humans ; Mice ; Molecular Chaperones ; Mutant Proteins/genetics/metabolism ; Mutation ; Neoplasm Proteins/genetics/*metabolism ; Protein Binding ; Protein Biosynthesis/physiology ; RNA, Messenger/genetics/metabolism ; RNA-Binding Proteins ; Untranslated Regions ; }, abstract = {The small heat shock protein HSPB1 (Hsp27) is an ubiquitously expressed molecular chaperone able to regulate various cellular functions like actin dynamics, oxidative stress regulation and anti-apoptosis. So far disease causing mutations in HSPB1 have been associated with neurodegenerative diseases such as distal hereditary motor neuropathy, Charcot-Marie-Tooth disease and amyotrophic lateral sclerosis. Most mutations in HSPB1 target its highly conserved α-crystallin domain, while other mutations affect the C- or N-terminal regions or its promotor. Mutations inside the α-crystallin domain have been shown to enhance the chaperone activity of HSPB1 and increase the binding to client proteins. However, the HSPB1-P182L mutation, located outside and downstream of the α-crystallin domain, behaves differently. This specific HSPB1 mutation results in a severe neuropathy phenotype affecting exclusively the motor neurons of the peripheral nervous system. We identified that the HSPB1-P182L mutant protein has a specifically increased interaction with the RNA binding protein poly(C)binding protein 1 (PCBP1) and results in a reduction of its translational repressive activity. RNA immunoprecipitation followed by RNA sequencing on mouse brain lead to the identification of PCBP1 mRNA targets. These targets contain larger 3'- and 5'-UTRs than average and are enriched in an RNA motif consisting of the CTCCTCCTCCTCC consensus sequence. Interestingly, next to the clear presence of neuronal transcripts among the identified PCBP1 targets we identified known genes associated with hereditary peripheral neuropathies and hereditary spastic paraplegias. We therefore conclude that HSPB1 can mediate translational repression through interaction with an RNA binding protein further supporting its role in neurodegenerative disease.}, }
@article {pmid28167528, year = {2017}, author = {Li, W and Reeb, AN and Lin, B and Subramanian, P and Fey, EE and Knoverek, CR and French, RL and Bigio, EH and Ayala, YM}, title = {Heat Shock-induced Phosphorylation of TAR DNA-binding Protein 43 (TDP-43) by MAPK/ERK Kinase Regulates TDP-43 Function.}, journal = {The Journal of biological chemistry}, volume = {292}, number = {12}, pages = {5089-5100}, pmid = {28167528}, issn = {1083-351X}, support = {K01 NS082391/NS/NINDS NIH HHS/United States ; P30 AG013854/AG/NIA NIH HHS/United States ; }, mesh = {DNA-Binding Proteins/*metabolism ; HEK293 Cells/chemistry ; HeLa Cells ; Heat-Shock Response ; Humans ; MAP Kinase Kinase Kinases/metabolism ; *MAP Kinase Signaling System ; Models, Molecular ; Phosphorylation ; Protein Aggregates ; Ribonucleoside Diphosphate Reductase ; Tumor Suppressor Proteins/metabolism ; }, abstract = {TAR DNA-binding protein (TDP-43) is a highly conserved and essential DNA- and RNA-binding protein that controls gene expression through RNA processing, in particular, regulation of splicing. Intracellular aggregation of TDP-43 is a hallmark of amyotrophic lateral sclerosis and ubiquitin-positive frontotemporal lobar degeneration. This TDP-43 pathology is also present in other types of neurodegeneration including Alzheimer's disease. We report here that TDP-43 is a substrate of MEK, a central kinase in the MAPK/ERK signaling pathway. TDP-43 dual phosphorylation by MEK, at threonine 153 and tyrosine 155 (p-T153/Y155), was dramatically increased by the heat shock response (HSR) in human cells. HSR promotes cell survival under proteotoxic conditions by maintaining protein homeostasis and preventing protein misfolding. MEK is activated by HSR and contributes to the regulation of proteome stability. Phosphorylated TDP-43 was not associated with TDP-43 aggregation, and p-T153/Y155 remained soluble under conditions that promote protein misfolding. We found that active MEK significantly alters TDP-43-regulated splicing and that phosphomimetic substitutions at these two residues reduce binding to GU-rich RNA. Cellular imaging using a phospho-specific p-T153/Y155 antibody showed that phosphorylated TDP-43 was specifically recruited to the nucleoli, suggesting that p-T153/Y155 regulates a previously unappreciated function of TDP-43 in the processing of nucleolar-associated RNA. These findings highlight a new mechanism that regulates TDP-43 function and homeostasis through phosphorylation and, therefore, may contribute to the development of strategies to prevent TDP-43 aggregation and to uncover previously unexplored roles of TDP-43 in cell metabolism.}, }
@article {pmid28186560, year = {2017}, author = {Riar, AK and Burstein, SR and Palomo, GM and Arreguin, A and Manfredi, G and Germain, D}, title = {Sex specific activation of the ERα axis of the mitochondrial UPR (UPRmt) in the G93A-SOD1 mouse model of familial ALS.}, journal = {Human molecular genetics}, volume = {26}, number = {7}, pages = {1318-1327}, pmid = {28186560}, issn = {1460-2083}, support = {R01 NS062055/NS/NINDS NIH HHS/United States ; R01 NS084486/NS/NINDS NIH HHS/United States ; R01 NS095692/NS/NINDS NIH HHS/United States ; }, mesh = {ATP-Dependent Proteases/genetics/metabolism ; Amyotrophic Lateral Sclerosis/*genetics/metabolism/pathology ; Animals ; Disease Models, Animal ; Estrogen Receptor alpha/*genetics/metabolism ; Female ; Heat-Shock Proteins/genetics ; High-Temperature Requirement A Serine Peptidase 2 ; Humans ; Male ; Mice ; Mitochondria/*genetics/metabolism/pathology ; Mitochondrial Proteins/genetics/metabolism ; Proteasome Endopeptidase Complex/genetics/metabolism ; Serine Endopeptidases/genetics/metabolism ; Sex Characteristics ; Superoxide Dismutase/*genetics ; Unfolded Protein Response/genetics ; }, abstract = {The mitochondrial unfolded protein response (UPRmt) is a transcriptional program aimed at restoring proteostasis in mitochondria. Upregulation of mitochondrial matrix proteases and heat shock proteins was initially described. Soon thereafter, a distinct UPRmt induced by misfolded proteins in the mitochondrial intermembrane space (IMS) and mediated by the estrogen receptor alpha (ERα), was found to upregulate the proteasome and the IMS protease OMI. However, the IMS-UPRmt was never studied in a neurodegenerative disease in vivo. Thus, we investigated the IMS-UPRmt in the G93A-SOD1 mouse model of familial ALS, since mutant SOD1 is known to accumulate in the IMS of neural tissue and cause mitochondrial dysfunction. As the ERα is most active in females, we postulated that a differential involvement of the IMS-UPRmt could be linked to the longer lifespan of females in the G93A-SOD1 mouse. We found a significant sex difference in the IMS-UPRmt, because the spinal cords of female, but not male, G93A-SOD1 mice showed elevation of OMI and proteasome activity. Then, using a mouse in which G93A-SOD1 was selectively targeted to the IMS, we demonstrated that the IMS-UPRmt could be specifically initiated by mutant SOD1 localized in the IMS. Furthermore, we showed that, in the absence of ERα, G93A-SOD1 failed to activate OMI and the proteasome, confirming the ERα dependence of the response. Taken together, these results demonstrate the IMS-UPRmt activation in SOD1 familial ALS, and suggest that sex differences in the disease phenotype could be linked to differential activation of the ERα axis of the IMS-UPRmt.}, }
@article {pmid28208059, year = {2017}, author = {Shorter, J}, title = {Designer protein disaggregases to counter neurodegenerative disease.}, journal = {Current opinion in genetics &amp; development}, volume = {44}, number = {}, pages = {1-8}, pmid = {28208059}, issn = {1879-0380}, support = {R21 NS090205/NS/NINDS NIH HHS/United States ; R21 HD074510/HD/NICHD NIH HHS/United States ; DP2 OD002177/OD/NIH HHS/United States ; R01 GM099836/GM/NIGMS NIH HHS/United States ; R21 NS067354/NS/NINDS NIH HHS/United States ; }, mesh = {Alzheimer Disease/genetics ; Amyotrophic Lateral Sclerosis/genetics ; Bacteriophages/genetics ; HSP110 Heat-Shock Proteins/*genetics ; HSP40 Heat-Shock Proteins/*genetics ; HSP72 Heat-Shock Proteins/*genetics ; Heat-Shock Proteins/*genetics ; Humans ; Parkinson Disease/genetics ; Protein Aggregation, Pathological/genetics ; Protein Folding ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae Proteins/*genetics ; }, abstract = {Protein misfolding and aggregation unify several devastating neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis. There are no effective therapeutics for these disorders and none that target the reversal of the aberrant protein misfolding and aggregation that cause disease. Here, I showcase important advances to define, engineer, and apply protein disaggregases to mitigate deleterious protein misfolding and counter neurodegeneration. I focus on two exogenous protein disaggregases, Hsp104 from yeast and gene 3 protein from bacteriophages, as well as endogenous human protein disaggregases, including: (a) Hsp110, Hsp70, Hsp40, and small heat-shock proteins; (b) HtrA1; and (c) NMNAT2 and Hsp90. I suggest that protein-disaggregase modalities can be channeled to treat numerous fatal and presently incurable neurodegenerative diseases.}, }
@article {pmid28336525, year = {2017}, author = {Leal-Lasarte, MM and Franco, JM and Labrador-Garrido, A and Pozo, D and Roodveldt, C}, title = {Extracellular TDP-43 aggregates target MAPK/MAK/MRK overlapping kinase (MOK) and trigger caspase-3/IL-18 signaling in microglia.}, journal = {FASEB journal : official publication of the Federation of American Societies for Experimental Biology}, volume = {31}, number = {7}, pages = {2797-2816}, doi = {10.1096/fj.201601163R}, pmid = {28336525}, issn = {1530-6860}, mesh = {Animals ; Caspase 3/genetics/*metabolism ; Cell Survival ; Cells, Cultured ; DNA-Binding Proteins/administration &amp; dosage/*pharmacology ; Gene Expression Regulation, Enzymologic/*drug effects/physiology ; Inflammasomes/metabolism ; Inflammation/metabolism ; Interleukin-18/genetics/*metabolism ; Male ; Mice ; Microglia/*metabolism ; Mitogen-Activated Protein Kinases/genetics/*metabolism ; }, abstract = {Dysregulated microglial responses are central in neurodegenerative proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar disease (FTLD). Pathologic TDP-43, which is typically found in intracellular inclusions, is a misfolding protein with emerging roles in ALS and FTLD. Recently, TDP-43 species have been found in extracellular fluids of patients; however, the overall implications of TDP-43-mediated signaling linked to neuroinflammation are poorly understood. Our work-the first, to our knowledge, to focus on innate immunity responses to TDP-43 aggregates-shows that such species are internalized by microglia and cause abnormal mobilization of endogenous TDP-43. Exposure to TDP-43 aggregates elicited not only IL-1β, but also NLRP3-dependent and noncanonical IL-18 processing. Moreover, we report a link between TDP-43 and neuronal loss via the apoptosis-independent emerging roles of caspase-3 in neurotoxic inflammation. Our results further support the view of noncell autonomous neurodegenerative mechanisms in ALS. Remarkably, we demonstrate that TDP-43 aggregates bind to and colocalize with MAPK/MAK/MRK overlapping kinase (MOK) and show that its phosphorylation status is disrupted. Finally, we show that this TDP-43-caused activation state can be altered by exogenous Hsp27 and Hsp70 chaperones. Our study provides new insight into the immune phenotype, mechanisms, and signaling pathways that operate in microglial neurotoxic activation in ALS.-Leal-Lasarte, M. M., Franco, J. M., Labrador-Garrido, A., Pozo, D., Roodveldt, C. Extracellular TDP-43 aggregates target MAPK/MAK/MRK overlapping kinase (MOK) and trigger caspase-3/IL-18 signaling in microglia.}, }
@article {pmid28428745, year = {2017}, author = {Filareti, M and Luotti, S and Pasetto, L and Pignataro, M and Paolella, K and Messina, P and Pupillo, E and Filosto, M and Lunetta, C and Mandrioli, J and Fuda, G and Calvo, A and Chiò, A and Corbo, M and Bendotti, C and Beghi, E and Bonetto, V}, title = {Decreased Levels of Foldase and Chaperone Proteins Are Associated with an Early-Onset Amyotrophic Lateral Sclerosis.}, journal = {Frontiers in molecular neuroscience}, volume = {10}, number = {}, pages = {99}, pmid = {28428745}, issn = {1662-5099}, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by a progressive upper and lower motor neuron degeneration. One of the peculiar clinical characteristics of ALS is the wide distribution in age of onset, which is probably caused by different combinations of intrinsic and exogenous factors. We investigated whether these modifying factors are converging into common pathogenic pathways leading either to an early or a late disease onset. This would imply the identification of phenotypic biomarkers, that can distinguish the two populations of ALS patients, and of relevant pathways to consider in a therapeutic intervention. Toward this aim a differential proteomic analysis was performed in peripheral blood mononuclear cells (PBMC) from a group of 16 ALS patients with an age of onset ≤55 years and a group of 16 ALS patients with an age of onset ≥75 years, and matched healthy controls. We identified 43 differentially expressed proteins in the two groups of patients. Gene ontology analysis revealed that there was a significant enrichment in annotations associated with protein folding and response to stress. We next validated a selected number of proteins belonging to this functional group in 85 patients and 83 age- and sex-matched healthy controls using immunoassays. The results of the validation study confirmed that there was a decreased level of peptidyl-prolyl cis-trans isomerase A (also known as cyclophilin A), heat shock protein HSP 90-alpha, 78 kDa glucose-regulated protein (also known as BiP) and protein deglycase DJ-1 in PBMC of ALS patients with an early onset. Similar results were obtained in PBMC and spinal cord from two SOD1[G93A] mouse models with an early and late disease onset. This study suggests that a different ability to upregulate proteins involved in proteostasis, such as foldase and chaperone proteins, may be at the basis of a different susceptibility to ALS, putting forward the development of therapeutic approaches aiming at boosting the protein quality control system.}, }
@article {pmid28559789, year = {2017}, author = {Lackie, RE and Maciejewski, A and Ostapchenko, VG and Marques-Lopes, J and Choy, WY and Duennwald, ML and Prado, VF and Prado, MAM}, title = {The Hsp70/Hsp90 Chaperone Machinery in Neurodegenerative Diseases.}, journal = {Frontiers in neuroscience}, volume = {11}, number = {}, pages = {254}, pmid = {28559789}, issn = {1662-4548}, abstract = {The accumulation of misfolded proteins in the human brain is one of the critical features of many neurodegenerative diseases, including Alzheimer's disease (AD). Assembles of beta-amyloid (Aβ) peptide-either soluble (oligomers) or insoluble (plaques) and of tau protein, which form neurofibrillary tangles, are the major hallmarks of AD. Chaperones and co-chaperones regulate protein folding and client maturation, but they also target misfolded or aggregated proteins for refolding or for degradation, mostly by the proteasome. They form an important line of defense against misfolded proteins and are part of the cellular quality control system. The heat shock protein (Hsp) family, particularly Hsp70 and Hsp90, plays a major part in this process and it is well-known to regulate protein misfolding in a variety of diseases, including tau levels and toxicity in AD. However, the role of Hsp90 in regulating protein misfolding is not yet fully understood. For example, knockdown of Hsp90 and its co-chaperones in a Caenorhabditis elegans model of Aβ misfolding leads to increased toxicity. On the other hand, the use of Hsp90 inhibitors in AD mouse models reduces Aβ toxicity, and normalizes synaptic function. Stress-inducible phosphoprotein 1 (STI1), an intracellular co-chaperone, mediates the transfer of clients from Hsp70 to Hsp90. Importantly, STI1 has been shown to regulate aggregation of amyloid-like proteins in yeast. In addition to its intracellular function, STI1 can be secreted by diverse cell types, including astrocytes and microglia and function as a neurotrophic ligand by triggering signaling via the cellular prion protein (PrP[C]). Extracellular STI1 can prevent Aβ toxic signaling by (i) interfering with Aβ binding to PrP[C] and (ii) triggering pro-survival signaling cascades. Interestingly, decreased levels of STI1 in C. elegans can also increase toxicity in an amyloid model. In this review, we will discuss the role of intracellular and extracellular STI1 and the Hsp70/Hsp90 chaperone network in mechanisms underlying protein misfolding in neurodegenerative diseases, with particular focus on AD.}, }
@article {pmid28608264, year = {2018}, author = {Cristofani, R and Crippa, V and Vezzoli, G and Rusmini, P and Galbiati, M and Cicardi, ME and Meroni, M and Ferrari, V and Tedesco, B and Piccolella, M and Messi, E and Carra, S and Poletti, A}, title = {The small heat shock protein B8 (HSPB8) efficiently removes aggregating species of dipeptides produced in C9ORF72-related neurodegenerative diseases.}, journal = {Cell stress &amp; chaperones}, volume = {23}, number = {1}, pages = {1-12}, pmid = {28608264}, issn = {1466-1268}, support = {GGP14039//Fondazione Telethon/International ; 2014-0686//Fondazione Cariplo/International ; ALS_HSPB8//Fondazione Arisla/International ; ALS_Granulopathy//Fondazione Arisla/International ; ALS_Granulopathy//Fondazione AriSLA/International ; ALS_HSPB8//Fondazione AriSLA/International ; 16406//Association Fran&#xe7;aise contre les Myopathies/International ; piano di sviluppo UNIMI - linea B//Universit&#xe0; degli studi di Milano/International ; GR-2011-02347198//Ministero della Salute/International ; GR-2011-02347198//Ministero della Salute/International ; Cure_ALS n. 643417//Joint Programme on Neurodegenerative Diseases/International ; CureALS n. 643417//Joint Programme Neurodegenerative Disease/International ; TRANS_ALS//Fondazione regionale per la ricerca biomedica (FRRB), Regione Lombardia/International ; PRIN n. 2015LFPNMN//Ministero dell'Istruzione, dell'Universit&#xe0; e della Ricerca/International ; PRIN n. 2015LFPNMN//Ministero dell'Istruzione, dell'Universit&#xe0; e della Ricerca/International ; 537 - 2015//European Molecular Biology Organization/International ; }, mesh = {Animals ; Autophagy/drug effects ; C9orf72 Protein/*metabolism ; Cell Line, Transformed ; Dipeptides/*metabolism ; Gene Silencing ; HSP20 Heat-Shock Proteins/*metabolism ; Mice ; Motor Neurons/metabolism/*pathology ; Neurodegenerative Diseases/*metabolism/pathology ; Proteasome Inhibitors/pharmacology ; *Protein Aggregates ; Up-Regulation/drug effects ; }, abstract = {Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are two neurodegenerative diseases in which similar pathogenic mechanisms are involved. Both diseases associate to the high propensity of specific misfolded proteins, like TDP-43 or FUS, to mislocalize and aggregate. This is partly due to their intrinsic biophysical properties and partly as a consequence of failure of the neuronal protein quality control (PQC) system. Several familial ALS/FTD cases are linked to an expansion of a repeated G4C2 hexanucleotide sequence present in the C9ORF72 gene. The G4C2, which localizes in an untranslated region of the C9ORF72 transcript, drives an unconventional repeat-associated ATG-independent translation. This leads to the synthesis of five different dipeptide repeat proteins (DPRs), which are not "classical" misfolded proteins, but generate aberrant aggregation-prone unfolded conformations poorly removed by the PQC system. The DPRs accumulate into p62/SQSTM1 and ubiquitin positive inclusions. Here, we analyzed the biochemical behavior of the five DPRs in immortalized motoneurons. Our data suggest that while the DPRs are mainly processed via autophagy, this system is unable to fully clear their aggregated forms, and thus they tend to accumulate in basal conditions. Overexpression of the small heat shock protein B8 (HSPB8), which facilitates the autophagy-mediated disposal of a large variety of classical misfolded aggregation-prone proteins, significantly decreased the accumulation of most DPR insoluble species. Thus, the induction of HSPB8 might represent a valid approach to decrease DPR-mediated toxicity and maintain motoneuron viability.}, }
@article {pmid28617431, year = {2017}, author = {Corsini, S and Tortora, M and Rauti, R and Nistri, A}, title = {Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36.}, journal = {Cell death &amp; disease}, volume = {8}, number = {6}, pages = {e2881}, pmid = {28617431}, issn = {2041-4889}, mesh = {Animals ; Apoptosis Inducing Factor/*metabolism ; Brain Stem/*drug effects ; Calcium/metabolism ; Carbenoxolone/pharmacology ; Cell Death ; Connexins/*metabolism ; Down-Regulation ; Gap Junctions/metabolism ; HSP70 Heat-Shock Proteins/*metabolism ; Motor Neurons/*drug effects ; Neurons/metabolism ; Nicotine/*pharmacology ; Protein Transport ; Rats ; Rats, Wistar ; Gap Junction delta-2 Protein ; }, abstract = {Motoneuron disease including amyotrophic lateral sclerosis may be due, at an early stage, to deficit in the extracellular clearance of the excitatory transmitter glutamate. A model of glutamate-mediated excitotoxic cell death based on pharmacological inhibition of its uptake was used to investigate how activation of neuronal nicotinic receptors by nicotine may protect motoneurons. Hypoglossal motoneurons (HMs) in neonatal rat brainstem slices were exposed to the glutamate uptake blocker DL-threo-β-benzyloxyaspartate (TBOA) that evoked large Ca[2+] transients time locked among nearby HMs, whose number fell by about 30% 4 h later. As nicotine or the gap junction blocker carbenoxolone suppressed bursting, we studied connexin 36 (Cx36), which constitutes gap junctions in neurons and found it largely expressed by HMs. Cx36 was downregulated when nicotine or carbenoxolone was co-applied with TBOA. Expression of Cx36 was preferentially observed in cytosolic rather than membrane fractions after nicotine and TBOA, suggesting protein redistribution with no change in synthesis. Nicotine raised the expression of heat shock protein 70 (Hsp70), a protective factor that binds the apoptotic-inducing factor (AIF) whose nuclear translocation is a cause of cell death. TBOA increased intracellular AIF, an effect blocked by nicotine. These results indicate that activation of neuronal nicotinic receptors is an early tool for protecting motoneurons from excitotoxicity and that this process is carried out via the combined decrease in Cx36 activity, overexpression of Hsp70 and fall in AIF translocation. Thus, retarding or inhibiting HM death may be experimentally achieved by targeting one of these processes leading to motoneuron death.}, }
@article {pmid28651026, year = {2017}, author = {Karkowska-Kuleta, J and Zajac, D and Bras, G and Bochenska, O and Rapala-Kozik, M and Kozik, A}, title = {Binding of human plasminogen and high-molecular-mass kininogen by cell surface-exposed proteins of Candida parapsilosis.}, journal = {Acta biochimica Polonica}, volume = {64}, number = {3}, pages = {391-400}, doi = {10.18388/abp.2017_1609}, pmid = {28651026}, issn = {1734-154X}, mesh = {Binding, Competitive ; Candida parapsilosis/*metabolism/pathogenicity ; Cell Wall/metabolism ; Chromatography, Affinity ; Fungal Proteins/isolation &amp; purification/*metabolism ; Host-Pathogen Interactions ; Humans ; Hyphae/metabolism ; Kinetics ; Kininogen, High-Molecular-Weight/*metabolism ; Kininogens/metabolism ; Plasminogen/*metabolism ; Thermodynamics ; }, abstract = {Pathogenic microbes can recruit to their cell surface human proteins that are components of important proteolytic cascades involved in coagulation, fibrinolysis and innate immune response. Once located at the bacterial or fungal surface, such deployed proteins might be utilized by pathogens to facilitate invasion and dissemination within the host organism by interfering with functionality of these systems or by exploiting specific activity of the bound enzymes. Aim of the study presented here was to characterize this phenomenon in Candida parapsilosis (Ashford) Langeron et Talice - an important causative agent of systemic fungal infections (candidiases and candidemias) in humans. We have investigated the interactions of fungal surface-exposed proteins with plasminogen (HPG) and high-molecular-mass kininogen (HK) - the crucial components of human fibrinolytic system and proinflammatory/procoagulant contact-activated kinin-forming system, respectively. After confirming ability of the fungal surface-exposed proteins to bind HPG and HK, four of them - two agglutinin-like sequence (Als) proteins CPAR2_404780 and CPAR2_404800, a heat shock protein Ssa2 and a moonlighting protein 6-phosphogluconate dehydrogenase 1 - were purified using ion-exchange chromatography, gel filtration and chromatofocusing. Then, their affinities to HPG and HK were characterized with surface plasmon resonance measurements. The determined dissociation constants for the investigated protein-protein complexes were within a 10[-7] M order for the HPG binding and in a range of 10[-8]-10[-9] M for the HK binding. Detailed characterization of adsorption of these two important plasma proteins on the fungal cell surface may help to increase our understanding of molecular mechanisms of C. parapsilosis-dependent candidiasis.}, }
@article {pmid28680390, year = {2017}, author = {Rusmini, P and Cristofani, R and Galbiati, M and Cicardi, ME and Meroni, M and Ferrari, V and Vezzoli, G and Tedesco, B and Messi, E and Piccolella, M and Carra, S and Crippa, V and Poletti, A}, title = {The Role of the Heat Shock Protein B8 (HSPB8) in Motoneuron Diseases.}, journal = {Frontiers in molecular neuroscience}, volume = {10}, number = {}, pages = {176}, pmid = {28680390}, issn = {1662-5099}, abstract = {Amyotrophic lateral sclerosis (ALS) and spinal and bulbar muscular atrophy (SBMA) are two motoneuron diseases (MNDs) characterized by aberrant protein behavior in affected cells. In familial ALS (fALS) and in SBMA specific gene mutations lead to the production of neurotoxic proteins or peptides prone to misfold, which then accumulate in form of aggregates. Notably, some of these proteins accumulate into aggregates also in sporadic ALS (sALS) even if not mutated. To prevent proteotoxic stresses detrimental to cells, misfolded and/or aggregated proteins must be rapidly removed by the protein quality control (PQC) system. The small heat shock protein B8 (HSPB8) is a chaperone induced by harmful events, like proteasome inhibition. HSPB8 is expressed both in motoneuron and muscle cells, which are both targets of misfolded protein toxicity in MNDs. In ALS mice models, in presence of the mutant proteins, HSPB8 is upregulated both in spinal cord and muscle. HSPB8 interacts with the HSP70 co-chaperone BAG3 and enhances the degradation of misfolded proteins linked to sALS, or causative of fALS and of SBMA. HSPB8 acts by facilitating autophagy, thereby preventing misfolded protein accumulation in affected cells. BAG3 and BAG1 compete for HSP70-bound clients and target them for disposal to the autophagy or proteasome, respectively. Enhancing the selective targeting of misfolded proteins by HSPB8-BAG3-HSP70 to autophagy may also decrease their delivery to the proteasome by the BAG1-HSP70 complex, thereby limiting possible proteasome overwhelming. Thus, approaches aimed at potentiating HSPB8-BAG3 may contribute to the maintenance of proteostasis and may delay MNDs progression.}, }
@article {pmid28797631, year = {2017}, author = {Heilman, PL and Song, S and Miranda, CJ and Meyer, K and Srivastava, AK and Knapp, A and Wier, CG and Kaspar, BK and Kolb, SJ}, title = {HSPB1 mutations causing hereditary neuropathy in humans disrupt non-cell autonomous protection of motor neurons.}, journal = {Experimental neurology}, volume = {297}, number = {}, pages = {101-109}, pmid = {28797631}, issn = {1090-2430}, support = {K08 NS067282/NS/NINDS NIH HHS/United States ; P30 NS104177/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Astrocytes/pathology/*physiology ; Cell Survival/physiology ; Charcot-Marie-Tooth Disease/*genetics/pathology ; Coculture Techniques ; Heat-Shock Proteins/*genetics ; Humans ; Mice ; Mice, Transgenic ; Molecular Chaperones ; Motor Neurons/pathology/*physiology ; Mutation/*genetics ; Neoplasm Proteins/*genetics ; Neuroglia/pathology/*physiology ; }, abstract = {Heat shock protein beta-1 (HSPB1), is a ubiquitously expressed, multifunctional protein chaperone. Mutations in HSPB1 result in the development of a late-onset, distal hereditary motor neuropathy type II (dHMN) and axonal Charcot-Marie Tooth disease with sensory involvement (CMT2F). The functional consequences of HSPB1 mutations associated with hereditary neuropathy are unknown. HSPB1 also displays neuroprotective properties in many neuronal disease models, including the motor neuron disease amyotrophic lateral sclerosis (ALS). HSPB1 is upregulated in SOD1-ALS animal models during disease progression, predominately in glial cells. Glial cells are known to contribute to motor neuron loss in ALS through a non-cell autonomous mechanism. In this study, we examined the non-cell autonomous role of wild type and mutant HSPB1 in an astrocyte-motor neuron co-culture model system of ALS. Astrocyte-specific overexpression of wild type HSPB1 was sufficient to attenuate SOD1(G93A) astrocyte-mediated toxicity in motor neurons, whereas, overexpression of mutHSPB1 failed to ameliorate motor neuron toxicity. Expression of a phosphomimetic HSPB1 mutant in SOD1(G93A) astrocytes also reduced toxicity to motor neurons, suggesting that phosphorylation may contribute to HSPB1 mediated-neuroprotection. These data provide evidence that astrocytic HSPB1 expression may play a central role in motor neuron health and maintenance.}, }
@article {pmid28852778, year = {2018}, author = {Lee, A and Rayner, SL and Gwee, SSL and De Luca, A and Shahheydari, H and Sundaramoorthy, V and Ragagnin, A and Morsch, M and Radford, R and Galper, J and Freckleton, S and Shi, B and Walker, AK and Don, EK and Cole, NJ and Yang, S and Williams, KL and Yerbury, JJ and Blair, IP and Atkin, JD and Molloy, MP and Chung, RS}, title = {Pathogenic mutation in the ALS/FTD gene, CCNF, causes elevated Lys48-linked ubiquitylation and defective autophagy.}, journal = {Cellular and molecular life sciences : CMLS}, volume = {75}, number = {2}, pages = {335-354}, pmid = {28852778}, issn = {1420-9071}, support = {APP1036835//National Health and Medical Research Council/International ; APP1107644//National Health and Medical Research Council/International ; APP1030513//National Health and Medical Research Council/International ; APP1095215//National Health and Medical Research Council/International ; GIA1510//Motor Neurone Disease Research Institute of Australia/International ; GIA1628//Motor Neurone Disease Research Institute of Australia/International ; GIA1715//Motor Neurone Disease Research Institute of Australia/International ; }, mesh = {Amyotrophic Lateral Sclerosis/complications/*genetics ; Autophagy/*genetics ; Cells, Cultured ; Cyclins/*genetics ; Frontotemporal Dementia/complications/*genetics ; HEK293 Cells ; Humans ; Lysine/metabolism ; Mutation, Missense/physiology ; Ubiquitination/*genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are fatal neurodegenerative disorders that have common molecular and pathogenic characteristics, such as aberrant accumulation and ubiquitylation of TDP-43; however, the mechanisms that drive this process remain poorly understood. We have recently identified CCNF mutations in familial and sporadic ALS and FTD patients. CCNF encodes cyclin F, a component of an E3 ubiquitin-protein ligase (SCF[cyclin F]) complex that is responsible for ubiquitylating proteins for degradation by the ubiquitin-proteasome system. In this study, we examined the ALS/FTD-causing p.Ser621Gly (p.S621G) mutation in cyclin F and its effect upon downstream Lys48-specific ubiquitylation in transfected Neuro-2A and SH-SY5Y cells. Expression of mutant cyclin F[S621G] caused increased Lys48-specific ubiquitylation of proteins in neuronal cells compared to cyclin F[WT]. Proteomic analysis of immunoprecipitated Lys48-ubiquitylated proteins from mutant cyclin F[S621G]-expressing cells identified proteins that clustered within the autophagy pathway, including sequestosome-1 (p62/SQSTM1), heat shock proteins, and chaperonin complex components. Examination of autophagy markers p62, LC3, and lysosome-associated membrane protein 2 (Lamp2) in cells expressing mutant cyclin F[S621G] revealed defects in the autophagy pathway specifically resulting in impairment in autophagosomal-lysosome fusion. This finding highlights a potential mechanism by which cyclin F interacts with p62, the receptor responsible for transporting ubiquitylated substrates for autophagic degradation. These findings demonstrate that ALS/FTD-causing mutant cyclin F[S621G] disrupts Lys48-specific ubiquitylation, leading to accumulation of substrates and defects in the autophagic machinery. This study also demonstrates that a single missense mutation in cyclin F causes hyper-ubiquitylation of proteins that can indirectly impair the autophagy degradation pathway, which is implicated in ALS pathogenesis.}, }
@article {pmid28923065, year = {2017}, author = {San Gil, R and Ooi, L and Yerbury, JJ and Ecroyd, H}, title = {The heat shock response in neurons and astroglia and its role in neurodegenerative diseases.}, journal = {Molecular neurodegeneration}, volume = {12}, number = {1}, pages = {65}, pmid = {28923065}, issn = {1750-1326}, mesh = {Animals ; Astrocytes/*metabolism/pathology ; Heat-Shock Response/*physiology ; Humans ; Neurodegenerative Diseases/*metabolism/pathology ; Neurons/*metabolism/pathology ; }, abstract = {Protein inclusions are a predominant molecular pathology found in numerous neurodegenerative diseases, including amyotrophic lateral sclerosis and Huntington's disease. Protein inclusions form in discrete areas of the brain characteristic to the type of neurodegenerative disease, and coincide with the death of neurons in that region (e.g. spinal cord motor neurons in amyotrophic lateral sclerosis). This suggests that the process of protein misfolding leading to inclusion formation is neurotoxic, and that cell-autonomous and non-cell autonomous mechanisms that maintain protein homeostasis (proteostasis) can, at times, be insufficient to prevent protein inclusion formation in the central nervous system. The heat shock response is a pro-survival pathway induced under conditions of cellular stress that acts to maintain proteostasis through the up-regulation of heat shock proteins, a superfamily of molecular chaperones, other co-chaperones and mitotic regulators. The kinetics and magnitude of the heat shock response varies in a stress- and cell-type dependent manner. It remains to be determined if and/or how the heat shock response is activated in the different cell-types that comprise the central nervous system (e.g. neurons and astroglia) in response to protein misfolding events that precede cellular dysfunctions in neurodegenerative diseases. This is particularly relevant considering emerging evidence demonstrating the non-cell autonomous nature of amyotrophic lateral sclerosis and Huntington's disease (and other neurodegenerative diseases) and the destructive role of astroglia in disease progression. This review highlights the complexity of heat shock response activation and addresses whether neurons and glia sense and respond to protein misfolding and aggregation associated with neurodegenerative diseases, in particular Huntington's disease and amyotrophic lateral sclerosis, by inducing a pro-survival heat shock response.}, }
@article {pmid28943839, year = {2017}, author = {Kalmar, B and Greensmith, L}, title = {Cellular Chaperones As Therapeutic Targets in ALS to Restore Protein Homeostasis and Improve Cellular Function.}, journal = {Frontiers in molecular neuroscience}, volume = {10}, number = {}, pages = {251}, pmid = {28943839}, issn = {1662-5099}, abstract = {Heat shock proteins (Hsps) are ubiquitously expressed chaperone proteins that enable cells to cope with environmental stresses that cause misfolding and denaturation of proteins. With aging this protein quality control machinery becomes less effective, reducing the ability of cells to cope with damaging environmental stresses and disease-causing mutations. In neurodegenerative disorders such as Amyotrophic Lateral Sclerosis (ALS), such mutations are known to result in protein misfolding, which in turn results in the formation of intracellular aggregates cellular dysfunction and eventual neuronal death. The exact cellular pathology of ALS and other neurodegenerative diseases has been elusive and thus, hindering the development of effective therapies. However, a common scheme has emerged across these "protein misfolding" disorders, in that the mechanism of disease involves one or more aspects of proteostasis; from DNA transcription, RNA translation, to protein folding, transport and degradation via proteosomal and autophagic pathways. Interestingly, members of the Hsp family are involved in each of these steps facilitating normal protein folding, regulating the rate of protein synthesis and degradation. In this short review we summarize the evidence that suggests that ALS is a disease of protein dyshomeostasis in which Hsps may play a key role. Overwhelming evidence now indicates that enabling protein homeostasis to cope with disease-causing mutations might be a successful therapeutic strategy in ALS, as well as other neurodegenerative diseases. Novel small molecule co-inducers of Hsps appear to be able to achieve this aim. Arimoclomol, a hydroxylamine derivative, has shown promising results in cellular and animal models of ALS, as well as other protein misfolding diseases such as Inclusion Body Myositis (IBM). Initial clinical investigations of Arimoclomol have shown promising results. Therefore, it is possible that the long series of unsuccessful clinical trials for ALS may soon be reversed, as optimal targeting of proteostasis in ALS may now be possible, and may deliver clinical benefit to patients.}, }
@article {pmid28969372, year = {2017}, author = {Adriaenssens, E and Geuens, T and Baets, J and Echaniz-Laguna, A and Timmerman, V}, title = {Novel insights in the disease biology of mutant small heat shock proteins in neuromuscular diseases.}, journal = {Brain : a journal of neurology}, volume = {140}, number = {10}, pages = {2541-2549}, doi = {10.1093/brain/awx187}, pmid = {28969372}, issn = {1460-2156}, mesh = {Aged ; Aged, 80 and over ; HSP27 Heat-Shock Proteins/*genetics ; Heat-Shock Proteins/*genetics ; Humans ; Male ; Models, Molecular ; Molecular Chaperones ; Mutation/*genetics ; Neuromuscular Diseases/*genetics ; Protein Serine-Threonine Kinases/*genetics ; }, abstract = {Small heat shock proteins are molecular chaperones that exert diverse cellular functions. To date, mutations in the coding regions of HSPB1 (Hsp27) and HSPB8 (Hsp22) were reported to cause distal hereditary motor neuropathy and Charcot-Marie-Tooth disease. Recently, the clinical spectrum of HSPB1 and HSPB8 mutations was expanded to also include myopathies. Here we provide an update on the molecular genetics and biology of small heat shock protein mutations in neuromuscular diseases.}, }
@article {pmid29035885, year = {2017}, author = {Lo Bello, M and Di Fini, F and Notaro, A and Spataro, R and Conforti, FL and La Bella, V}, title = {ALS-Related Mutant FUS Protein Is Mislocalized to Cytoplasm and Is Recruited into Stress Granules of Fibroblasts from Asymptomatic FUS P525L Mutation Carriers.}, journal = {Neuro-degenerative diseases}, volume = {17}, number = {6}, pages = {292-303}, doi = {10.1159/000480085}, pmid = {29035885}, issn = {1660-2862}, mesh = {Amyotrophic Lateral Sclerosis/*genetics/*pathology/physiopathology ; Cell Nucleus/metabolism ; Cells, Cultured ; Cytoplasmic Granules/metabolism ; Female ; Fibroblasts/*metabolism ; Follow-Up Studies ; Humans ; Leucine/genetics ; Male ; Mutation/*genetics ; Neural Conduction/genetics ; Proline/genetics ; Protein Transport/*genetics ; RNA-Binding Protein FUS/*genetics ; Skin/cytology ; Subcellular Fractions/metabolism/pathology ; Time Factors ; Tubulin/metabolism ; }, abstract = {BACKGROUND: Amyotrophic lateral sclerosis (ALS) shows a strong genetic basis, with SOD1, FUS, TARDBP, and C9ORF72 being the genes most frequently involved. This has allowed identification of asymptomatic mutation carriers, which may be of help in understanding the molecular changes preceding disease onset.<br><br>OBJECTIVES: We studied the cellular expression of FUS protein and the effect of heat-shock- and dithiothreitol-induced stress in fibroblasts from FUS P525L mutation carriers, healthy controls, and patients with sporadic ALS.<br><br>METHODS: Western blots and immunocytochemistry were performed to study the subcellular localization of FUS protein. Control and stressed cells were double stained with FUS and the stress marker TIA-R.<br><br>RESULTS: Fibroblasts from healthy controls and sporadic ALS cases showed a prominent nuclear FUS expression. In the 2 FUS P525L mutation carriers, instead, most cells showed a protein localization in both nucleus and cytoplasm, or exclusively in the cytoplasm. Stress prompted the formation of cytoplasmic granules in all subjects and in sporadic ALS FUS mislocalization to the cytoplasm. Cytoplasmic FUS was recruited into stress granules, which showed a time-dependent decrease in all subjects. However, in the FUS P525L fibroblasts, the granules persisted longer, and they were more numerous than those detected in the cells from controls and sporadic ALS patients.<br><br>CONCLUSIONS: We show that in fibroblasts of FUS P525L mutation carriers, FUS mislocalized to the cytoplasm where it redistributed into stress granules with likely a dose effect, i.e. a higher number of cells with granules, which persist longer, than in controls and ALS cases. These data represent an early molecular change occurring before ALS onset, suggesting a transient preaggregative state.}, }
@article {pmid29153328, year = {2017}, author = {Berson, A and Sartoris, A and Nativio, R and Van Deerlin, V and Toledo, JB and Porta, S and Liu, S and Chung, CY and Garcia, BA and Lee, VM and Trojanowski, JQ and Johnson, FB and Berger, SL and Bonini, NM}, title = {TDP-43 Promotes Neurodegeneration by Impairing Chromatin Remodeling.}, journal = {Current biology : CB}, volume = {27}, number = {23}, pages = {3579-3590.e6}, pmid = {29153328}, issn = {1879-0445}, support = {P30 AG010124/AG/NIA NIH HHS/United States ; P01 AG017586/AG/NIA NIH HHS/United States ; R01 NS078283/NS/NINDS NIH HHS/United States ; P01 CA196539/CA/NCI NIH HHS/United States ; P01 AG031862/AG/NIA NIH HHS/United States ; P30 ES013508/ES/NIEHS NIH HHS/United States ; R01 GM110174/GM/NIGMS NIH HHS/United States ; R35 NS097275/NS/NINDS NIH HHS/United States ; F32 NS084667/NS/NINDS NIH HHS/United States ; }, mesh = {Adult ; Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/*genetics/metabolism/physiopathology ; Animals ; *Chromatin Assembly and Disassembly ; DNA-Binding Proteins/*genetics/metabolism ; Disease Models, Animal ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster/*genetics/metabolism ; Frontotemporal Dementia/*genetics/metabolism/physiopathology ; HEK293 Cells ; Heat-Shock Proteins/metabolism ; Humans ; Male ; Middle Aged ; }, abstract = {Regulation of chromatin structure is critical for brain development and function. However, the involvement of chromatin dynamics in neurodegeneration is less well understood. Here we find, launching from Drosophila models of amyotrophic lateral sclerosis and frontotemporal dementia, that TDP-43 impairs the induction of multiple key stress genes required to protect from disease by reducing the recruitment of the chromatin remodeler Chd1 to chromatin. Chd1 depletion robustly enhances TDP-43-mediated neurodegeneration and promotes the formation of stress granules. Conversely, upregulation of Chd1 restores nucleosomal dynamics, promotes normal induction of protective stress genes, and rescues stress sensitivity of TDP-43-expressing animals. TDP-43-mediated impairments are conserved in mammalian cells, and, importantly, the human ortholog CHD2 physically interacts with TDP-43 and is strikingly reduced in level in temporal cortex of human patient tissue. These findings indicate that TDP-43-mediated neurodegeneration causes impaired chromatin dynamics that prevents appropriate expression of protective genes through compromised function of the chromatin remodeler Chd1/CHD2. Enhancing chromatin dynamics may be a treatment approach to amyotrophic lateral scleorosis (ALS)/frontotemporal dementia (FTD).}, }
@article {pmid29186496, year = {2018}, author = {Braak, H and Del Tredici, K}, title = {Anterior Cingulate Cortex TDP-43 Pathology in Sporadic Amyotrophic Lateral Sclerosis.}, journal = {Journal of neuropathology and experimental neurology}, volume = {77}, number = {1}, pages = {74-83}, doi = {10.1093/jnen/nlx104}, pmid = {29186496}, issn = {1554-6578}, mesh = {Adult ; Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/*metabolism/pathology ; DNA-Binding Proteins/*metabolism ; Female ; Gyrus Cinguli/*metabolism/pathology ; Humans ; Male ; Middle Aged ; Neurons/metabolism/pathology ; }, abstract = {Neuronal constituents of the human anterior cingulate cortex displayed morphological changes related to the 43-kDa transactive response DNA-binding protein (TDP-43) in advanced pathological stages of sporadic amyotrophic lateral sclerosis (sALS). By using nonphosphorylation-dependent TDP-43 immunocytochemistry, it was seen that the changes in susceptible pyramidal cells of the superficial cellular layers II-IIIab differed from those in the deep layers IIIc-Vb: A complete loss of nuclear TDP-43 expression (i.e. nuclear clearing) in the small projection neurons of layers II-IIIab was consistently accompanied by the development of somatic skein-like TDP-43-immunopositive inclusions. In contrast, in the large pyramidal cells of layers IIIc-Vb and von Economo neurons of layer Vb, skein-like inclusions were lacking or, when aggregated TDP-43 was present, the aggregates presented as dash-like TDP-43-immunopositive particles in the vicinity of the cell nucleus. The cytoskeleton of projection neurons in layers II-IIIab is neurofilament-sparse in contrast to that of the large neurons in layers IIIc-Vb, which are rich in neurofilaments and also heat shock proteins that function as their molecular chaperones. The disparities between the two neuronal populations may contribute to the two differing immunocytochemical profiles reported here. Some implications of the findings for the pathogenesis and progression of TDP-43 pathology in sALS are discussed.}, }
@article {pmid29203718, year = {2018}, author = {Zarouchlioti, C and Parfitt, DA and Li, W and Gittings, LM and Cheetham, ME}, title = {DNAJ Proteins in neurodegeneration: essential and protective factors.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {373}, number = {1738}, pages = {}, pmid = {29203718}, issn = {1471-2970}, support = {205041/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; CHEETHAM/OCT15/881-792/MNDA_/Motor Neurone Disease Association/United Kingdom ; MR/N004434/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Fetal Proteins/*genetics/metabolism ; HSP40 Heat-Shock Proteins/*genetics/metabolism ; Humans ; Mice ; Molecular Chaperones/*genetics/metabolism ; Neurodegenerative Diseases/*genetics ; Protein Folding ; Rats ; }, abstract = {Maintenance of protein homeostasis is vitally important in post-mitotic cells, particularly neurons. Neurodegenerative diseases such as polyglutamine expansion disorders-like Huntington's disease or spinocerebellar ataxia (SCA), Alzheimer's disease, fronto-temporal dementia (FTD), amyotrophic lateral sclerosis (ALS) and Parkinson's disease-are often characterized by the presence of inclusions of aggregated protein. Neurons contain complex protein networks dedicated to protein quality control and maintaining protein homeostasis, or proteostasis. Molecular chaperones are a class of proteins with prominent roles in maintaining proteostasis, which act to bind and shield hydrophobic regions of nascent or misfolded proteins while allowing correct folding, conformational changes and enabling quality control. There are many different families of molecular chaperones with multiple functions in proteostasis. The DNAJ family of molecular chaperones is the largest chaperone family and is defined by the J-domain, which regulates the function of HSP70 chaperones. DNAJ proteins can also have multiple other protein domains such as ubiquitin-interacting motifs or clathrin-binding domains leading to diverse and specific roles in the cell, including targeting client proteins for degradation via the proteasome, chaperone-mediated autophagy and uncoating clathrin-coated vesicles. DNAJ proteins can also contain ER-signal peptides or mitochondrial leader sequences, targeting them to specific organelles in the cell. In this review, we discuss the multiple roles of DNAJ proteins and in particular focus on the role of DNAJ proteins in protecting against neurodegenerative diseases caused by misfolded proteins. We also discuss the role of DNAJ proteins as direct causes of inherited neurodegeneration via mutations in DNAJ family genes.This article is part of the theme issue 'Heat shock proteins as modulators and therapeutic targets of chronic disease: an integrated perspective'.}, }
@article {pmid29312526, year = {2017}, author = {Maguire, G}, title = {Amyotrophic lateral sclerosis as a protein level, non-genomic disease: Therapy with S2RM exosome released molecules.}, journal = {World journal of stem cells}, volume = {9}, number = {11}, pages = {187-202}, pmid = {29312526}, issn = {1948-0210}, abstract = {Amyotrophic lateral sclerosis (ALS) is a rapidly progressing neurodegenerative disease that leads to death. No effective treatments are currently available. Based on data from epidemiological, etiological, laboratory, and clinical studies, I offer a new way of thinking about ALS and its treatment. This paper describes a host of extrinsic factors, including the exposome, that disrupt the extracellular matrix and protein function such that a spreading, prion-like disease leads to neurodegeneration in the motor tracts. A treatment regimen is described using the stem cell released molecules from a number of types of adult stem cells to provide tissue dependent molecules that restore homeostasis, including proteostasis, in the ALS patient. Because stem cells themselves as a therapeutic are cumbersome and expensive, and when implanted in a host cause aging of the host tissue and often fail to engraft or remain viable, only the S2RM molecules are used. Rebuilding of the extracellular matrix and repair of the dysfunctional proteins in the ALS patient ensues.}, }
@article {pmid29361800, year = {2018}, author = {Penke, B and Bogár, F and Crul, T and Sántha, M and Tóth, ME and Vígh, L}, title = {Heat Shock Proteins and Autophagy Pathways in Neuroprotection: from Molecular Bases to Pharmacological Interventions.}, journal = {International journal of molecular sciences}, volume = {19}, number = {1}, pages = {}, pmid = {29361800}, issn = {1422-0067}, mesh = {Animals ; Autophagy/*genetics ; Endoplasmic Reticulum/metabolism ; Endoplasmic Reticulum Stress ; Endosomes/metabolism ; Heat-Shock Proteins/*genetics/*metabolism ; Humans ; Lysosomes/metabolism ; Neurodegenerative Diseases/genetics/metabolism/prevention &amp; control/therapy ; Neurons/metabolism ; Neuroprotection/*genetics ; Proteasome Endopeptidase Complex/metabolism ; Proteolysis ; *Signal Transduction ; Ubiquitin/metabolism ; Unfolded Protein Response ; }, abstract = {Neurodegenerative diseases (NDDs) such as Alzheimer's disease, Parkinson's disease and Huntington's disease (HD), amyotrophic lateral sclerosis, and prion diseases are all characterized by the accumulation of protein aggregates (amyloids) into inclusions and/or plaques. The ubiquitous presence of amyloids in NDDs suggests the involvement of disturbed protein homeostasis (proteostasis) in the underlying pathomechanisms. This review summarizes specific mechanisms that maintain proteostasis, including molecular chaperons, the ubiquitin-proteasome system (UPS), endoplasmic reticulum associated degradation (ERAD), and different autophagic pathways (chaperon mediated-, micro-, and macro-autophagy). The role of heat shock proteins (Hsps) in cellular quality control and degradation of pathogenic proteins is reviewed. Finally, putative therapeutic strategies for efficient removal of cytotoxic proteins from neurons and design of new therapeutic targets against the progression of NDDs are discussed.}, }
@article {pmid29367439, year = {2018}, author = {Benatar, M and Wuu, J and Andersen, PM and Atassi, N and David, W and Cudkowicz, M and Schoenfeld, D}, title = {Randomized, double-blind, placebo-controlled trial of arimoclomol in rapidly progressive SOD1 ALS.}, journal = {Neurology}, volume = {90}, number = {7}, pages = {e565-e574}, pmid = {29367439}, issn = {1526-632X}, support = {R01 FD003517/FD/FDA HHS/United States ; }, mesh = {Adult ; Aged ; Amyotrophic Lateral Sclerosis/*drug therapy/genetics ; Disease Progression ; Double-Blind Method ; Female ; Humans ; Hydroxylamines/adverse effects/*therapeutic use ; Male ; Middle Aged ; Neuroprotective Agents/adverse effects/*therapeutic use ; Severity of Illness Index ; Superoxide Dismutase-1/genetics ; Survival Analysis ; Treatment Outcome ; }, abstract = {OBJECTIVE: To examine the safety and tolerability as well as the preliminary efficacy of arimoclomol, a heat shock protein co-inducer that promotes nascent protein folding, in patients with rapidly progressive SOD1 amyotrophic lateral sclerosis (ALS).<br><br>METHODS: This was a double-blind, placebo-controlled trial in which patients with rapidly progressive SOD1-mutant ALS were randomized 1:1 to receive arimoclomol 200 mg tid or matching placebo for up to 12 months. Study procedures were performed using a mix of in-person and remote assessments. Primary outcome was safety and tolerability. Secondary outcome was efficacy, with survival as the principal measure. Additional efficacy measures were the rates of decline of the Revised ALS Functional Rating Scale (ALSFRS-R) and percent predicted forced expiratory volume in 6 seconds (FEV6), and the Combined Assessment of Function and Survival (CAFS).<br><br>RESULTS: Thirty-eight participants were randomized. Thirty-six (19 placebo, 17 arimoclomol) were included in the prespecified intent-to-treat analysis. Apart from respiratory function, groups were generally well-balanced at baseline. Adverse events occurred infrequently, and were usually mild and deemed unlikely or not related to study drug. Adjusting for riluzole and baseline ALSFRS-R, survival favored arimoclomol with a hazard ratio of 0.77 (95% confidence interval [CI] 0.32-1.80). ALSFRS-R and FEV6 declined more slowly in the arimoclomol group, with treatment differences of 0.5 point/month (95% CI -0.63 to 1.63) and 1.24 percent predicted/month (95% CI -2.77 to 5.25), respectively, and the CAFS similarly favored arimoclomol.<br><br>CONCLUSIONS: This study provides Class II evidence that arimoclomol is safe and well-tolerated at a dosage of 200 mg tid for up to 12 months. Although not powered for therapeutic effect, the consistency of results across the range of prespecified efficacy outcome measures suggests a possible therapeutic benefit of arimoclomol.<br><br>CLINICALTRIALSGOV IDENTIFIER: NCT00706147.<br><br>CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that arimoclomol is safe and well-tolerated at a dosage of 200 mg tid for up to 12 months. The study lacked the precision to conclude, or to exclude, an important therapeutic benefit of arimoclomol.}, }
@article {pmid29373831, year = {2018}, author = {Markmiller, S and Soltanieh, S and Server, KL and Mak, R and Jin, W and Fang, MY and Luo, EC and Krach, F and Yang, D and Sen, A and Fulzele, A and Wozniak, JM and Gonzalez, DJ and Kankel, MW and Gao, FB and Bennett, EJ and Lécuyer, E and Yeo, GW}, title = {Context-Dependent and Disease-Specific Diversity in Protein Interactions within Stress Granules.}, journal = {Cell}, volume = {172}, number = {3}, pages = {590-604.e13}, pmid = {29373831}, issn = {1097-4172}, support = {RF1 NS101986/NS/NINDS NIH HHS/United States ; R01 NS066586/NS/NINDS NIH HHS/United States ; R01 HG004659/HG/NHGRI NIH HHS/United States ; R01 NS101986/NS/NINDS NIH HHS/United States ; R01 NS103172/NS/NINDS NIH HHS/United States ; T32 GM007198/GM/NIGMS NIH HHS/United States ; DP2 GM119132/GM/NIGMS NIH HHS/United States ; T32 GM007752/GM/NIGMS NIH HHS/United States ; }, mesh = {Amyotrophic Lateral Sclerosis/*metabolism ; Animals ; Cytoplasmic Granules/*metabolism ; Drosophila melanogaster ; HEK293 Cells ; HeLa Cells ; Humans ; Neurons/metabolism ; *Protein Interaction Maps ; Protein Transport ; Ribonucleoproteins/*metabolism ; *Stress, Physiological ; }, abstract = {Stress granules (SGs) are transient ribonucleoprotein (RNP) aggregates that form during cellular stress and are increasingly implicated in human neurodegeneration. To study the proteome and compositional diversity of SGs in different cell types and in the context of neurodegeneration-linked mutations, we used ascorbate peroxidase (APEX) proximity labeling, mass spectrometry, and immunofluorescence to identify ∼150 previously unknown human SG components. A highly integrated, pre-existing SG protein interaction network in unstressed cells facilitates rapid coalescence into larger SGs. Approximately 20% of SG diversity is stress or cell-type dependent, with neuronal SGs displaying a particularly complex repertoire of proteins enriched in chaperones and autophagy factors. Strengthening the link between SGs and neurodegeneration, we demonstrate aberrant dynamics, composition, and subcellular distribution of SGs in cells from amyotrophic lateral sclerosis (ALS) patients. Using three Drosophila ALS/FTD models, we identify SG-associated modifiers of neurotoxicity in vivo. Altogether, our results highlight SG proteins as central to understanding and ultimately targeting neurodegeneration.}, }
@article {pmid29399337, year = {2018}, author = {Abdanipour, A and Jafari Anarkooli, I and Shokri, S and Ghorbanlou, M and Bayati, V and Nejatbakhsh, R}, title = {Neuroprotective effects of selegiline on rat neural stem cells treated with hydrogen peroxide.}, journal = {Biomedical reports}, volume = {8}, number = {1}, pages = {41-46}, pmid = {29399337}, issn = {2049-9434}, abstract = {Oxidative stress and reactive oxygen species generation have been implicated in the pathogenesis of several neurological disorders including Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis and multiple sclerosis. In the present study, the neuroprotective effects of selegiline against hydrogen peroxide-induced oxidative stress in hippocampus-derived neural stem cells (NSCs) were evaluated. NSCs isolated from neonatal Wistar rats were pretreated with different doses of selegiline for 48 h and then exposed to 125 µM H2O2 for 30 min. Using MTT and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays, acridine orange/ethidium bromide staining and reverse transcription-quantitative polymerase chain reaction, the effects of selegiline on cell survival, apoptosis and the expression of B-cell lymphoma 2 (Bcl-2) and heat shock protein 4 (Hspa4) in pretreated stem cells were assessed compared with a control group lacking pretreatment. The results indicated that the viability of cells pretreated with 20 µM selegiline was significantly increased compared with the control group (P<0.05). Additionally, 20 µM selegiline increased the mRNA expression of Bcl-2 and Hspa4 (P<0.05 vs. control) and suppressed oxidative stress-induced cell death (apoptosis and necrosis; P<0.05 vs. control and 10 µM groups). From these findings, it was concluded that selegiline may be a therapeutic candidate for the treatment of neurological diseases mediated by oxidative stress.}, }
@article {pmid29416499, year = {2018}, author = {Bond, L and Bernhardt, K and Madria, P and Sorrentino, K and Scelsi, H and Mitchell, CS}, title = {A Metadata Analysis of Oxidative Stress Etiology in Preclinical Amyotrophic Lateral Sclerosis: Benefits of Antioxidant Therapy.}, journal = {Frontiers in neuroscience}, volume = {12}, number = {}, pages = {10}, pmid = {29416499}, issn = {1662-4548}, support = {R03 NS098228/NS/NINDS NIH HHS/United States ; }, abstract = {Oxidative stress, induced by an imbalance of free radicals, incites neurodegeneration in Amyotrophic Lateral Sclerosis (ALS). In fact, a mutation in antioxidant enzyme superoxide dismutase 1 (SOD1) accounts for 20% of familial ALS cases. However, the variance among individual studies examining ALS oxidative stress clouds corresponding conclusions. Therefore, we construct a comprehensive, temporal view of oxidative stress and corresponding antioxidant therapy in preclinical ALS by mining published quantitative experimental data and performing metadata analysis of 41 studies. In vitro aggregate analysis of innate oxidative stress inducers, glutamate and hydrogen peroxide, revealed 70-90% of cell death coincides to inducer exposure equivalent to 30-50% peak concentration (p < 0.05). A correlative plateau in cell death suggests oxidative stress impact is greatest in early-stage neurodegeneration. In vivo SOD1-G93A transgenic ALS mouse aggregate analysis of heat shock proteins (HSPs) revealed HSP levels are 30% lower in muscle than spine (p < 0.1). Overall spine HSP levels, including HSP70, are mildly upregulated in SOD1-G93A mice compared to wild type, but not significantly (p > 0.05). Thus, innate HSP compensatory responses to oxidative stress are simply insufficient, a result supportive of homeostatic system instability as central to ALS etiology. In vivo aggregate analysis of antioxidant therapy finds SOD1-G93A ALS mouse survival duration significantly increases by 11.2% (p << 0.001) but insignificantly decreases onset age by 2%. Thus, the aggregate antioxidant treatment effect on survival in preclinical ALS is not sufficient to overcome clinical heterogeneity, which explains the literature disparity between preclinical and clinical antioxidant survival benefit. The aggregate effect sizes on preclinical ALS survival and onset illustrate that present antioxidants, alone, are not sufficient to halt ALS, which underscores its multi-factorial nature. Nonetheless, antioxidant-treated SOD1-G93A ALS mice have significantly increased motor performance (p < 0.05) measured via rotarod. With a colossal aggregate preclinical effect size average of 59.6%, antioxidants are promising for increasing function/quality of life in clinical ALS patients, a premise worth exploration via low-risk nutritional supplements. Finally, more direct, quantitative measures of oxidative stress, antioxidant levels and bioavailability are key to developing powerful antioxidant therapeutics that can assert measurable impacts on redox homeostasis in the brain and spinal cord.}, }
@article {pmid29425965, year = {2018}, author = {Zhu, Z and Reiser, G}, title = {The small heat shock proteins, especially HspB4 and HspB5 are promising protectants in neurodegenerative diseases.}, journal = {Neurochemistry international}, volume = {115}, number = {}, pages = {69-79}, doi = {10.1016/j.neuint.2018.02.006}, pmid = {29425965}, issn = {1872-9754}, mesh = {Animals ; Brain/metabolism ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins/*drug effects/metabolism ; Heat-Shock Proteins, Small/metabolism ; Mice ; Neurodegenerative Diseases/*physiopathology ; Peptides/pharmacology ; alpha-Crystallin B Chain/*genetics ; }, abstract = {Small heat shock proteins (sHsps) are a group of proteins with molecular mass between 12 and 43 kDa. Currently, 11 members of this family have been classified, namely HspB1 to HspB11. HspB1, HspB2, HspB5, HspB6, HspB7, and HspB8, which are expressed in brain have been observed to be related to the pathology of neurodegenerative diseases, including Parkinson's, Alzheimer's, Alexander's disease, multiple sclerosis, and human immunodeficiency virus-associated dementia. Specifically, sHsps interact with misfolding and damaging protein aggregates, like Glial fibrillary acidic protein in AxD, β-amyloid peptides aggregates in Alzheimer's disease, Superoxide dismutase 1 in Amyotrophic lateral sclerosis and cytosine-adenine-guanine/polyglutamine (CAG/PolyQ) in Huntington's disease, Spinocerebellar ataxia type 3, Spinal-bulbar muscular atrophy, to reduce the toxicity or increase the clearance of these protein aggregates. The degree of HspB4 expression in brain is still debated. For neuroprotective mechanisms, sHsps attenuate mitochondrial dysfunctions, reduce accumulation of misfolded proteins, block oxidative/nitrosative stress, and minimize neuronal apoptosis and neuroinflammation, which are molecular mechanisms commonly accepted to mirror the progression and development of neurodegenerative diseases. The increasing incidence of the neurodegenerative diseases enhanced search for effective approaches to rescue neural tissue from degeneration with minimal side effects. sHsps have been found to exert neuroprotective functions. HspB5 has been emphasized to reduce the paralysis in a mouse model of experimental autoimmune encephalomyelitis, providing a therapeutic basis for the disease. In this review, we discuss the current understanding of the properties and the mechanisms of protection orchestrated by sHsps in the nervous system, highlighting the promising therapeutic role of sHsps in neurodegenerative diseases.}, }
@article {pmid29426953, year = {2018}, author = {Kahl, A and Blanco, I and Jackman, K and Baskar, J and Milaganur Mohan, H and Rodney-Sandy, R and Zhang, S and Iadecola, C and Hochrainer, K}, title = {Cerebral ischemia induces the aggregation of proteins linked to neurodegenerative diseases.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {2701}, pmid = {29426953}, issn = {2045-2322}, support = {R01 NS034179/NS/NINDS NIH HHS/United States ; S10 OD017992/OD/NIH HHS/United States ; S10 RR025449/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Brain Ischemia/*metabolism/*physiopathology ; Cerebral Infarction ; Cerebrovascular Circulation/physiology ; Hippocampus/metabolism ; Infarction, Middle Cerebral Artery/metabolism/physiopathology ; Ischemic Attack, Transient ; Male ; Mice ; Mice, Inbred C57BL ; Middle Cerebral Artery/physiopathology ; Neurodegenerative Diseases/metabolism ; Neurons/metabolism ; Protein Aggregates/physiology ; Proteomics/methods ; Reperfusion ; Reperfusion Injury/metabolism ; Small Ubiquitin-Related Modifier Proteins/metabolism ; Stroke/metabolism ; Tandem Mass Spectrometry ; Ubiquitin/metabolism ; }, abstract = {Protein aggregation critically affects cell viability in neurodegenerative diseases, but whether this also occurs in ischemic brain injury remains elusive. Prior studies report the post-ischemic aggregation of ubiquitin, small ubiquitin-related modifier (SUMO) and ribosomes, however whether other proteins are also affected is unknown. Here we employed a proteomic approach to identify the insoluble, aggregated proteome after cerebral ischemia. Mice underwent transient middle cerebral artery occlusion or sham-surgery. After 1-hour reperfusion, prior to apparent brain injury, mice were sacrificed and detergent-insoluble proteins were obtained and identified by nanoLC-MS/MS. Naturally existing insoluble proteins were determined in sham controls and aggregated proteins after cerebral ischemia/reperfusion were identified. Selected aggregated proteins found by proteomics were biochemically verified and aggregation propensities were studied during ischemia with or without reperfusion. We found that ischemia/reperfusion induces the aggregation of RNA-binding and heat-shock proteins, ubiquitin, SUMO and other proteins involved in cell signalling. RNA-binding proteins constitute the largest group of aggregating proteins in ischemia. These include TDP43, FUS, hnRNPA1, PSF/SFPQ and p54/NONO, all of which have been linked to neurodegeneration associated with amyotrophic lateral sclerosis and frontotemporal dementia. The aggregation of neurodegeneration-related disease proteins in cerebral ischemia unveils a previously unappreciated molecular overlap between neurodegenerative diseases and ischemic stroke.}, }
@article {pmid29515358, year = {2018}, author = {Seminary, ER and Sison, SL and Ebert, AD}, title = {Modeling Protein Aggregation and the Heat Shock Response in ALS iPSC-Derived Motor Neurons.}, journal = {Frontiers in neuroscience}, volume = {12}, number = {}, pages = {86}, pmid = {29515358}, issn = {1662-4548}, abstract = {Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder caused by the selective loss of the upper and lower motor neurons. Only 10% of all cases are caused by a mutation in one of the two dozen different identified genes, while the remaining 90% are likely caused by a combination of as yet unidentified genetic and environmental factors. Mutations in C9orf72, SOD1, or TDP-43 are the most common causes of familial ALS, together responsible for at least 60% of these cases. Remarkably, despite the large degree of heterogeneity, all cases of ALS have protein aggregates in the brain and spinal cord that are immunopositive for SOD1, TDP-43, OPTN, and/or p62. These inclusions are normally prevented and cleared by heat shock proteins (Hsps), suggesting that ALS motor neurons have an impaired ability to induce the heat shock response (HSR). Accordingly, there is evidence of decreased induction of Hsps in ALS mouse models and in human post-mortem samples compared to unaffected controls. However, the role of Hsps in protein accumulation in human motor neurons has not been fully elucidated. Here, we generated motor neuron cultures from human induced pluripotent stem cell (iPSC) lines carrying mutations in SOD1, TDP-43, or C9orf72. In this study, we provide evidence that despite a lack of overt motor neuron loss, there is an accumulation of insoluble, aggregation-prone proteins in iPSC-derived motor neuron cultures but that content and levels vary with genetic background. Additionally, although iPSC-derived motor neurons are generally capable of inducing the HSR when exposed to a heat stress, protein aggregation itself is not sufficient to induce the HSR or stress granule formation. We therefore conclude that ALS iPSC-derived motor neurons recapitulate key early pathological features of the disease and fail to endogenously upregulate the HSR in response to increased protein burden.}, }
@article {pmid29697388, year = {2018}, author = {Kvirkvelia, N and Shakarishvili, R and Kanashvili, T}, title = {[TRANSFORMATION OF MYASTHENIA GRAVIS INTO AMYOTROPHIC LATERAL SCLEROSIS, OR THEIR CONCOMITANCE? (CASE REVIEW)].}, journal = {Georgian medical news}, volume = {}, number = {276}, pages = {86-92}, pmid = {29697388}, issn = {1512-0112}, mesh = {Aged ; Amyotrophic Lateral Sclerosis/complications/*diagnosis/drug therapy/physiopathology ; Humans ; Male ; Myasthenia Gravis/complications/*diagnosis/drug therapy/physiopathology ; }, abstract = {The authors present a case of 75-year-old male patient with typical clinical and electroneuromyographic signs of Amyotrophic Lateral Sclerosis (ALS), manifested in 4 years after a diagnosis of generalized Myasthenia Gravis (MG) had been made. The aim of the article is to assess the possibility of pathogenetic integrated comorbidity of MG and ALS, which may have resulted from a common aberrant immune process and to emphasize the importance of detailed clinical analysis and adequate diagnostic methods essential for correct diagnosis and treatment. Only several cases of coexistence of MG and ALS have been described in medical literature. Exploring the pathogenetic association between MG and ALS may lead to dysregulation of thea immune system. Deficiency of T-regulatory cells, increased activity of atrophy-related atrogenes, anomalies of neuronal nitric oxide synthase can be found in both diseases. Immunoglobulin isolated from ALS patients can affect neuromuscular junction and activate AChRs, which plays an important role in the innervation and re-innervation of muscle fibers. Immunoglobulin also changes the function of Ca2+ channels. Blood level of circulatory Heat Shock Protein 70 (HSP70) antibodies in MG patients is elevated. HSP70 maintains normal conformation of cell proteins. Conversely, HSP70 antibodies cause HSP70's dysfunction and therefore, abnormal protein synthesis, which can be the main reason of neurodegenerative diseases, such as ALS. Experimental evidence indicates, that muscle and neuromuscular junctions may be initial targets of ALS. According to the "dying-back" hypothesis, neuromuscular junction damage and failure in MG patients may precede lower and upper motor neuron loss, and thus increase risk of developing ALS. Pathogenetic mechanisms of MG and ALS are the subjects of further studies. Refining the etiology of these two diseases will answer the question whether it is a transformation or a coexistence of MG and ALS in our case. The presented case demonstrates, that in spite of meeting all diagnostic criteria it is, sometimes, impossible to make the correct diagnosis. Only a detailed clinical analysis and adequate diagnostic methods contribute to correct diagnosis and adequate therapy.}, }
@article {pmid29973287, year = {2018}, author = {Mordes, DA and Prudencio, M and Goodman, LD and Klim, JR and Moccia, R and Limone, F and Pietilainen, O and Chowdhary, K and Dickson, DW and Rademakers, R and Bonini, NM and Petrucelli, L and Eggan, K}, title = {Dipeptide repeat proteins activate a heat shock response found in C9ORF72-ALS/FTLD patients.}, journal = {Acta neuropathologica communications}, volume = {6}, number = {1}, pages = {55}, pmid = {29973287}, issn = {2051-5960}, support = {R01 NS089742/NS/NINDS NIH HHS/United States ; R35 NS097273/NS/NINDS NIH HHS/United States ; P50 AG016574/AG/NIA NIH HHS/United States ; P30 AG062421/AG/NIA NIH HHS/United States ; P01 NS084974/NS/NINDS NIH HHS/United States ; R21 NS084528/NS/NINDS NIH HHS/United States ; R01 ES020395/ES/NIEHS NIH HHS/United States ; R35 NS097275/NS/NINDS NIH HHS/United States ; R01 NS077402/NS/NINDS NIH HHS/United States ; R01 NS063964/NS/NINDS NIH HHS/United States ; P50 AG005134/AG/NIA NIH HHS/United States ; T32 CA009216/CA/NCI NIH HHS/United States ; R01 NS088689/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Brain/*metabolism/pathology ; C9orf72 Protein/*genetics ; Cohort Studies ; DNA Repeat Expansion/*genetics ; Dipeptides ; Disease Models, Animal ; Drosophila ; Eye/pathology ; Female ; Frontotemporal Lobar Degeneration/*genetics/pathology ; Gene Expression Regulation/*genetics ; Glial Fibrillary Acidic Protein/metabolism ; Heat Shock Transcription Factors/genetics/metabolism ; Heat-Shock Response/*physiology ; Humans ; Male ; Neurons/metabolism ; Signal Transduction/physiology ; Stem Cells/metabolism ; }, abstract = {A hexanucleotide (GGGGCC) repeat expansion in C9ORF72 is the most common genetic contributor to amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Reduced expression of the C9ORF72 gene product has been proposed as a potential contributor to disease pathogenesis. Additionally, repetitive RNAs and dipeptide repeat proteins (DPRs), such as poly-GR, can be produced by this hexanucleotide expansion that disrupt a number of cellular processes, potentially contributing to neural degeneration. To better discern which of these mechanisms leads to disease-associated changes in patient brains, we analyzed gene expression data generated from the cortex and cerebellum. We found that transcripts encoding heat shock proteins (HSPs) regulated by the HSF1 transcription factor were significantly induced in C9ORF72-ALS/FTLD patients relative to both sporadic ALS/FTLD cases and controls. Treatment of human neurons with chemically synthesized DPRs was sufficient to activate a similar transcriptional response. Expression of GGGGCC repeats and also poly-GR in the brains of Drosophila lead to the upregulation of HSF1 and the same highly-conserved HSPs. Additionally, HSF1 was a modifier of poly-GR toxicity in Drosophila. Our results suggest that the expression of DPRs are associated with upregulation of HSF1 and activation of a heat shock response in C9ORF72-ALS/FTLD.}, }
@article {pmid30099725, year = {2018}, author = {Kitamura, A and Iwasaki, N and Kinjo, M}, title = {Molecular chaperone HSP70 prevents formation of inclusion bodies of the 25-kDa C-terminal fragment of TDP-43 by preventing aggregate accumulation.}, journal = {Cell stress &amp; chaperones}, volume = {23}, number = {6}, pages = {1177-1183}, pmid = {30099725}, issn = {1466-1268}, mesh = {Amyotrophic Lateral Sclerosis/*pathology ; Animals ; Cell Line, Tumor ; Cytoplasm/metabolism ; DNA-Binding Proteins/*metabolism ; HSP70 Heat-Shock Proteins/*physiology ; Humans ; Inclusion Bodies/*metabolism ; Mice ; Peptide Fragments/*metabolism ; Protein Aggregation, Pathological ; }, abstract = {Transactive response DNA/RNA-binding protein 43-kDa (TDP-43) C-terminal fragments, such as a 25-kDa fragment (TDP-25), have been identified as a ubiquitinated and phosphorylated components of inclusion bodies (IBs) in motor neurons from amyotrophic lateral sclerosis patients. Cells contain proteins that function as molecular chaperones and prevent aggregate formation of misfolded and aggregation-prone proteins. Recently, we reported that heat shock protein (HSP)70, an abundant molecular chaperone, binds to TDP-25 in an ATP-dependent manner; however, whether HSP70 can prevent the formation of TDP-25-related IBs remains unknown. Here, we showed that HSP70 prevented TDP-25 aggregation according to green fluorescent protein-tagged TDP-25 (G-TDP-25) colocalization in the cytoplasm with mCherry-tagged HSP70 (HSP70-R). The mobile fraction of HSP70-R in the cytoplasmic IBs associated with G-TDP-25 increased relative to that of G-TDP-25, suggesting that HSP70 strongly bound to G-TDP-25 in the IBs, whereas a portion remained dissociated from the IBs. Importantly, the proportion of G-TDP-25 IBs was significantly decreased by HSP70-R overexpression; however, G-TDP-25 levels in the insoluble fraction remained unchanged by HSP70-R overexpression, suggesting that G-TDP-25 formed aggregated species that cannot be dissolved, even in the presence of strong detergents. These results indicated that HSP70 prevented the accumulation of G-TDP-25 aggregates in cytoplasmic IBs, but was insufficient for G-TDP-25 disassembly and solubilization.}, }
@article {pmid30335591, year = {2019}, author = {Rusmini, P and Cortese, K and Crippa, V and Cristofani, R and Cicardi, ME and Ferrari, V and Vezzoli, G and Tedesco, B and Meroni, M and Messi, E and Piccolella, M and Galbiati, M and Garrè, M and Morelli, E and Vaccari, T and Poletti, A}, title = {Trehalose induces autophagy via lysosomal-mediated TFEB activation in models of motoneuron degeneration.}, journal = {Autophagy}, volume = {15}, number = {4}, pages = {631-651}, pmid = {30335591}, issn = {1554-8635}, mesh = {Amyotrophic Lateral Sclerosis/drug therapy/metabolism ; Animals ; Autophagosomes/drug effects/enzymology/metabolism ; Autophagy/*drug effects/genetics ; Autophagy-Related Proteins/genetics/metabolism ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/chemistry/genetics/*metabolism ; Bulbo-Spinal Atrophy, X-Linked/drug therapy/metabolism ; Calcineurin/genetics/*metabolism ; Calcium/metabolism ; Cell Differentiation ; Cell Nucleus/metabolism ; Down-Regulation/genetics ; Humans ; Induced Pluripotent Stem Cells/enzymology/metabolism/pathology/ultrastructure ; Lysosomes/drug effects/enzymology/*metabolism/ultrastructure ; Membrane Proteins/genetics/metabolism ; Mice ; Microtubule-Associated Proteins/metabolism ; Motor Neurons/enzymology/*metabolism/*pathology/ultrastructure ; Neuroprotection/drug effects/genetics ; RNA, Small Interfering/genetics/metabolism ; Sequestosome-1 Protein/metabolism ; Signal Transduction/drug effects/genetics ; Trehalose/analogs &amp; derivatives/*pharmacology ; Tripeptidyl-Peptidase 1 ; Unfolded Protein Response/genetics ; }, abstract = {Macroautophagy/autophagy, a defense mechanism against aberrant stresses, in neurons counteracts aggregate-prone misfolded protein toxicity. Autophagy induction might be beneficial in neurodegenerative diseases (NDs). The natural compound trehalose promotes autophagy via TFEB (transcription factor EB), ameliorating disease phenotype in multiple ND models, but its mechanism is still obscure. We demonstrated that trehalose regulates autophagy by inducing rapid and transient lysosomal enlargement and membrane permeabilization (LMP). This effect correlated with the calcium-dependent phosphatase PPP3/calcineurin activation, TFEB dephosphorylation and nuclear translocation. Trehalose upregulated genes for the TFEB target and regulator Ppargc1a, lysosomal hydrolases and membrane proteins (Ctsb, Gla, Lamp2a, Mcoln1, Tpp1) and several autophagy-related components (Becn1, Atg10, Atg12, Sqstm1/p62, Map1lc3b, Hspb8 and Bag3) mostly in a PPP3- and TFEB-dependent manner. TFEB silencing counteracted the trehalose pro-degradative activity on misfolded protein causative of motoneuron diseases. Similar effects were exerted by trehalase-resistant trehalose analogs, melibiose and lactulose. Thus, limited lysosomal damage might induce autophagy, perhaps as a compensatory mechanism, a process that is beneficial to counteract neurodegeneration. Abbreviations: ALS: amyotrophic lateral sclerosis; AR: androgen receptor; ATG: autophagy related; AV: autophagic vacuole; BAG3: BCL2-associated athanogene 3; BECN1: beclin 1, autophagy related; CASA: chaperone-assisted selective autophagy; CTSB: cathepsin b; DAPI: 4',6-diamidino-2-phenylindole; DMEM: Dulbecco's modified Eagle's medium; EGFP: enhanced green fluorescent protein; fALS, familial amyotrophic lateral sclerosis; FRA: filter retardation assay; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GLA: galactosidase, alpha; HD: Huntington disease; hIPSCs: human induced pluripotent stem cells; HSPA8: heat shock protein A8; HSPB8: heat shock protein B8; IF: immunofluorescence analysis; LAMP1: lysosomal-associated membrane protein 1; LAMP2A: lysosomal-associated membrane protein 2A; LGALS3: lectin, galactose binding, soluble 3; LLOMe: L-leucyl-L-leucine methyl ester; LMP: lysosomal membrane permeabilization; Lys: lysosomes; MAP1LC3B: microtubule-associated protein 1 light chain 3 beta; MCOLN1: mucolipin 1; mRNA: messenger RNA; MTOR: mechanistic target of rapamycin kinase; NDs: neurodegenerative diseases; NSC34: neuroblastoma x spinal cord 34; PBS: phosphate-buffered saline; PD: Parkinson disease; polyQ: polyglutamine; PPARGC1A: peroxisome proliferative activated receptor, gamma, coactivator 1 alpha; PPP3CB: protein phosphatase 3, catalytic subunit, beta isoform; RT-qPCR: real-time quantitative polymerase chain reaction; SBMA: spinal and bulbar muscular atrophy; SCAs: spinocerebellar ataxias; siRNA: small interfering RNA; SLC2A8: solute carrier family 2, (facilitated glucose transporter), member 8; smNPCs: small molecules neural progenitors cells; SOD1: superoxide dismutase 1; SQSTM1/p62: sequestosome 1; STED: stimulated emission depletion; STUB1: STIP1 homology and U-box containing protein 1; TARDBP/TDP-43: TAR DNA binding protein; TFEB: transcription factor EB; TPP1: tripeptidyl peptidase I; TREH: trehalase (brush-border membrane glycoprotein); WB: western blotting; ZKSCAN3: zinc finger with KRAB and SCAN domains 3.}, }
@article {pmid30346063, year = {2019}, author = {Gorter, RP and Stephenson, J and Nutma, E and Anink, J and de Jonge, JC and Baron, W and Jahreiβ, MC and Belien, JAM and van Noort, JM and Mijnsbergen, C and Aronica, E and Amor, S}, title = {Rapidly progressive amyotrophic lateral sclerosis is associated with microglial reactivity and small heat shock protein expression in reactive astrocytes.}, journal = {Neuropathology and applied neurobiology}, volume = {45}, number = {5}, pages = {459-475}, pmid = {30346063}, issn = {1365-2990}, mesh = {Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/metabolism/*pathology ; Astrocytes/*metabolism ; Disease Progression ; Female ; Heat-Shock Proteins/*metabolism ; Humans ; Male ; Microglia/*metabolism ; Middle Aged ; Motor Neurons/metabolism/pathology ; Spinal Cord/metabolism/pathology ; }, abstract = {AIMS: Amyotrophic lateral sclerosis (ALS) is a chronic neurodegenerative disease characterized by progressive loss of motor neurons, muscle weakness, spasticity, paralysis and death usually within 2-5 years of onset. Neuroinflammation is a hallmark of ALS pathology characterized by activation of glial cells, which respond by upregulating small heat shock proteins (HSPBs), but the exact underlying pathological mechanisms are still largely unknown. Here, we investigated the association between ALS disease duration, lower motor neuron loss, TARDNA-binding protein 43 (TDP-43) pathology, neuroinflammation and HSPB expression.<br><br>METHODS: With immunohistochemistry, we examined HSPB1, HSPB5, HSPB6, HSPB8 and HSP16.2 expression in cervical, thoracic and sacral spinal cord regions in 12 ALS cases, seven with short disease duration (SDD), five with moderate disease duration (MDD), and ten age-matched controls. Expression was quantified using ImageJ to examine HSP expression, motor neuron numbers, microglial and astrocyte density and phosphorylated TDP-43 (pTDP-43+) inclusions.<br><br>RESULTS: SDD was associated with elevated HSPB5 and 8 expression in lateral tract astrocytes, while HSP16.2 expression was increased in astrocytes in MDD cases. SDD cases had higher numbers of motor neurons and microglial activation than MDD cases, but similar levels of motor neurons with pTDP-43+ inclusions.<br><br>CONCLUSIONS: Increased expression of several HSPBs in lateral column astrocytes suggests that astrocytes play a role in the pathogenesis of ALS. SDD is associated with increased microgliosis, HSPB5 and 8 expression in astrocytes, and only minor changes in motor neuron loss. This suggests that the interaction between motor neurons, microglia and astrocytes determines neuronal fate and functional decline in ALS.}, }
@article {pmid30454072, year = {2018}, author = {Shaw, MP and Higginbottom, A and McGown, A and Castelli, LM and James, E and Hautbergue, GM and Shaw, PJ and Ramesh, TM}, title = {Stable transgenic C9orf72 zebrafish model key aspects of the ALS/FTD phenotype and reveal novel pathological features.}, journal = {Acta neuropathologica communications}, volume = {6}, number = {1}, pages = {125}, pmid = {30454072}, issn = {2051-5960}, support = {RAMESH/APR17/854-791/MNDA_/Motor Neurone Disease Association/United Kingdom ; RAMESH/OCT14/875-792/MNDA_/Motor Neurone Disease Association/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/*genetics/*pathology ; Animals ; Animals, Genetically Modified ; C9orf72 Protein/*genetics/metabolism ; Cell Line ; Cells, Cultured ; Disease Models, Animal ; Embryo, Nonmammalian ; Frontotemporal Dementia/*genetics/*pathology ; Gene Expression Regulation/genetics ; Green Fluorescent Proteins/genetics/metabolism ; Heat-Shock Response ; Humans ; Locomotion/genetics ; Mice ; Motor Neurons/pathology ; Muscles/metabolism/pathology/ultrastructure ; Superoxide Dismutase-1/metabolism ; Transfection ; Zebrafish ; }, abstract = {A hexanucleotide repeat expansion (HRE) within the chromosome 9 open reading frame 72 (C9orf72) gene is the most prevalent cause of amyotrophic lateral sclerosis/fronto-temporal dementia (ALS/FTD). Current evidence suggests HREs induce neurodegeneration through accumulation of RNA foci and/or dipeptide repeat proteins (DPR). C9orf72 patients are known to have transactive response DNA binding protein 43 kDa (TDP-43) proteinopathy, but whether there is further cross over between C9orf72 pathology and the pathology of other ALS sub-types has yet to be revealed.To address this, we generated and characterised two zebrafish lines expressing C9orf72 HREs. We also characterised pathology in human C9orf72-ALS cases. In addition, we utilised a reporter construct that expresses DsRed under the control of a heat shock promoter, to screen for potential therapeutic compounds.Both zebrafish lines showed accumulation of RNA foci and DPR. Our C9-ALS/FTD zebrafish model is the first to recapitulate the motor deficits, cognitive impairment, muscle atrophy, motor neuron loss and mortality in early adulthood observed in human C9orf72-ALS/FTD. Furthermore, we identified that in zebrafish, human cell lines and human post-mortem tissue, C9orf72 expansions activate the heat shock response (HSR). Additionally, HSR activation correlated with disease progression in our C9-ALS/FTD zebrafish model. Lastly, we identified that the compound ivermectin, as well as riluzole, reduced HSR activation in both C9-ALS/FTD and SOD1 zebrafish models.Thus, our C9-ALS/FTD zebrafish model is a stable transgenic model which recapitulates key features of human C9orf72-ALS/FTD, and represents a powerful drug-discovery tool.}, }
@article {pmid30618614, year = {2018}, author = {Lissouba, A and Liao, M and Kabashi, E and Drapeau, P}, title = {Transcriptomic Analysis of Zebrafish TDP-43 Transgenic Lines.}, journal = {Frontiers in molecular neuroscience}, volume = {11}, number = {}, pages = {463}, pmid = {30618614}, issn = {1662-5099}, abstract = {Amyotrophic lateral sclerosis (ALS) is a late-onset progressive neurodegenerative disorder that affects both upper and lower motor neurons, leading to muscle atrophy with spasticity and eventual death in 3-5 years after the disease onset. More than 50 mutations linked to ALS have been found in the gene TARDBP, encoding the protein TDP-43 that is the predominant component of neuronal inclusions in ALS. TDP-43 is an RNA binding protein with glycine-rich domains that binds to more than 6,000 RNAs in the human brain. However, ALS-related mutations do not appear to affect the function of these genes, indicating that a toxic gain-of-function may occur. We generated transgenic zebrafish lines expressing human TDP-43, either the wild-type form or the ALS-causative G348C mutation identified in a subset of ALS patients, with the transgene expression driven by an inducible heat shock promoter in order to bypass a potential early mortality. The expression of the mutant but not the wild-type human TDP-43 in zebrafish embryos induced a reduction of the locomotor activity in response to touch compared to controls and moderate axonopathy of the motor neurons of the spinal cord, with premature branching of the main axonal branch, recapitulating previous results obtained by mRNA injections. We used these lines to investigate transcriptomic changes due to the presence of mutant TDP-43 using RNA sequencing and have found 159 genes that are differentially expressed compared to control, with 67 genes up-regulated and 92 genes down-regulated. These transcriptomic changes are in line with recent transcriptomic data obtained in mouse models, indicating that these zebrafish transgenic lines are adequate to further study TDP-43-related ALS.}, }
@article {pmid30635082, year = {2019}, author = {Mesika, R and Reichmann, D}, title = {When safeguarding goes wrong: Impact of oxidative stress on protein homeostasis in health and neurodegenerative disorders.}, journal = {Advances in protein chemistry and structural biology}, volume = {114}, number = {}, pages = {221-264}, doi = {10.1016/bs.apcsb.2018.11.001}, pmid = {30635082}, issn = {1876-1631}, mesh = {Animals ; *Health ; Heat Shock Transcription Factors/*metabolism ; Heat-Shock Response ; *Homeostasis ; Humans ; NF-E2-Related Factor 2/metabolism ; Neurodegenerative Diseases/*metabolism ; *Oxidative Stress ; Reactive Oxygen Species/metabolism ; }, abstract = {Cellular redox status is an established player in many different cellular functions. The buildup of oxidants within the cell is tightly regulated to maintain a balance between the positive and negative outcomes of cellular oxidants. Proteins are highly sensitive to oxidation, since modification can cause widespread unfolding and the formation of toxic aggregates. In response, cells have developed highly regulated systems that contribute to the maintenance of both the global redox status and protein homeostasis at large. Changes to these systems have been found to correlate with aging and age-related disorders, such as neurodegenerative pathologies. This raises intriguing questions as to the source of the imbalance in the redox and protein homeostasis systems, their interconnectivity, and their role in disease progression. Here we focus on the crosstalk between the redox and protein homeostasis systems in neurodegenerative diseases, specifically in Alzheimer's, Parkinson's, and ALS. We elaborate on some of the main players of the stress response systems, including the master regulators of oxidative stress and the heat shock response, Nrf2 and Hsf1, which are essential features of protein folding, and mediators of protein turnover. We illustrate the elegant mechanisms used by these components to provide an immediate response, including protein plasticity controlled by redox-sensing cysteines and the recruitment of naive proteins to the redox homeostasis array that act as chaperons in an ATP-independent manner.}, }
@article {pmid30654731, year = {2019}, author = {Arhzaouy, K and Papadopoulos, C and Schulze, N and Pittman, SK and Meyer, H and Weihl, CC}, title = {VCP maintains lysosomal homeostasis and TFEB activity in differentiated skeletal muscle.}, journal = {Autophagy}, volume = {15}, number = {6}, pages = {1082-1099}, pmid = {30654731}, issn = {1554-8635}, support = {K02 AG042095/AG/NIA NIH HHS/United States ; T32 NS007205/NS/NINDS NIH HHS/United States ; R01 AG031867/AG/NIA NIH HHS/United States ; K24 AR073317/AR/NIAMS NIH HHS/United States ; R01 AR068797/AR/NIAMS NIH HHS/United States ; }, mesh = {Animals ; Autophagy/drug effects/*genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics/*metabolism ; Endoplasmic Reticulum Chaperone BiP ; HeLa Cells ; Homeostasis/genetics ; Humans ; *Lysosomes/drug effects/genetics/pathology/ultrastructure ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle Fibers, Skeletal/cytology/*metabolism/ultrastructure ; Valosin Containing Protein/antagonists &amp; inhibitors/genetics/*metabolism ; }, abstract = {Differentiated tissue is particularly vulnerable to alterations in protein and organelle homeostasis. The essential protein VCP, mutated in hereditary inclusion body myopathy, amyotrophic lateral sclerosis and frontotemporal dementia, is critical for efficient clearance of misfolded proteins and damaged organelles in dividing cells, but its role in terminally differentiated tissue affected by disease mutations is less clear. To understand the relevance of VCP in differentiated tissue, we inactivated it in skeletal muscle of adult mice. Surprisingly, knockout muscle demonstrated a necrotic myopathy with increased macroautophagic/autophagic proteins and damaged lysosomes. This was not solely due to a defect in autophagic degradation because age-matched mice with muscle inactivation of the autophagy essential protein, ATG5, did not demonstrate a myopathy. Notably, myofiber necrosis was preceded by upregulation of LGALS3/Galectin-3, a marker of damaged lysosomes, and TFEB activation, suggesting early defects in the lysosomal system. Consistent with that, myofiber necrosis was recapitulated by chemical induction of lysosomal membrane permeabilization (LMP) in skeletal muscle. Moreover, TFEB was activated after LMP in cells, but activation and nuclear localization of TFEB persisted upon VCP inactivation or disease mutant expression. Our data identifies VCP as central mediator of both lysosomal clearance and biogenesis in skeletal muscle. Abbreviations: AAA: ATPases Associated with diverse cellular Activities; TUBA1A/α-tubulin: tubulin alpha 1a; ATG5: autophagy related 5; ATG7: autophagy related 7; ACTA1: actin alpha 1, skeletal muscle; CLEAR: coordinated lysosomal expression and regulation; CTSB/D: cathepsin B/D; Ctrl: control; DAPI: diamidino-2-phenylindole; EBSS: Earle's balanced salt solution; ELDR: endolysosomal damage response; ESCRT: endosomal sorting complexes required for transport; Gastroc/G: gastrocnemius; H&amp;E: hematoxylin and eosin; HSPA5/GRP78: heat shock protein family A (Hsp70) member 5; IBMPFD/ALS: inclusion body myopathy associated with Paget disease of the bone, frontotemporal dementia and amyotrophic lateral sclerosis; i.p.: intraperitoneal; LAMP1/2: lysosomal-associated membrane protein 1/2; LLOMe: Leu-Leu methyl ester hydrobromide; LGALS3/Gal3: galectin 3; LMP: lysosomal membrane permeabilization; MTOR: mechanistic target of rapamycin kinase; MYL1: myosin light chain 1; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MSP: multisystem proteinopathy; PBS: phosphate-buffered saline; PCR: polymerase chain reaction; Quad/Q: quadriceps; RHEB: Ras homolog, mTORC1 binding; SQSTM1: sequestosome 1; TFEB: transcription factor EB; TA: tibialis anterior; siRNA: small interfering RNA; SQSTM1/p62, sequestosome 1; TARDBP/TDP-43: TAR DNA binding protein; TBS: Tris-buffered saline; TXFN, tamoxifen; UBXN6/UBXD1: UBX domain protein 6; VCP: valosin containing protein; WT: wild-type.}, }
@article {pmid30669930, year = {2019}, author = {Haidar, M and Asselbergh, B and Adriaenssens, E and De Winter, V and Timmermans, JP and Auer-Grumbach, M and Juneja, M and Timmerman, V}, title = {Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies.}, journal = {Autophagy}, volume = {15}, number = {6}, pages = {1051-1068}, pmid = {30669930}, issn = {1554-8635}, support = {P 27634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Amyotrophic Lateral Sclerosis/genetics ; Autophagosomes/*metabolism/ultrastructure ; Autophagy/genetics ; Charcot-Marie-Tooth Disease/*genetics ; Chromatography, Liquid ; HeLa Cells ; Heat-Shock Proteins/chemistry/*genetics/metabolism ; Humans ; Induced Pluripotent Stem Cells/drug effects/metabolism ; Molecular Chaperones/chemistry/*genetics/metabolism ; Motor Neurons/*metabolism/pathology ; Muscular Atrophy, Spinal/*genetics ; Mutation ; Protein Domains ; Sequestosome-1 Protein/chemistry/genetics/*metabolism ; Tandem Mass Spectrometry ; }, abstract = {HSPB1 (heat shock protein family B [small] member 1) is a ubiquitously expressed molecular chaperone. Most mutations in HSPB1 cause axonal Charcot-Marie-Tooth neuropathy and/or distal hereditary motor neuropathy. In this study we show that mutations in HSPB1 lead to impairment of macroautophagic/autophagic flux. In HSPB1 knockout cells, we demonstrate that HSPB1 is necessary for autophagosome formation, which was rescued upon re-expression of HSPB1. Employing a label-free LC-MS/MS analysis on the various HSPB1 variants (wild type and mutants), we identified autophagy-specific interactors. We reveal that the wild-type HSPB1 protein binds to the autophagy receptor SQSTM1/p62 and that the PB1 domain of SQSTM1 is essential for this interaction. Mutations in HSPB1 lead to a decrease in the formation of SQSTM1/p62 bodies, and subsequent impairment of phagophore formation, suggesting a regulatory role for HSPB1 in autophagy via interaction with SQSTM1. Remarkably, autophagy deficits could also be confirmed in patient-derived motor neurons thereby indicating that the impairment of autophagy might be one of the pathomechanisms by which mutations in HSPB1 lead to peripheral neuropathy. Abbreviations: ACD: alpha-crystallin domain; ALS: amyotrophic lateral sclerosis; ATG14: autophagy related 14; BAG1/3: BCL2 associated athanogene 1/3; CMT: Charcot-Marie-Tooth; dHMN: distal hereditary motor neuropathy; GFP: green fluorescent protein; HSPA8: heat shock protein family A (Hsp70) member 8; HSPB1/6/8: heat shock protein family B (small) member 1/6/8; LIR: LC3-interacting region; LC3B: microtubule associated protein 1 light chain 3 beta; PB1: Phox and Bem1; SQSTM1: sequestosome 1; STUB1/CHIP: STIP1 homology and U-box containing protein 1; UBA: ubiquitin-associated; WIPI1: WD repeat domain, phosphoinositide interacting 1; WT: wild-type.}, }
@article {pmid31142143, year = {2019}, author = {Strayer, AL and Dennys-Rivers, CN and Ricart, KC and Bae, N and Beckman, JS and Franco, MC and Estevez, AG}, title = {Ligand-independent activation of the P2X7 receptor by Hsp90 inhibition stimulates motor neuron apoptosis.}, journal = {Experimental biology and medicine (Maywood, N.J.)}, volume = {244}, number = {11}, pages = {901-914}, pmid = {31142143}, issn = {1535-3699}, support = {R01 NS036761/NS/NINDS NIH HHS/United States ; R01 NS102479/NS/NINDS NIH HHS/United States ; }, mesh = {Adenosine Triphosphate/metabolism ; Animals ; Apoptosis/*drug effects ; Benzoquinones/*pharmacology ; Cells, Cultured ; Down-Regulation/drug effects ; Ethylenediamines/metabolism ; HSP90 Heat-Shock Proteins/*metabolism ; Lactams, Macrocyclic/*pharmacology ; Ligands ; Mice ; Motor Neurons/*drug effects/*metabolism ; Neurodegenerative Diseases/drug therapy/metabolism ; Oxidative Stress/drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X7/*metabolism ; Signal Transduction/drug effects ; }, abstract = {UNLABELLED: Activation of the extracellular ATP ionotropic receptor P2X7 stimulates motor neuron apoptosis, whereas its inhibition in cell and animal models of amyotrophic lateral sclerosis can be protective. These observations suggest that P2X7 receptor activation is relevant to motor neuron disease and that it could be targeted for therapeutic development. Heat shock protein 90 (Hsp90) is an integral regulatory component of the P2X7 receptor complex, antagonizing ligand-induced receptor activation. Here, we show that the repressive activity of Hsp90 on P2X7 receptor activation in primary motor neurons is highly sensitive to inhibition. Primary motor neurons in culture are 100-fold more sensitive to Hsp90 inhibition by geldanamycin than other neuronal populations. Pharmacological inhibition and down-regulation of the P2X7 receptor prevented motor neuron apoptosis triggered by Hsp90 inhibition, which occurred in the absence of extracellular ATP. These observations suggest that inhibition of a seemingly motor neuron specific pool of Hsp90 leads to ligand independent activation of P2X7 receptor and motor neuron death. Downstream of Hsp90 inhibition, P2X7 receptor activated the phosphatase and tensin homolog (TPEN), which in turn suppressed the pro-survival phosphatidyl inositol 3 kinase (PI3K)/Akt pathway, leading to Fas-dependent motor neuron apoptosis. Conditions altering the interaction between P2X7 receptor and Hsp90, such as recruitment of Hsp90 to other subcellular compartments under stress conditions, or nitration following oxidative stress can induce motor neuron death. These findings may have broad implications in neurodegenerative disorders, including amyotrophic lateral sclerosis, in which activation of P2X7 receptor may be involved in both autonomous and non-autonomous motor neurons death.<br><br>IMPACT STATEMENT: Here we show that a motor neuron specific pool of Hsp90 that is highly sensitive to geldanamycin inhibition represses ligand-independent activation of P2X7 receptor and is critical to motor neuron survival. Activation of P2X7 receptor by Hsp90 inhibition triggers motor neuron apoptosis through the activation of PTEN, which in turn inhibits the PI3 kinase/Akt survival pathway. Thus, inhibition of Hsp90 for therapeutic applications may have the unexpected negative consequence of decreasing the activity of trophic pathways in motor neurons. The inhibition of Hsp90 as a therapeutic approach may require the identification of the Hsp90 complexes involved in pathogenic processes and the development of inhibitors selective for these complexes.}, }
@article {pmid31152038, year = {2019}, author = {Mandrioli, J and Crippa, V and Cereda, C and Bonetto, V and Zucchi, E and Gessani, A and Ceroni, M and Chio, A and D'Amico, R and Monsurrò, MR and Riva, N and Sabatelli, M and Silani, V and Simone, IL and Sorarù, G and Provenzani, A and D'Agostino, VG and Carra, S and Poletti, A}, title = {Proteostasis and ALS: protocol for a phase II, randomised, double-blind, placebo-controlled, multicentre clinical trial for colchicine in ALS (Co-ALS).}, journal = {BMJ open}, volume = {9}, number = {5}, pages = {e028486}, pmid = {31152038}, issn = {2044-6055}, mesh = {Adult ; Aged ; Amyotrophic Lateral Sclerosis/*drug therapy/metabolism/physiopathology ; Autophagy/drug effects/*physiology ; Biomarkers ; Clinical Trials, Phase II as Topic ; Colchicine/pharmacokinetics/*therapeutic use ; DNA-Binding Proteins/antagonists &amp; inhibitors ; Disease Progression ; Double-Blind Method ; Female ; HSP20 Heat-Shock Proteins/*metabolism ; Heat-Shock Proteins ; Humans ; Male ; Middle Aged ; Molecular Chaperones ; Motor Neurons/drug effects/physiology ; Neuroprotective Agents/pharmacokinetics/*therapeutic use ; Proteostasis/*drug effects ; Randomized Controlled Trials as Topic ; Treatment Outcome ; Young Adult ; }, abstract = {INTRODUCTION: Disruptions of proteasome and autophagy systems are central events in amyotrophic lateral sclerosis (ALS) and support the urgent need to find therapeutic compounds targeting these processes. The heat shock protein B8 (HSPB8) recognises and promotes the autophagy-mediated removal of misfolded mutant SOD1 and TDP-43 fragments from ALS motor neurons (MNs), as well as aggregating species of dipeptides produced in C9ORF72-related diseases. In ALS-SOD1 mice and in human ALS autopsy specimens, HSPB8 is highly expressed in spinal cord MNs that survive at the end stage of disease. Moreover, the HSPB8-BAG3-HSP70 complex maintains granulostasis, which avoids conversion of dynamic stress granules (SGs) into aggregation-prone assemblies. We will perform a randomised clinical trial (RCT) with colchicine, which enhances the expression of HSPB8 and of several autophagy players, blocking TDP-43 accumulation and exerting crucial activities for MNs function.<br><br>METHODS AND ANALYSIS: Colchicine in amyotrophic lateral sclerosis (Co-ALS) is a double-blind, placebo-controlled, multicentre, phase II RCT. ALS patients will be enrolled in three groups (placebo, colchicine 0.01&#x2009;mg/day and colchicine 0.005&#x2009;mg/day) of 18 subjects treated with riluzole; treatment will last 30 weeks, and follow-up will last 24 weeks. The primary aim is to assess whether colchicine decreases disease progression as measured by ALS Functional Rating Scale - Revised (ALSFRS-R) at baseline and at treatment end. Secondary aims include assessment of (1) safety and tolerability of Colchicine in patiets with ALS; (2) changes in cellular activity (autophagy, protein aggregation, and SG and exosome secretion) and in biomarkers of disease progression (neurofilaments); (3) survival and respiratory function and (4) quality of life. Preclinical studies with a full assessment of autophagy and neuroinflammation biomarkers in fibroblasts, peripheral blood mononuclear cells and lymphoblasts will be conducted in parallel with clinic assessment to optimise time and resources.<br><br>ETHICS AND DISSEMINATION: The study protocol was approved by the Ethics Committee of Area Vasta Emilia Nord and by Agenzia Italiana del Farmaco (EUDRACT N.2017-004459-21) based on the Declaration of Helsinki. This research protocol was written without patient involvement. Patients' association will be involved in disseminating the study design and results. Results will be presented during scientific symposia or published in scientific journals.<br><br>TRIAL REGISTRATION NUMBER: EUDRACT 2017-004459-21; NCT03693781; Pre-results.}, }
@article {pmid31168740, year = {2019}, author = {Clarke, BE and Gil, RS and Yip, J and Kalmar, B and Greensmith, L}, title = {Regional differences in the inflammatory and heat shock response in glia: implications for ALS.}, journal = {Cell stress &amp; chaperones}, volume = {24}, number = {5}, pages = {857-870}, pmid = {31168740}, issn = {1466-1268}, support = {/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Mice ; *Amyotrophic Lateral Sclerosis/immunology/pathology ; *Brain/immunology/pathology ; Cells, Cultured ; *Heat-Shock Response/physiology ; HSP70 Heat-Shock Proteins/immunology ; *Inflammation/immunology ; Mice, Inbred C57BL ; *Neuroglia/immunology/pathology ; *Spinal Cord/immunology/pathology ; Primary Cell Culture ; Disease Models, Animal ; }, abstract = {Preferential neuronal vulnerability is characteristic of several neurodegenerative diseases including the motor neuron disease amyotrophic lateral sclerosis (ALS). It is well established that glia play a critical role in ALS, but it is unknown whether regional differences in the ability of glia to support motor neurons contribute to the specific pattern of neuronal degeneration. In this study, using primary mixed glial cultures from different mouse CNS regions (spinal cord and cortex), we examined whether regional differences exist in key glial pathways that contribute to, or protect against, motor neuron degeneration. Specifically, we examined the NF-κB-mediated inflammatory pathway and the cytoprotective heat shock response (HSR). Glial cultures were treated with pro-inflammatory stimuli, tumour necrosis factor-ɑ/lipopolysaccharide or heat stressed to stimulate the inflammatory and HSR respectively. We found that spinal cord glia expressed more iNOS and produced more NO compared to cortical glia in response to inflammatory stimuli. Intriguingly, we found that expression of ALS-causing SOD1[G93A] did not elevate the levels of NO in spinal cord glia. However, activation of the stress-responsive HSR was attenuated in SOD1[G93A] cultures, with a reduced Hsp70 induction in response to stressful stimuli. Exposure of spinal cord glia to heat shock in combination with inflammatory stimuli reduced the activation of the inflammatory response. The results of this study suggest that impaired heat shock response in SOD1[G93A] glia may contribute to the exacerbated inflammatory reactions observed in ALS mice. Graphical abstract Mixed primary glial cultures were established from cortical and spinal cord regions of wild-type mice and mice expressing ALS-causing mutant human SOD1 and the inflammatory and heat shock responses were investigated in these cultures. In the absence of stress, all cultures appeared to have similar cellular composition, levels of inflammatory mediators and similar expression level of heat shock proteins. When stimulated, spinal cord glia were more reactive and activated the inflammatory pathway more readily than cortical glia; this response was similar in wild-type and SOD1[G93A] glial cultures. Although the heat shock response was similar in spinal cord and cortical glial, in SOD1[G93A] expressing glia from both the spinal cord and cortex, the induction of heat shock response was diminished. This impaired heat shock response in SOD1[G93A] glia may therefore contribute to the exacerbated inflammatory reactions observed in ALS mice.}, }
@article {pmid31171724, year = {2019}, author = {Ryan, JJ and Sprunger, ML and Holthaus, K and Shorter, J and Jackrel, ME}, title = {Engineered protein disaggregases mitigate toxicity of aberrant prion-like fusion proteins underlying sarcoma.}, journal = {The Journal of biological chemistry}, volume = {294}, number = {29}, pages = {11286-11296}, pmid = {31171724}, issn = {1083-351X}, support = {R01 GM099836/GM/NIGMS NIH HHS/United States ; R35 GM128772/GM/NIGMS NIH HHS/United States ; S10 OD021629/OD/NIH HHS/United States ; }, mesh = {Cell Nucleus/metabolism ; Heat-Shock Proteins/metabolism ; Humans ; Oncogene Proteins, Fusion/genetics/metabolism ; Prion Proteins/*metabolism ; *Protein Engineering ; Proto-Oncogene Protein c-fli-1/genetics ; RNA-Binding Protein EWS/genetics ; RNA-Binding Protein FUS/genetics ; Sarcoma/*metabolism ; Soft Tissue Neoplasms/*metabolism ; Transcription Factor CHOP/metabolism ; }, abstract = {FUS and EWSR1 are RNA-binding proteins with prion-like domains (PrLDs) that aggregate in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The FUS and EWSR1 genes are also prone to chromosomal translocation events, which result in aberrant fusions between portions of the PrLDs of FUS and EWSR1 and the transcription factors CHOP and FLI. The resulting fusion proteins, FUS-CHOP and EWS-FLI, drive aberrant transcriptional programs that underpin liposarcoma and Ewing's sarcoma, respectively. The translocated PrLDs alter the expression profiles of these proteins and promote their phase separation and aggregation. Here, we report the development of yeast models of FUS-CHOP and EWS-FLI toxicity and aggregation. These models recapitulated several salient features of sarcoma patient cells harboring the FUS-CHOP and EWS-FLI translocations. To reverse FUS and EWSR1 aggregation, we have explored Hsp104, a hexameric AAA+ protein disaggregase from yeast. Previously, we engineered potentiated Hsp104 variants to suppress the proteotoxicity, aggregation, and mislocalization of FUS and other proteins that aggregate in ALS/FTD and Parkinson's disease. Potentiated Hsp104 variants that robustly suppressed FUS toxicity and aggregation also suppressed the toxicity and aggregation of FUS-CHOP and EWS-FLI. We suggest that these new yeast models are powerful platforms for screening for modulators of FUS-CHOP and EWS-FLI phase separation. Moreover, Hsp104 variants might be employed to combat the toxicity and phase separation of aberrant fusion proteins involved in sarcoma.}, }
@article {pmid31193360, year = {2019}, author = {Subaraja, M and Vanisree, AJ}, title = {Counter effects of Asiaticosids-D through putative neurotransmission on rotenone induced cerebral ganglionic injury in Lumbricus terrestris.}, journal = {IBRO reports}, volume = {6}, number = {}, pages = {160-175}, pmid = {31193360}, issn = {2451-8301}, abstract = {Asiaticoside-D (AD) was shown to efficacy of ganglionic degenerated Lumbricus terrestris as a pioneering observation in our earlier research. Though, extract molecular mechanisms of AD for degenerative diseases (DDs) remains largely unknown. We investigated the neuroprotective effects of AD against ROT in cerebral ganglions (CGs) of degenerative L. terrestris. Worms were exposed to 0.4 ppm ROT for 7 days were subjected to co- treatment with 15 ppm of AD. After, CGs was removed. The levels oxidant, non-antioxidant, antioxidant status, ganglioside, ceramide and ceramide glycanase (CGase) were estimated. The m-RNA levels of dopamine transporter (DAT), octopamine transporter (OAT), innexins-9 (inx-9), ionotropic glutamate receptor 3 (iGlu3), heat shock proteins (hsp70), XPRLamide neuropeptide precursor, tyramine beta-hydroxylase (tbh-1) and β- adrenergic receptor kinase-2 (β-ARK2-3) by semi-qRT- PCR. The expression pattern of tyramine beta hydroxylase (TBH), glutamate receptor (iGluR), serotonin transporter (SERT), dopamine transporters (DAT), nerve growth factors (NGF), cytochrome C oxidase (COC), NADH dehydogenase subunit-1 (ND-1), neurotrophin receptor p75 (p75NTR), neuronal nitric oxiside synthase (nNOs) interleukin 1- beta (IL1-β) and tumor necrosis factor alpha (TNF-α) by western blotting. Glutaminergic, serotogenic and dopaminergic toxicity variations were also performed. The levels of oxidant, non-antioxidant, antioxidant status, lipids, proteins and m-RNAs were significantly altered (p < 0.001) on ROT-induced (group II) and their levels were significantly changes (p < 0.05) by ROT+AD in CGs. The sensitive study plan concluded the neuroprotective effects of AD against ROT induced degeneration in worms and suggest that the AD deserves future studies for its use as an effective alternative medicine that could minimize the morbidity of ganglionic degenerative diseases patients.}, }
@article {pmid31382568, year = {2019}, author = {Apolloni, S and Caputi, F and Pignataro, A and Amadio, S and Fabbrizio, P and Ammassari-Teule, M and Volonté, C}, title = {Histamine Is an Inducer of the Heat Shock Response in SOD1-G93A Models of ALS.}, journal = {International journal of molecular sciences}, volume = {20}, number = {15}, pages = {}, pmid = {31382568}, issn = {1422-0067}, support = {Flagship Project NanoMAX (B81J13000310005)//Ministero dell'Istruzione, dell'Universit&#xe0; e della Ricerca/ ; }, mesh = {Amyotrophic Lateral Sclerosis ; Animals ; Astrocytes/drug effects/pathology ; Cell Death/drug effects ; Dendritic Spines/drug effects/genetics ; Disease Models, Animal ; Endoplasmic Reticulum Chaperone BiP ; Heat-Shock Response/*drug effects/genetics ; Histamine/*pharmacology ; Humans ; Mice ; Microglia/metabolism/pathology ; Motor Neurons/*drug effects/pathology ; Mutation ; Neuroglia/drug effects/pathology ; Spinal Cord/drug effects/pathology ; Superoxide Dismutase-1/*genetics ; }, abstract = {(1) Background: Amyotrophic lateral sclerosis (ALS) is a multifactorial non-cell autonomous disease where activation of microglia and astrocytes largely contributes to motor neurons death. Heat shock proteins have been demonstrated to promote neuronal survival and exert a strong anti-inflammatory action in glia. Having previously shown that the pharmacological increase of the histamine content in the central nervous system (CNS) of SOD1-G93A mice decreases neuroinflammation, reduces motor neuron death, and increases mice life span, here we examined whether this effect could be mediated by an enhancement of the heat shock response. (2) Methods: Heat shock protein expression was analyzed in vitro and in vivo. Histamine was provided to primary microglia and NSC-34 motor neurons expressing the SOD1-G93A mutation. The brain permeable histamine precursor histidine was chronically administered to symptomatic SOD1-G93A mice. Spine density was measured by Golgi-staining in motor cortex of histidine-treated SOD1-G93A mice. (3) Results: We demonstrate that histamine activates the heat shock response in cultured SOD1-G93A microglia and motor neurons. In SOD1-G93A mice, histidine augments the protein content of GRP78 and Hsp70 in spinal cord and cortex, where the treatment also rescues type I motor neuron dendritic spine loss. (4) Conclusion: Besides the established histaminergic neuroprotective and anti-inflammatory effects, the induction of the heat shock response in the SOD1-G93A model by histamine confirms the importance of this pathway in the search for successful therapeutic solutions to treat ALS.}, }
@article {pmid31427919, year = {2019}, author = {Cristofani, R and Rusmini, P and Galbiati, M and Cicardi, ME and Ferrari, V and Tedesco, B and Casarotto, E and Chierichetti, M and Messi, E and Piccolella, M and Carra, S and Crippa, V and Poletti, A}, title = {The Regulation of the Small Heat Shock Protein B8 in Misfolding Protein Diseases Causing Motoneuronal and Muscle Cell Death.}, journal = {Frontiers in neuroscience}, volume = {13}, number = {}, pages = {796}, pmid = {31427919}, issn = {1662-4548}, abstract = {Misfolding protein diseases are a wide class of disorders in which the aberrantly folded protein aggregates accumulate in affected cells. In the brain and in the skeletal muscle, misfolded protein accumulation induces a variety of cell dysfunctions that frequently lead to cell death. In motoneuron diseases (MNDs), misfolded proteins accumulate primarily in motoneurons, glial cells and/or skeletal muscle cells, altering motor function. The deleterious effects of misfolded proteins can be counteracted by the activity of the protein quality control (PQC) system, composed of chaperone proteins and degradative systems. Here, we focus on a PQC system component: heat shock protein family B (small) member 8 (HSPB8), a chaperone induced by harmful stressful events, including proteotoxicity. In motoneuron and muscle cells, misfolded proteins activate HSPB8 transcription and enhance HSPB8 levels, which contributes to prevent aggregate formation and their harmful effects. HSPB8 acts not only as a chaperone, but also facilitates the autophagy process, to enable the efficient clearance of the misfolded proteins. HSPB8 acts as a dimer bound to the HSP70 co-chaperone BAG3, a scaffold protein that is also capable of binding to HSP70 (associated with the E3-ligase CHIP) and dynein. When this complex is formed, it is transported by dynein to the microtubule organization center (MTOC), where aggresomes are formed. Here, misfolded proteins are engulfed into nascent autophagosomes to be degraded via the chaperone-assisted selective autophagy (CASA). When CASA is insufficient or impaired, HSP70 and CHIP associate with an alternative co-chaperone, BAG1, which routes misfolded proteins to the proteasome for degradation. The finely tuned equilibrium between proteasome and CASA activity is thought to be crucial for maintaining the functional cell homeostasis during proteotoxic stresses, which in turn is essential for cell survival. This fine equilibrium seems to be altered in MNDs, like Amyotrophic lateral sclerosis (ALS) and spinal and bulbar muscular atrophy (SBMA), contributing to the onset and the progression of disease. Here, we will review how misfolded proteins may affect the PQC system and how the proper activity of this system can be restored by boosting or regulating HSPB8 activity, with the aim to ameliorate disease progression in these two fatal MNDs.}, }
@article {pmid31476356, year = {2019}, author = {Lyon, MS and Milligan, C}, title = {Extracellular heat shock proteins in neurodegenerative diseases: New perspectives.}, journal = {Neuroscience letters}, volume = {711}, number = {}, pages = {134462}, doi = {10.1016/j.neulet.2019.134462}, pmid = {31476356}, issn = {1872-7972}, mesh = {Animals ; Heat-Shock Proteins/*metabolism ; Humans ; Neurodegenerative Diseases/*metabolism ; }, abstract = {One pathological hallmark of neurodegenerative diseases and CNS trauma is accumulation of insoluble, hydrophobic molecules and protein aggregations found both within and outside cells. These may be the consequences of an inadequate or overburdened cellular response to stresses resulting from potentially toxic changes in extra- and intracellular environments. The upregulated expression of heat shock proteins (HSPs) is one example of a highly conserved cellular response to both internal and external stress. Intracellularly these proteins act as chaperones, playing vital roles in the folding of nascent polypeptides, the translocation of proteins between subcellular locations, and the disaggregation of misfolded or aggregated proteins in an attempt to maintain cellular proteostasis during both homeostatic and stressful conditions. While the predominant study of the HSPs has focused on their intracellular chaperone functions, it remains unclear if all neuronal populations can mount a complete stress response. Alternately, it is now well established that some members of this family of proteins can be secreted by nearby, non-neuronal cells to act in the extracellular environment. This review addresses the current literature detailing the use of exogenous and extracellular HSPs in the treatment of cellular and animal models of neurodegenerative disease. These findings offer a new measure of therapeutic potential to the HSPs, but obstacles must be overcome before they can be efficiently used in a clinical setting.}, }
@article {pmid31521619, year = {2019}, author = {Zhang, C and Liang, W and Wang, H and Yang, Y and Wang, T and Wang, S and Wang, X and Wang, Y and Feng, H}, title = {γ-Oryzanol mitigates oxidative stress and prevents mutant SOD1-Related neurotoxicity in Drosophila and cell models of amyotrophic lateral sclerosis.}, journal = {Neuropharmacology}, volume = {160}, number = {}, pages = {107777}, doi = {10.1016/j.neuropharm.2019.107777}, pmid = {31521619}, issn = {1873-7064}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/metabolism ; Animals ; Antioxidants/pharmacology ; Cell Line ; Disease Models, Animal ; Drosophila ; Drosophila Proteins/genetics/metabolism ; Humans ; Male ; Mice ; Motor Neurons/drug effects ; Mutation ; Neuroprotective Agents/administration &amp; dosage/*pharmacology ; Oxidative Stress/*drug effects ; Phenylpropionates/administration &amp; dosage/*pharmacology ; Superoxide Dismutase/genetics/metabolism ; Superoxide Dismutase-1/*genetics/metabolism ; }, abstract = {Oxidative stress plays a critical role in mutant copper/zinc superoxide dismutase 1 (SOD1)-linked amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease characterized by selective loss of motor neurons. Thus, an anti-oxidative stress remedy might be a promising means for the treatment of ALS. The aim of the present study is to investigate the neuroprotective effects of γ-oryzanol (Orz) and elucidate its relevant molecular mechanisms in mutant hSOD1-linked Drosophila and cell models of ALS. Orz treatment provided neuroprotection in flies with expression of hSOD1-G85R in motor neurons, as demonstrated by the prolonged survival, improvement of motor deficits, reduced oxidative damage and regulated redox homeostasis when compared with those in controls. Moreover, Orz significantly decreased neuronal apoptosis and upregulated the nuclear factor erythroid 2-related factor 2 (Nrf2)/glutamate-cysteine ligase catalytic subunit (GCLC) antioxidant pathway via activating Akt in hSOD1-G93A-expressing NSC-34 cells. In addition, our results showed that both in vivo and in vitro, Akt served as an upstream regulator of signal transducers and activators of transcription (Stat) 3 stimulated by Orz, which further increased the level of another anti-oxidative stress factor heat-shock protein 70 (HSP70). Altogether, these findings provide evidence that Orz has potential neuroprotective effects that may be beneficial in the treatment of ALS disease with SOD1 mutations.}, }
@article {pmid31738976, year = {2020}, author = {Forcella, M and Lau, P and Oldani, M and Melchioretto, P and Bogni, A and Gribaldo, L and Fusi, P and Urani, C}, title = {Neuronal specific and non-specific responses to cadmium possibly involved in neurodegeneration: A toxicogenomics study in a human neuronal cell model.}, journal = {Neurotoxicology}, volume = {76}, number = {}, pages = {162-173}, doi = {10.1016/j.neuro.2019.11.002}, pmid = {31738976}, issn = {1872-9711}, mesh = {Cadmium/*toxicity ; Cell Line, Tumor ; Gene Expression/*drug effects ; Humans ; Metallothionein/metabolism ; Neurons/*drug effects/*metabolism ; Signal Transduction/drug effects ; Toxicogenetics ; }, abstract = {Epidemiological data have linked cadmium exposure to neurotoxicity and to neurodegenerative diseases (e.g., Alzheimer's and Parkinson's disease), and to increased risk of developing ALS. Even though the brain is not a primary target organ, this metal can bypass the blood brain barrier, thus exerting its toxic effects. The coordination chemistry of cadmium is of strong biological relevance, as it resembles to zinc(II) and calcium(II), two ions crucial for neuronal signaling. A toxicogenomics approach applied to a neuronal human model (SH-SY5Y cells) exposed to cadmium (10 and 20 μM) allowed the identification of early deregulated genes and altered processes, and the discrimination between neuronal-specific and unspecific responses as possible triggers of neurodegeneration. Cadmium confirmed its recognized carcinogenicity even on neuronal cells by activating the p53 signaling pathway and genes involved in tumor initiation and cancer cell proliferation, and by down-regulating genes coding for tumor suppressors and for DNA repair enzymes. Two cadmium-induced stress responses were observed: the activation of different members of the heat shock family, as a mechanism to restore protein folding in response to proteotoxicity, and the activation of metallothioneins (MTs), involved in zinc and copper homeostasis, protection against metal toxicity and oxidative damage. Perturbed function of essential metals is suggested by the mineral absorption pathway, with MTs, HMOX1, ZnT-1, and Ferritin genes highly up-regulated. Cadmium interferes also with Ca[2+] regulation as S100A2 is one of the top up-regulated genes, coding for a highly specialized family of regulatory Ca[2+]-binding proteins. Other neuronal-related functions altered in SH-SY5Y cells by cadmium are microtubules dynamics, microtubules motor-based proteins and neuroprotection by down-regulation of NEK3, KIF15, and GREM2 genes, respectively.}, }
@article {pmid31768050, year = {2019}, author = {Farhan, SMK and Howrigan, DP and Abbott, LE and Klim, JR and Topp, SD and Byrnes, AE and Churchhouse, C and Phatnani, H and Smith, BN and Rampersaud, E and Wu, G and Wuu, J and Shatunov, A and Iacoangeli, A and Al Khleifat, A and Mordes, DA and Ghosh, S and ,  and ,  and ,  and ,  and Eggan, K and Rademakers, R and McCauley, JL and Schüle, R and Züchner, S and Benatar, M and Taylor, JP and Nalls, M and Gotkine, M and Shaw, PJ and Morrison, KE and Al-Chalabi, A and Traynor, B and Shaw, CE and Goldstein, DB and Harms, MB and Daly, MJ and Neale, BM}, title = {Exome sequencing in amyotrophic lateral sclerosis implicates a novel gene, DNAJC7, encoding a heat-shock protein.}, journal = {Nature neuroscience}, volume = {22}, number = {12}, pages = {1966-1974}, pmid = {31768050}, issn = {1546-1726}, support = {U2C TR002818/TR/NCATS NIH HHS/United States ; MR/L501529/1/MRC_/Medical Research Council/United Kingdom ; U54 NS092091/NS/NINDS NIH HHS/United States ; G0600974/MRC_/Medical Research Council/United Kingdom ; G0500289/MRC_/Medical Research Council/United Kingdom ; SMITH/APR16/847-791/MNDA_/Motor Neurone Disease Association/United Kingdom ; G0900688/MRC_/Medical Research Council/United Kingdom ; MC_PC_17115/MRC_/Medical Research Council/United Kingdom ; U01 MH115727/MH/NIMH NIH HHS/United States ; G0900635/MRC_/Medical Research Council/United Kingdom ; ALCHALABI-DOBSON/APR14/829-791/MNDA_/Motor Neurone Disease Association/United Kingdom ; MR/L021803/1/MRC_/Medical Research Council/United Kingdom ; G1100695/MRC_/Medical Research Council/United Kingdom ; MR/R024804/1/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; MRF-060-0003-RG-SMITH/MRF_/MRF_/United Kingdom ; SHAW/NOV14/985-797/MNDA_/Motor Neurone Disease Association/United Kingdom ; }, mesh = {Amyotrophic Lateral Sclerosis/*genetics ; Case-Control Studies ; Exome/*genetics ; Female ; Genetic Predisposition to Disease/*genetics ; Genetic Variation/genetics ; Heat-Shock Proteins/*genetics ; Humans ; Male ; Molecular Chaperones/*genetics ; }, abstract = {To discover novel genes underlying amyotrophic lateral sclerosis (ALS), we aggregated exomes from 3,864 cases and 7,839 ancestry-matched controls. We observed a significant excess of rare protein-truncating variants among ALS cases, and these variants were concentrated in constrained genes. Through gene level analyses, we replicated known ALS genes including SOD1, NEK1 and FUS. We also observed multiple distinct protein-truncating variants in a highly constrained gene, DNAJC7. The signal in DNAJC7 exceeded genome-wide significance, and immunoblotting assays showed depletion of DNAJC7 protein in fibroblasts in a patient with ALS carrying the p.Arg156Ter variant. DNAJC7 encodes a member of the heat-shock protein family, HSP40, which, along with HSP70 proteins, facilitates protein homeostasis, including folding of newly synthesized polypeptides and clearance of degraded proteins. When these processes are not regulated, misfolding and accumulation of aberrant proteins can occur and lead to protein aggregation, which is a pathological hallmark of neurodegeneration. Our results highlight DNAJC7 as a novel gene for ALS.}, }
@article {pmid31852729, year = {2020}, author = {Clark, EM and Nonarath, HJT and Bostrom, JR and Link, BA}, title = {Establishment and validation of an endoplasmic reticulum stress reporter to monitor zebrafish ATF6 activity in development and disease.}, journal = {Disease models &amp; mechanisms}, volume = {13}, number = {1}, pages = {}, pmid = {31852729}, issn = {1754-8411}, support = {R01 EY029267/EY/NEI NIH HHS/United States ; T32 EY014537/EY/NEI NIH HHS/United States ; TL1 TR001437/TR/NCATS NIH HHS/United States ; }, mesh = {Activating Transcription Factor 6/genetics/*physiology ; Amyotrophic Lateral Sclerosis/genetics ; Animals ; Disease Models, Animal ; *Endoplasmic Reticulum Stress ; Neurodegenerative Diseases/genetics ; Signal Transduction/physiology ; Transgenes ; Unfolded Protein Response ; Zebrafish/*embryology ; }, abstract = {Induction of endoplasmic reticulum (ER) stress is associated with diverse developmental and degenerative diseases. Modified ER homeostasis causes activation of conserved stress pathways at the ER called the unfolded protein response (UPR). ATF6 is a transcription factor activated during ER stress as part of a coordinated UPR. ATF6 resides at the ER and, upon activation, is transported to the Golgi apparatus, where it is cleaved by proteases to create an amino-terminal cytoplasmic fragment (ATF6f). ATF6f translocates to the nucleus to activate transcriptional targets. Here, we describe the establishment and validation of zebrafish reporter lines for ATF6 activity. These transgenic lines are based on a defined and multimerized ATF6 consensus site, which drives either eGFP or destabilized eGFP, enabling dynamic study of ATF6 activity during development and disease. The results show that the reporter is specific for the ATF6 pathway, active during development and induced in disease models known to engage UPR. Specifically, during development, ATF6 activity is highest in the lens, skeletal muscle, fins and gills. The reporter is also activated by common chemical inducers of ER stress, including tunicamycin, thapsigargin and brefeldin A, as well as by heat shock. In models for amyotrophic lateral sclerosis and cone dystrophy, ATF6 reporter expression is induced in spinal cord interneurons or photoreceptors, respectively, suggesting a role for ATF6 response in multiple neurodegenerative diseases. Collectively our results show that these ATF6 reporters can be used to monitor ATF6 activity changes throughout development and in zebrafish models of disease.This article has an associated First Person interview with the first author of the paper.}, }
@article {pmid31857710, year = {2020}, author = {Farhan, SMK and Howrigan, DP and Abbott, LE and Klim, JR and Topp, SD and Byrnes, AE and Churchhouse, C and Phatnani, H and Smith, BN and Rampersaud, E and Wu, G and Wuu, J and Shatunov, A and Iacoangeli, A and Khleifat, AA and Mordes, DA and Ghosh, S and ,  and ,  and ,  and ,  and Eggan, K and Rademakers, R and McCauley, JL and Schüle, R and Züchner, S and Benatar, M and Taylor, JP and Nalls, M and Gotkine, M and Shaw, PJ and Morrison, KE and Al-Chalabi, A and Traynor, B and Shaw, CE and Goldstein, DB and Harms, MB and Daly, MJ and Neale, BM}, title = {Publisher Correction: Exome sequencing in amyotrophic lateral sclerosis implicates a novel gene, DNAJC7, encoding a heat-shock protein.}, journal = {Nature neuroscience}, volume = {23}, number = {2}, pages = {295}, doi = {10.1038/s41593-019-0570-5}, pmid = {31857710}, issn = {1546-1726}, support = {MR/L501529/1/MRC_/Medical Research Council/United Kingdom ; U2C TR002818/TR/NCATS NIH HHS/United States ; }, abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.}, }
@article {pmid31900865, year = {2020}, author = {Kuta, R and Larochelle, N and Fernandez, M and Pal, A and Minotti, S and Tibshirani, M and St Louis, K and Gentil, BJ and Nalbantoglu, JN and Hermann, A and Durham, HD}, title = {Depending on the stress, histone deacetylase inhibitors act as heat shock protein co-inducers in motor neurons and potentiate arimoclomol, exerting neuroprotection through multiple mechanisms in ALS models.}, journal = {Cell stress &amp; chaperones}, volume = {25}, number = {1}, pages = {173-191}, pmid = {31900865}, issn = {1466-1268}, mesh = {Amyotrophic Lateral Sclerosis/*drug therapy/genetics ; Animals ; Cells, Cultured ; HSP70 Heat-Shock Proteins/metabolism ; Heat-Shock Proteins/*drug effects/metabolism ; Heat-Shock Response/drug effects ; Histone Deacetylase Inhibitors/pharmacology ; Hydroxylamines/*pharmacology ; Mice ; Motor Neurons/*drug effects/metabolism ; Spinal Cord/*drug effects/metabolism ; Transcriptional Activation/drug effects ; Up-Regulation/drug effects ; }, abstract = {Upregulation of heat shock proteins (HSPs) is an approach to treatment of neurodegenerative disorders with impaired proteostasis. Many neurons, including motor neurons affected in amyotrophic lateral sclerosis (ALS), are relatively resistant to stress-induced upregulation of HSPs. This study demonstrated that histone deacetylase (HDAC) inhibitors enable the heat shock response in cultured spinal motor neurons, in a stress-dependent manner, and can improve the efficacy of HSP-inducing drugs in murine spinal cord cultures subjected to thermal or proteotoxic stress. The effect of particular HDAC inhibitors differed with the stress paradigm. The HDAC6 (class IIb) inhibitor, tubastatin A, acted as a co-inducer of Hsp70 (HSPA1A) expression with heat shock, but not with proteotoxic stress induced by expression of mutant SOD1 linked to familial ALS. Certain HDAC class I inhibitors (the pan inhibitor, SAHA, or the HDAC1/3 inhibitor, RGFP109) were HSP co-inducers comparable to the hydroxyamine arimoclomol in response to proteotoxic stress, but not thermal stress. Regardless, stress-induced Hsp70 expression could be enhanced by combining an HDAC inhibitor with either arimoclomol or with an HSP90 inhibitor that constitutively induced HSPs. HDAC inhibition failed to induce Hsp70 in motor neurons expressing ALS-linked mutant FUS, in which the heat shock response was suppressed; yet SAHA, RGFP109, and arimoclomol did reduce loss of nuclear FUS, a disease hallmark, and HDAC inhibition rescued the DNA repair response in iPSC-derived motor neurons carrying the FUS[P525L]mutation, pointing to multiple mechanisms of neuroprotection by both HDAC inhibiting drugs and arimoclomol.}, }
@article {pmid32084385, year = {2020}, author = {Abo-Rady, M and Kalmbach, N and Pal, A and Schludi, C and Janosch, A and Richter, T and Freitag, P and Bickle, M and Kahlert, AK and Petri, S and Stefanov, S and Glass, H and Staege, S and Just, W and Bhatnagar, R and Edbauer, D and Hermann, A and Wegner, F and Sterneckert, JL}, title = {Knocking out C9ORF72 Exacerbates Axonal Trafficking Defects Associated with Hexanucleotide Repeat Expansion and Reduces Levels of Heat Shock Proteins.}, journal = {Stem cell reports}, volume = {14}, number = {3}, pages = {390-405}, pmid = {32084385}, issn = {2213-6711}, mesh = {Amyotrophic Lateral Sclerosis/genetics/pathology ; Apoptosis/drug effects ; Axons/drug effects/*metabolism ; Benzhydryl Compounds/pharmacology ; C9orf72 Protein/*genetics/metabolism ; Cell Differentiation/drug effects ; Cytoplasmic Granules/drug effects/metabolism ; DNA Repeat Expansion/*genetics ; Gain of Function Mutation/genetics ; *Gene Knockout Techniques ; HSP40 Heat-Shock Proteins/*metabolism ; HSP70 Heat-Shock Proteins/*metabolism ; Humans ; Induced Pluripotent Stem Cells/drug effects/metabolism ; Models, Biological ; Motor Neurons/drug effects/metabolism/pathology ; Nerve Degeneration/pathology ; Pyrrolidinones/pharmacology ; Transcriptome/genetics ; }, abstract = {In amyotrophic lateral sclerosis (ALS) motor neurons (MNs) undergo dying-back, where the distal axon degenerates before the soma. The hexanucleotide repeat expansion (HRE) in C9ORF72 is the most common genetic cause of ALS, but the mechanism of pathogenesis is largely unknown with both gain- and loss-of-function mechanisms being proposed. To better understand C9ORF72-ALS pathogenesis, we generated isogenic induced pluripotent stem cells. MNs with HRE in C9ORF72 showed decreased axonal trafficking compared with gene corrected MNs. However, knocking out C9ORF72 did not recapitulate these changes in MNs from healthy controls, suggesting a gain-of-function mechanism. In contrast, knocking out C9ORF72 in MNs with HRE exacerbated axonal trafficking defects and increased apoptosis as well as decreased levels of HSP70 and HSP40, and inhibition of HSPs exacerbated ALS phenotypes in MNs with HRE. Therefore, we propose that the HRE in C9ORF72 induces ALS pathogenesis via a combination of gain- and loss-of-function mechanisms.}, }
@article {pmid32087285, year = {2020}, author = {Luotti, S and Pasetto, L and Porcu, L and Torri, V and Elezgarai, SR and Pantalone, S and Filareti, M and Corbo, M and Lunetta, C and Mora, G and Bonetto, V}, title = {Diagnostic and prognostic values of PBMC proteins in amyotrophic lateral sclerosis.}, journal = {Neurobiology of disease}, volume = {139}, number = {}, pages = {104815}, doi = {10.1016/j.nbd.2020.104815}, pmid = {32087285}, issn = {1095-953X}, mesh = {Adult ; Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/*diagnosis/metabolism ; Biomarkers/metabolism ; Case-Control Studies ; DNA-Binding Proteins/metabolism ; Female ; Humans ; Leukocytes, Mononuclear/*metabolism ; Male ; Middle Aged ; Motor Neurons/metabolism ; Peptidylprolyl Isomerase/metabolism ; Prognosis ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease for which there are no validated biomarkers. Previous exploratory studies have identified a panel of candidate protein biomarkers in peripheral blood mononuclear cells (PBMCs) that include peptidyl-prolyl cis-trans isomerase A (PPIA), heat shock cognate protein 71 kDa (HSC70), heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2B1) and TDP-43. It has also been found that PPIA plays a key role in the assembly and dynamics of ribonucleoprotein (RNP) complexes and interacts with TDP-43. Its absence accelerates disease progression in a SOD1 mouse model of ALS, and low levels of PPIA in PBMCs are associated with early-onset ALS. However, the diagnostic and prognostic values of PPIA and the other candidate protein biomarkers have not been established. We analyzed the PBMC proteins in a well-characterized cohort of ALS patients (n=93), healthy individuals (n=104) and disease controls (n=111). We used a highly controlled sample processing procedure that implies two-step differential detergent fractionation. We found that the levels of the selected PBMC proteins in the soluble and insoluble fraction, combined, have a high discriminatory power for distinguishing ALS from controls, with PPIA, hnRNPA2B1 and TDP-43 being the proteins most closely associated with ALS. We also found a shift toward increased protein partitioning in the insoluble fraction in ALS and this correlated with a worse disease phenotype. In particular, low PPIA soluble levels were associated with six months earlier death. In conclusion, PPIA is a disease modifier with prognostic potential. PBMC proteins indicative of alterations in protein and RNA homeostasis are promising biomarkers of ALS, for diagnosis, prognosis and patient stratification.}, }
@article {pmid32284329, year = {2020}, author = {Serlidaki, D and van Waarde, MAWH and Rohland, L and Wentink, AS and Dekker, SL and Kamphuis, MJ and Boertien, JM and Brunsting, JF and Nillegoda, NB and Bukau, B and Mayer, MP and Kampinga, HH and Bergink, S}, title = {Functional diversity between HSP70 paralogs caused by variable interactions with specific co-chaperones.}, journal = {The Journal of biological chemistry}, volume = {295}, number = {21}, pages = {7301-7316}, pmid = {32284329}, issn = {1083-351X}, mesh = {Amino Acid Substitution ; Cell Line, Tumor ; HEK293 Cells ; HSP110 Heat-Shock Proteins/*chemistry/genetics ; HSP70 Heat-Shock Proteins/*chemistry/genetics ; Homeodomain Proteins/*chemistry/genetics ; Humans ; Mutation, Missense ; *Protein Aggregation, Pathological ; Superoxide Dismutase-1/*chemistry/genetics ; Tumor Suppressor Proteins/*chemistry/genetics ; }, abstract = {Heat shock protein 70 (HSP70) chaperones play a central role in protein quality control and are crucial for many cellular processes, including protein folding, degradation, and disaggregation. Human HSP70s compose a family of 13 members that carry out their functions with the aid of even larger families of co-chaperones. A delicate interplay between HSP70s and co-chaperone recruitment is thought to determine substrate fate, yet it has been generally assumed that all Hsp70 paralogs have similar activities and are largely functionally redundant. However, here we found that when expressed in human cells, two highly homologous HSP70s, HSPA1A and HSPA1L, have opposing effects on cellular handling of various substrates. For example, HSPA1A reduced aggregation of the amyotrophic lateral sclerosis-associated protein variant superoxide dismutase 1 (SOD1)-A4V, whereas HSPA1L enhanced its aggregation. Intriguingly, variations in the substrate-binding domain of these HSP70s did not play a role in this difference. Instead, we observed that substrate fate is determined by differential interactions of the HSP70s with co-chaperones. Whereas most co-chaperones bound equally well to these two HSP70s, Hsp70/Hsp90-organizing protein (HOP) preferentially bound to HSPA1L, and the Hsp110 nucleotide-exchange factor HSPH2 preferred HSPA1A. The role of HSPH2 was especially crucial for the HSPA1A-mediated reduction in SOD1-A4V aggregation. These findings reveal a remarkable functional diversity at the level of the cellular HSP70s and indicate that this diversity is defined by their affinities for specific co-chaperones such as HSPH2.}, }
@article {pmid32289328, year = {2020}, author = {Upadhya, R and Zingg, W and Shetty, S and Shetty, AK}, title = {Astrocyte-derived extracellular vesicles: Neuroreparative properties and role in the pathogenesis of neurodegenerative disorders.}, journal = {Journal of controlled release : official journal of the Controlled Release Society}, volume = {323}, number = {}, pages = {225-239}, pmid = {32289328}, issn = {1873-4995}, support = {R01 NS106907/NS/NINDS NIH HHS/United States ; }, mesh = {*Alzheimer Disease ; Astrocytes ; *Extracellular Vesicles ; Humans ; *Neurodegenerative Diseases ; Vascular Endothelial Growth Factor A ; }, abstract = {Extracellular vesicles (EVs) released by neural cells play an essential role in brain homeostasis and the crosstalk between neural cells and the periphery. EVs are diverse, nano-sized vesicles, which transport proteins, nucleic acids, and lipids between cells over short and long expanses and hence are proficient for modulating the target cells. EVs released from neural cells are implicated in synaptic plasticity, neuron-glia interface, neuroprotection, neuroregeneration, and the dissemination of neuropathological molecules. This review confers the various properties of EVs secreted by astrocytes and their potential role in health and disease with a focus on evolving concepts. Naïve astrocytes shed EVs containing a host of neuroprotective compounds, which include fibroblast growth factor-2, vascular endothelial growth factor, and apolipoprotein-D. Stimulated astrocytes secrete EVs with neuroprotective molecules including heat shock proteins, synapsin 1, unique microRNAs, and glutamate transporters. Well-characterized astrocyte-derived EVs (ADEVs) generated in specific culture conditions and ADEVs that are engineered to carry the desired miRNAs or proteins are likely useful for treating brain injury and neurogenerative diseases. On the other hand, in conditions such as Alzheimer's disease (AD), stroke, Parkinson's disease, Amyotrophic lateral sclerosis (ALS), and other neuroinflammatory conditions, EVs released by activated astrocytes appear to mediate or exacerbate the pathological processes. The examples include ADEVs spreading the dysregulated complement system in AD, mediating motoneuron toxicity in ALS, and stimulating peripheral leukocyte migration into the brain in inflammatory conditions. Strategies restraining the release of EVs by activated astrocytes or modulating the composition of ADEVs are likely beneficial for treating neurodegenerative diseases. Also, periodic analyses of ADEVs in the blood is useful for detecting astrocyte-specific biomarkers in different neurological conditions and for monitoring disease progression and remission with distinct therapeutic approaches.}, }
@article {pmid32297211, year = {2020}, author = {Sinnige, T and Yu, A and Morimoto, RI}, title = {Challenging Proteostasis: Role of the Chaperone Network to Control Aggregation-Prone Proteins in Human Disease.}, journal = {Advances in experimental medicine and biology}, volume = {1243}, number = {}, pages = {53-68}, pmid = {32297211}, issn = {0065-2598}, support = {P01 AG054407/AG/NIA NIH HHS/United States ; R37 AG026647/AG/NIA NIH HHS/United States ; R56 AG059579/AG/NIA NIH HHS/United States ; RF1 AG057296/AG/NIA NIH HHS/United States ; }, mesh = {Animals ; Humans ; Molecular Chaperones/*metabolism ; Neurodegenerative Diseases/*metabolism/*pathology/prevention &amp; control ; Protein Aggregation, Pathological/*prevention &amp; control ; *Proteostasis ; }, abstract = {Protein homeostasis (Proteostasis) is essential for correct and efficient protein function within the living cell. Among the critical components of the Proteostasis Network (PN) are molecular chaperones that serve widely in protein biogenesis under physiological conditions, and prevent protein misfolding and aggregation enhanced by conditions of cellular stress. For Alzheimer's, Parkinson's, Huntington's diseases and ALS, multiple classes of molecular chaperones interact with the highly aggregation-prone proteins amyloid-β, tau, α-synuclein, huntingtin and SOD1 to influence the course of proteotoxicity associated with these neurodegenerative diseases. Accordingly, overexpression of molecular chaperones and induction of the heat shock response have been shown to be protective in a wide range of animal models of these diseases. In contrast, for cancer cells the upregulation of chaperones has the undesirable effect of promoting cellular survival and tumor growth by stabilizing mutant oncoproteins. In both situations, physiological levels of molecular chaperones eventually become functionally compromised by the persistence of misfolded substrates, leading to a decline in global protein homeostasis and the dysregulation of diverse cellular pathways. The phenomenon of chaperone competition may underlie the broad pathology observed in aging and neurodegenerative diseases, and restoration of physiological protein homeostasis may be a suitable therapeutic avenue for neurodegeneration as well as for cancer.}, }
@article {pmid32324341, year = {2020}, author = {Montibeller, L and Tan, LY and Kim, JK and Paul, P and de Belleroche, J}, title = {Tissue-selective regulation of protein homeostasis and unfolded protein response signalling in sporadic ALS.}, journal = {Journal of cellular and molecular medicine}, volume = {24}, number = {11}, pages = {6055-6069}, pmid = {32324341}, issn = {1582-4934}, mesh = {Adult ; Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/genetics/*metabolism/pathology ; Case-Control Studies ; Endoplasmic Reticulum Stress/genetics ; Frontotemporal Dementia/genetics/pathology ; Gene Expression Regulation ; HSP40 Heat-Shock Proteins/genetics/metabolism ; Heat Shock Transcription Factors/metabolism ; Heat-Shock Response/genetics ; Humans ; Middle Aged ; Models, Biological ; Molecular Chaperones/genetics/metabolism ; Motor Cortex/metabolism/pathology ; Motor Neurons/metabolism ; *Organ Specificity/genetics ; Protein Disulfide-Isomerases/genetics/metabolism ; Protein Interaction Maps/genetics ; *Proteostasis ; RNA, Messenger/genetics/metabolism ; *Signal Transduction ; Spinal Cord/metabolism/pathology ; *Unfolded Protein Response/genetics ; Vesicular Transport Proteins/metabolism ; Young Adult ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a disorder that affects motor neurons in motor cortex and spinal cord, and the degeneration of both neuronal populations is a critical feature of the disease. Abnormalities in protein homeostasis (proteostasis) are well established in ALS. However, they have been investigated mostly in spinal cord but less so in motor cortex. Herein, we monitored the unfolded protein (UPR) and heat shock response (HSR), two major proteostasis regulatory pathways, in human post-mortem tissue derived from the motor cortex of sporadic ALS (SALS) and compared them to those occurring in spinal cord. Although the UPR was activated in both tissues, specific expression of select UPR target genes, such as PDIs, was observed in motor cortex of SALS cases strongly correlating with oligodendrocyte markers. Moreover, we found that endoplasmic reticulum-associated degradation (ERAD) and HSR genes, which were activated predominately in spinal cord, correlated with the expression of neuronal markers. Our results indicate that proteostasis is strongly and selectively activated in SALS motor cortex and spinal cord where subsets of these genes are associated with specific cell type. This study expands our understanding of convergent molecular mechanisms occurring in motor cortex and spinal cord and highlights cell type-specific contributions.}, }
@article {pmid32334137, year = {2020}, author = {Katz, M and Davis, M and Garton, FC and Henderson, R and Bharti, V and Wray, N and McCombe, P}, title = {Mutations in heat shock protein beta-1 (HSPB1) are associated with a range of clinical phenotypes related to different patterns of motor neuron dysfunction: A case series.}, journal = {Journal of the neurological sciences}, volume = {413}, number = {}, pages = {116809}, doi = {10.1016/j.jns.2020.116809}, pmid = {32334137}, issn = {1878-5883}, mesh = {*Charcot-Marie-Tooth Disease/genetics ; *HSP27 Heat-Shock Proteins/genetics ; Heat-Shock Proteins/genetics ; Humans ; Molecular Chaperones ; Motor Neurons ; Mutation/genetics ; Phenotype ; }, abstract = {BACKGROUND: Heat shock protein beta-1 (HSPB1) is a ubiquitously expressed molecular chaperone that is important in protecting cells against cellular injury. Mutations in this protein are known to cause autosomal dominant hereditary distal axonal neuropathies, including Charcot Marie Tooth disease type 2F (CMT2F) and distal hereditary motor neuropathy (dHMN). However, patients with HSPB1 mutations have also been described with upper motor neuron signs. We present five patients with mutations in HSPB1 who presented with a range of clinical phenotypes related to different patterns of motor neuron dysfunction. Three of these mutations have not been previously reported.<br><br>METHODS: Patients were seen at our neuromuscular or amyotrophic lateral sclerosis (ALS) clinics. Gene sequencing was carried out as part of diagnostic investigations. Detailed clinical and electrophysiologic data was collected.<br><br>RESULTS: Five patients had variants of HSPB1. Three patients had a hereditary length-dependent sensori-motor axonal neuropathy consistent with Charcot Marie Tooth type 2 (CMT2); two of these patients carried novel mutations in the C-terminal region (p.Glu186* and p.Pro170Thr). One patient had the clinical picture of ALS and a novel missense mutation (p.Arg27Leu) in the N-terminal region. Another patient had the phenotype of hereditary spastic paraparesis (HSP) associated with a missense mutation (p.Gly84Arg) already described in families with CMT or dHMN.<br><br>CONCLUSION: This study describes three novel mutations of HSPB1 and describes two patients with upper motor neurone signs associated with HSPB1 mutations.}, }
@article {pmid32343854, year = {2020}, author = {Besnard-Guérin, C}, title = {Cytoplasmic localization of amyotrophic lateral sclerosis-related TDP-43 proteins modulates stress granule formation.}, journal = {The European journal of neuroscience}, volume = {52}, number = {8}, pages = {3995-4008}, doi = {10.1111/ejn.14762}, pmid = {32343854}, issn = {1460-9568}, mesh = {*Amyotrophic Lateral Sclerosis/genetics ; Cell Nucleus ; Cytoplasm ; DNA-Binding Proteins/genetics ; HeLa Cells ; Humans ; }, abstract = {TDP-43 is an RNA/DNA-binding protein associated with amyotrophic lateral sclerosis (ALS). Under pathological conditions, TDP-43 exported from the nucleus accumulates in the cytoplasm, forming inclusion bodies. However, the molecular mechanisms that contribute to such aggregation are unclear. The pathogenic processes that lead to aggregation in ALS were investigated by analysing the effects of wildtype human TDP-43 or with mutations in the nuclear localization sequence (NLS) or those associated with ALS in stress granule formation. TDP-43 (WT, ∆NLS or G348C), with or without a GFP-tag, was expressed in SH-SY5Y neuroblastoma or HeLa cells and stress granules induced by oxidative stress or heat shock. Stress granule formation was altered in cells strongly expressing GFP-TDP-∆NLS, or untagged TDP-43-∆NLS in the cytoplasm but not the negative controls, GFP or GFP-UtrCH. In contrast, there was no reduction in stress granule formation by cells that expressed untagged TDP-43 (WT or G348C) in the nucleus upon stress induction. GFP labelling of TDP-43 (WT or G348C) promotes high cytoplasmic expression and nuclear aggregation. Stress granule formation was impaired in cells expressing GFP-TDP-43 (WT or G348C) in the cytoplasm. Overall, these results suggest that stress granule formation may be inhibited by high levels of TDP-43 protein in the cytoplasm. As stress granules serve a protective function, their deregulation may promote neurodegeneration due to an aberrant stress response.}, }
@article {pmid32422904, year = {2020}, author = {Morimoto, S and Ishikawa, M and Watanabe, H and Isoda, M and Takao, M and Nakamura, S and Ozawa, F and Hirokawa, Y and Kuzuhara, S and Okano, H and Kokubo, Y}, title = {Brain Transcriptome Analysis Links Deficiencies of Stress-Responsive Proteins to the Pathomechanism of Kii ALS/PDC.}, journal = {Antioxidants (Basel, Switzerland)}, volume = {9}, number = {5}, pages = {}, pmid = {32422904}, issn = {2076-3921}, support = {17ek0109139h0003//Japan Agency for Medical Research and Development/ ; 19bm0804003h0003//Japan Agency for Medical Research and Development/ ; 25305030//Ministry of Education, Culture, Sports, Science and Technology/ ; 18KK0239//Ministry of Education, Culture, Sports, Science and Technology/ ; 17H01689//Ministry of Education, Culture, Sports, Science and Technology/ ; 18K07368//Ministry of Education, Culture, Sports, Science and Technology/ ; 15J03921//Ministry of Education, Culture, Sports, Science and Technology/ ; 19K17002//Ministry of Education, Culture, Sports, Science and Technology/ ; 15K09634//Ministry of Education, Culture, Sports, Science and Technology/ ; 18K07514//Ministry of Education, Culture, Sports, Science and Technology/ ; 21210301//Ministry of Health, Labour and Welfare/ ; 16H06277//Japan Society for the Promotion of Science/ ; H29-Nanchi- Ippan-085//the Research Committee of CNS Degenerative Diseases/ ; none//Japan Foundation for Neuroscience and Mental Health/ ; none//Japan Intractable Diseases (Nanbyo) Research Foundation/ ; none//the Mie Medical Fund/ ; }, abstract = {Amyotrophic lateral sclerosis and Parkinsonism-dementia complex (ALS/PDC) is a unique endemic neurodegenerative disease, with high-incidence foci in Kii Peninsula, Japan. To gather new insights into the pathological mechanisms underlying Kii ALS/PDC, we performed transcriptome analyses of patient brains. We prepared frozen brains from three individuals without neurodegenerative diseases, three patients with Alzheimer's disease, and 21 patients with Kii ALS/PDC, and then acquired microarray data from cerebral gray and white matter tissues. Microarray results revealed that expression levels of genes associated with heat shock proteins, DNA binding/damage, and senescence were significantly altered in patients with ALS/PDC compared with healthy individuals. The RNA expression pattern observed for ALS-type brains was similar to that of PDC-type brains. Additionally, pathway and network analyses indicated that the molecular mechanism underlying ALS/PDC may be associated with oxidative phosphorylation of mitochondria, ribosomes, and the synaptic vesicle cycle; in particular, upstream regulators of these mechanisms may be found in synapses and during synaptic trafficking. Furthermore, phenotypic differences between ALS-type and PDC-type were observed, based on HLA haplotypes. In conclusion, determining the relationship between stress-responsive proteins, synaptic dysfunction, and the pathogenesis of ALS/PDC in the Kii peninsula may provide new understanding of this mysterious disease.}, }
@article {pmid32531261, year = {2020}, author = {Shen, L and Wang, C and Chen, L and Leung, KL and Lo, E and Lakso, M and Wong, G}, title = {TDP-1/TDP-43 potentiates human α-Synuclein (HASN) neurodegeneration in Caenorhabditis elegans.}, journal = {Biochimica et biophysica acta. Molecular basis of disease}, volume = {1866}, number = {10}, pages = {165876}, doi = {10.1016/j.bbadis.2020.165876}, pmid = {32531261}, issn = {1879-260X}, mesh = {Animals ; Animals, Genetically Modified ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans Proteins/*genetics ; DNA-Binding Proteins/*genetics ; Disease Models, Animal ; Dopaminergic Neurons/pathology ; Gene Knockout Techniques ; Humans ; Lewy Body Disease/*genetics/pathology/physiopathology ; Locomotion/genetics ; Longevity/genetics ; Protein Aggregation, Pathological/*genetics/pathology/physiopathology ; RNA-Binding Proteins/*genetics ; alpha-Synuclein/*genetics/metabolism ; }, abstract = {TAR DNA binding protein (TDP-43) is a DNA/RNA binding protein whose pathological role in amyotrophic lateral sclerosis (ALS) and frontal temporal lobe dementia (FTLD) via formation of protein aggregates is well established. In contrast, knowledge concerning its interactions with other neuropathological aggregating proteins is poorly understood. Human α-synuclein (HASN) elicits dopaminergic neuron degeneration via protein aggregation in Parkinson's disease. HASN protein aggregates are also found in TDP-43 lesions and colocalize in Lewy Body Dementia (LBD). To better understand the interactions of TDP-43 and HASN, we investigated the effects of genetic deletion of tdp-1, the Caenorhabditis elegans ortholog of human TDP-43, as well as overexpression of TDP-43, in transgenic models overexpressing HASN[WT] and HASN[A53T]. Tdp-1 deletion improved the posture, movement, and developmental delay observed in transgenic animals pan-neuronally overexpressing HASN[A53T], and attenuated the loss and impairment of dopaminergic neurons caused by HASN[A53T] or HASN[WT] overexpression. Tdp-1 deletion also led to a decrease in protein level, mRNA level and aggregate formation of HASN in living animals. RNA-seq studies suggested that tdp-1 supports expression of lysosomal genes and decreases expression of genes involved in heat shock. RNAi demonstrated that heat shock proteins can mediate HASN neuropathology. Co-overexpression of both human TDP-43 and HASN[WT] resulted in locomotion deficits, shorter lifespan, and more severe dopaminergic neuron impairments compared to single transgenes. Our results suggest TDP-1/TDP-43 potentiates HASN mediated neurodegeneration in C. elegans. This study indicates a multifunctional role for TDP-1/TDP-43 in neurodegeneration involving HASN.}, }
@article {pmid32535145, year = {2020}, author = {Melnik, A and Cappelletti, V and Vaggi, F and Piazza, I and Tognetti, M and Schwarz, C and Cereghetti, G and Ahmed, MA and Soste, M and Matlack, K and de Souza, N and Csikasz-Nagy, A and Picotti, P}, title = {Comparative analysis of the intracellular responses to disease-related aggregation-prone proteins.}, journal = {Journal of proteomics}, volume = {225}, number = {}, pages = {103862}, doi = {10.1016/j.jprot.2020.103862}, pmid = {32535145}, issn = {1876-7737}, mesh = {Humans ; *Neurodegenerative Diseases ; Proteome ; *Proteomics ; }, abstract = {Aggregation-prone proteins (APPs) have been implicated in numerous human diseases but the underlying mechanisms are incompletely understood. Here we comparatively analysed cellular responses to different APPs. Our study is based on a systematic proteomic and phosphoproteomic analysis of a set of yeast proteotoxicity models expressing different human disease-related APPs, which accumulate intracellular APP inclusions and exhibit impaired growth. Clustering and functional enrichment analyses of quantitative proteome-level data reveal that the cellular response to APP expression, including the chaperone response, is specific to the APP, and largely differs from the response to a more generalized proteotoxic insult such as heat shock. We further observe an intriguing association between the subcellular location of inclusions and the location of the cellular response, and provide a rich dataset for future mechanistic studies. Our data suggest that care should be taken when designing research models to study intracellular aggregation, since the cellular response depends markedly on the specific APP and the location of inclusions. Further, therapeutic approaches aimed at boosting protein quality control in protein aggregation diseases should be tailored to the subcellular location affected by inclusion formation. SIGNIFICANCE: We have examined the global cellular response, in terms of protein abundance and phosphorylation changes, to the expression of five human neurodegeneration-associated, aggregation-prone proteins (APPs) in a set of isogenic yeast models. Our results show that the cellular response to each APP is unique to that protein, is different from the response to thermal stress, and is associated with processes at the subcellular location of APP inclusion formation. These results further our understanding of how cells, in a model organism, respond to expression of APPs implicated in neurodegenerative diseases like Parkinson's, Alzheimer's, and ALS. They have implications for mechanisms of toxicity as well as of protective responses in the cell. The specificity of the response to each APP means that research models of these diseases should be tailored to the APP in question. The subcellular localization of the response suggest that therapeutic interventions should also be targeted within the cell.}, }
@article {pmid32661089, year = {2020}, author = {San Gil, R and Cox, D and McAlary, L and Berg, T and Walker, AK and Yerbury, JJ and Ooi, L and Ecroyd, H}, title = {Neurodegenerative disease-associated protein aggregates are poor inducers of the heat shock response in neuronal cells.}, journal = {Journal of cell science}, volume = {133}, number = {15}, pages = {}, doi = {10.1242/jcs.243709}, pmid = {32661089}, issn = {1477-9137}, mesh = {Heat Shock Transcription Factors/genetics ; Heat-Shock Response/genetics ; Humans ; *Neurodegenerative Diseases/genetics ; Protein Aggregates ; Superoxide Dismutase-1 ; }, abstract = {Protein aggregates that result in inclusion formation are a pathological hallmark common to many neurodegenerative diseases, including amyotrophic lateral sclerosis, Parkinson's disease and Huntington's disease. Under conditions of cellular stress, activation of the heat shock response (HSR) results in an increase in the levels of molecular chaperones and is a first line of cellular defence against inclusion formation. It remains to be established whether neurodegenerative disease-associated proteins and inclusions are themselves capable of inducing an HSR in neuronal cells. To address this, we generated a neuroblastoma cell line that expresses a fluorescent reporter protein under conditions of heat shock transcription factor 1 (HSF1)-mediated HSR induction. We show that the HSR is not induced by exogenous treatment with aggregated forms of recombinant α-synuclein or the G93A mutant of superoxide dismutase-1 (SOD1[G93A]) nor intracellular expression of SOD1[G93A] or a pathogenic form of polyglutamine-expanded huntingtin (Htt[72Q]). These results suggest that pathogenic proteins evade detection or impair induction of the HSR in neuronal cells. A failure of protein aggregation to induce an HSR might contribute to the development of inclusion pathology in neurodegenerative diseases.This article has an associated First Person interview with the first author of the paper.}, }
@article {pmid32686212, year = {2020}, author = {Watabe, K and Kato, Y and Sakuma, M and Murata, M and Niida-Kawaguchi, M and Takemura, T and Hanagata, N and Tada, M and Kakita, A and Shibata, N}, title = {Praja1 RING-finger E3 ubiquitin ligase suppresses neuronal cytoplasmic TDP-43 aggregate formation.}, journal = {Neuropathology : official journal of the Japanese Society of Neuropathology}, volume = {40}, number = {6}, pages = {570-586}, pmid = {32686212}, issn = {1440-1789}, support = {//Collaborative Research Project (2017-2019) of Brain Research Institute, Niigata University/ ; 15K06763, 18K06507//Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (JSPS KAKENHI)/ ; //Niigata University/ ; //Ministry of Education/ ; }, mesh = {Amyotrophic Lateral Sclerosis/metabolism/pathology ; Animals ; Cytoplasm/pathology ; DNA-Binding Proteins/*metabolism ; Heat Shock Transcription Factors/metabolism ; Humans ; Mice ; Neurons/*pathology ; Protein Aggregation, Pathological/*metabolism ; Rats ; TDP-43 Proteinopathies/metabolism/pathology ; Ubiquitin-Protein Ligases/*metabolism ; }, abstract = {Transactivation response DNA-binding protein of 43 kDa (TDP-43) is a major constituent of cytoplasmic aggregates in neuronal and glial cells in cases of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We have previously shown neuronal cytoplasmic aggregate formation induced by recombinant adenoviruses expressing human wild-type and C-terminal fragment (CTF) TDP-43 under the condition of proteasome inhibition in vitro and in vivo. In the present study, we demonstrated that the formation of the adenoviral TDP-43 aggregates was markedly suppressed in rat neural stem cell-derived neuronal cells by co-infection of an adenovirus expressing heat shock transcription factor 1 (HSF1), a master regulator of heat shock response. We performed DNA microarray analysis and searched several candidate molecules, located downstream of HSF1, which counteract TDP-43 aggregate formation. Among these, we identified Praja 1 RING-finger E3 ubiquitin ligase (PJA1) as a suppressor of phosphorylation and aggregate formation of TDP-43. Co-immunoprecipitation assay revealed that PJA1 binds to CTF TDP-43 and the E2-conjugating enzyme UBE2E3. PJA1 also suppressed formation of cytoplasmic phosphorylated TDP-43 aggregates in mouse facial motor neurons in vivo. Furthermore, phosphorylated TDP-43 aggregates were detected in PJA1-immunoreactive human ALS motor neurons. These results indicate that PJA1 is one of the principal E3 ubiquitin ligases for TDP-43 to counteract its aggregation propensity and could be a potential therapeutic target for ALS and FTLD.}, }
@article {pmid32687490, year = {2020}, author = {Ryu, SW and Stewart, R and Pectol, DC and Ender, NA and Wimalarathne, O and Lee, JH and Zanini, CP and Harvey, A and Huibregtse, JM and Mueller, P and Paull, TT}, title = {Proteome-wide identification of HSP70/HSC70 chaperone clients in human cells.}, journal = {PLoS biology}, volume = {18}, number = {7}, pages = {e3000606}, pmid = {32687490}, issn = {1545-7885}, support = {/HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Cell Line ; HSC70 Heat-Shock Proteins/*metabolism ; HSP70 Heat-Shock Proteins/*metabolism ; Humans ; Mutation/genetics ; Protein Binding ; Protein Biosynthesis ; Protein Folding ; Proteome/*metabolism ; Substrate Specificity ; Ubiquitin/metabolism ; }, abstract = {The 70 kDa heat shock protein (HSP70) family of chaperones are the front line of protection from stress-induced misfolding and aggregation of polypeptides in most organisms and are responsible for promoting the stability, folding, and degradation of clients to maintain cellular protein homeostasis. Here, we demonstrate quantitative identification of HSP70 and 71 kDa heat shock cognate (HSC70) clients using a ubiquitin-mediated proximity tagging strategy and show that, despite their high degree of similarity, these enzymes have largely nonoverlapping specificities. Both proteins show a preference for association with newly synthesized polypeptides, but each responds differently to changes in the stoichiometry of proteins in obligate multi-subunit complexes. In addition, expression of an amyotrophic lateral sclerosis (ALS)-associated superoxide dismutase 1 (SOD1) mutant protein induces changes in HSP70 and HSC70 client association and aggregation toward polypeptides with predicted disorder, indicating that there are global effects from a single misfolded protein that extend to many clients within chaperone networks. Together these findings show that the ubiquitin-activated interaction trap (UBAIT) fusion system can efficiently isolate the complex interactome of HSP chaperone family proteins under normal and stress conditions.}, }
@article {pmid32726594, year = {2021}, author = {Une, M and Yamakawa, M and Watanabe, Y and Uchino, K and Honda, N and Adachi, M and Nakanishi, M and Umezawa, A and Kawata, Y and Nakashima, K and Hanajima, R}, title = {SOD1-interacting proteins: Roles of aggregation cores and protein degradation systems.}, journal = {Neuroscience research}, volume = {170}, number = {}, pages = {295-305}, doi = {10.1016/j.neures.2020.07.010}, pmid = {32726594}, issn = {1872-8111}, mesh = {*Amyotrophic Lateral Sclerosis ; Animals ; Disease Models, Animal ; Mice ; Mice, Transgenic ; Motor Neurons/metabolism ; Mutation ; Proteolysis ; Spinal Cord/metabolism ; *Superoxide Dismutase/metabolism ; Superoxide Dismutase-1/genetics/metabolism ; }, abstract = {Cu/Zn superoxide dismutase (SOD1) mutations are associated with amyotrophic lateral sclerosis (ALS). SOD1-positive aggregates in motor neurons, as well as proteins that interact with the aggregates are presumably involved in ALS neurotoxicity. We used a proteomics approach to compare differences in protein expression in spinal cord homogenates from non-transgenic (NTG) and ALS model mice. Using the homogenates, we identified proteins that interacted with SOD1 seeds in vitro. We assessed differences in SOD1-interacting proteins in cell cultures treated with proteasome or autophagy inhibitor. In the first experiment, intermediate filamentous and small heat shock proteins were upregulated in glial cells. We identified 26 protein types that interacted with aggregation cores in ALS model homogenates, and unexpectedly, 40 proteins in were detected in NTG mice. In cell cultures treated with proteasome and autophagy inhibitors, we identified 16 and 11 SOD1-interacting proteins, respectively, and seven proteins in untreated cells. These SOD1-interacting proteins were involved in multiple cellular functions such as protein quality control, cytoskeletal organization, and pathways involved in growth factor signaling and their downstream cascades. The complex interactions between pathways could cause further dysregulation, ultimately leading to fatal cellular dysfunction in ALS.}, }
@article {pmid32794552, year = {2020}, author = {Wright, GSA}, title = {Molecular and pharmacological chaperones for SOD1.}, journal = {Biochemical Society transactions}, volume = {48}, number = {4}, pages = {1795-1806}, pmid = {32794552}, issn = {1470-8752}, mesh = {Animals ; Copper/metabolism ; Heat-Shock Proteins/metabolism ; Humans ; Macrophage Migration-Inhibitory Factors/metabolism ; Molecular Chaperones/*metabolism ; Mutation ; Protein Folding ; Superoxide Dismutase-1/genetics/*metabolism ; }, abstract = {The efficacy of superoxide dismutase-1 (SOD1) folding impacts neuronal loss in motor system neurodegenerative diseases. Mutations can prevent SOD1 post-translational processing leading to misfolding and cytoplasmic aggregation in familial amyotrophic lateral sclerosis (ALS). Evidence of immature, wild-type SOD1 misfolding has also been observed in sporadic ALS, non-SOD1 familial ALS and Parkinson's disease. The copper chaperone for SOD1 (hCCS) is a dedicated and specific chaperone that assists SOD1 folding and maturation to produce the active enzyme. Misfolded or misfolding prone SOD1 also interacts with heat shock proteins and macrophage migration inhibitory factor to aid folding, refolding or degradation. Recognition of specific SOD1 structures by the molecular chaperone network and timely dissociation of SOD1-chaperone complexes are, therefore, important steps in SOD1 processing. Harnessing these interactions for therapeutic benefit is actively pursued as is the modulation of SOD1 behaviour with pharmacological and peptide chaperones. This review highlights the structural and mechanistic aspects of a selection of SOD1-chaperone interactions together with their impact on disease models.}, }
@article {pmid32929655, year = {2020}, author = {Nagata, E and Fujii, N and Kohara, S and Okada, C and Satoh, T and Takekoshi, S and Takao, M and Mihara, B and Takizawa, S}, title = {Inositol hexakisphosphate kinase 2 promotes cell death of anterior horn cells in the spinal cord of patients with amyotrophic lateral sclerosis.}, journal = {Molecular biology reports}, volume = {47}, number = {9}, pages = {6479-6485}, pmid = {32929655}, issn = {1573-4978}, support = {JP 19K07851//Japan Society for the Promotion of Science/ ; 16H06277//Japan Society for the Promotion of Science/ ; }, mesh = {Aged ; Aged, 80 and over ; Amyotrophic Lateral Sclerosis/enzymology/genetics/*metabolism ; Anterior Horn Cells/enzymology/*metabolism ; Autopsy ; Casein Kinase II/genetics/metabolism ; Cell Death/*genetics ; Cell Nucleus/genetics/metabolism ; Cytoplasm/genetics/metabolism ; Female ; Gene Expression Regulation/genetics ; HSP90 Heat-Shock Proteins/genetics/metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Phosphorylation ; Phosphotransferases (Phosphate Group Acceptor)/genetics/*metabolism ; Pleckstrin Homology Domains ; Protein Domains ; Proto-Oncogene Proteins c-akt/genetics/metabolism ; Spinal Cord/cytology/*metabolism/pathology ; }, abstract = {We have previously reported that inositol hexakisphosphate kinase (InsP6K)2 mediates cell death. InsP6K2 is abundantly expressed in anterior horn cells of the mammalian spinal cord. We investigated the role of InsP6K2 in spinal cords of patients with amyotrophic lateral sclerosis (ALS). Autopsy specimens of lumbar spinal cords from ten patients with sporadic ALS and five non-neurological disease patients (NNDPs) were obtained. We performed quantitative real-time PCR, immunostaining, and western blotting for InsP6K1, InsP6K2, InsP6K3, protein kinase B (Akt), casein kinase 2 (CK2), and 90-kDa heat-shock protein (HSP90). In contrast to InsP6K1 and InsP6K3 mRNA expression, InsP6K2 levels in anterior horn cells of the spinal cord were significantly increased in ALS patients compared to NNDPs. In ALS patients, InsP6K2 translocated from the nucleus to the cytoplasm. However, we observed a decrease in HSP90, CK2, and Akt activity in ALS patients compared to NNDPs. A previous study reported that InsP6K2 activity is suppressed after binding to HSP90 and subsequent phosphorylation and degradation by CK2, thus decreasing InsP6K2 activity. However, InsP7, which is generated by InsP6K2, can compete with Akt for PH domain binding. Consequently, InsP7 can inhibit Akt phosphorylation. Our results suggest that InsP6K2 is activated in the spinal cord of patients with ALS and may play an important role in ALS by inducing cell death mechanisms via Akt, CK2, and HSP90 pathways.}, }
@article {pmid33193563, year = {2020}, author = {Wang, M and Liu, Z and Yuan, Y and Ni, J and Li, W and Hu, Y and Liu, P and Hou, X and Huang, L and Jiao, B and Shen, L and Jiang, H and Tang, B and Wang, J}, title = {A Novel Potentially Pathogenic Rare Variant in the DNAJC7 Gene Identified in Amyotrophic Lateral Sclerosis Patients From Mainland China.}, journal = {Frontiers in genetics}, volume = {11}, number = {}, pages = {821}, pmid = {33193563}, issn = {1664-8021}, abstract = {Variants in the DNAJC7 gene have been shown to be novel causes of amyotrophic lateral sclerosis (ALS). However, the contributions of DNAJC7 mutations in Asian ALS patients remain unclear. In this study, we screened rare pathogenic variants in the DNAJC7 gene in a cohort of 578 ALS patients from Mainland China. A novel, rare, putative pathogenic variant c.712A>G (p.R238G) was identified in one sporadic ALS patient. The carrier with this variant exhibited symptom onset at a relatively younger age and experienced rapid disease progression. Our results expand the pathogenic variant spectrum of DNAJC7 and indicate that variants in the DNAJC7 gene may also contribute to ALS in the Chinese population.}, }
@article {pmid33557211, year = {2021}, author = {Martin, LJ and Niedzwiecki, MV and Wong, M}, title = {Chronic Intermittent Mild Whole-Body Hypothermia Is Therapeutic in a Mouse Model of ALS.}, journal = {Cells}, volume = {10}, number = {2}, pages = {}, pmid = {33557211}, issn = {2073-4409}, support = {NS34100/NH/NIH HHS/United States ; }, mesh = {Acclimatization ; Amyotrophic Lateral Sclerosis/complications/*therapy ; Animals ; Chronic Disease ; Cold Temperature ; Cytoprotection ; Disease Models, Animal ; Fever/complications ; Humans ; *Hypothermia, Induced ; Inflammation/pathology ; Male ; Mice, Transgenic ; Mitochondria, Muscle/metabolism ; Mitochondrial Permeability Transition Pore/metabolism ; Motor Neurons/pathology ; Muscle, Skeletal/metabolism/pathology ; Neuromuscular Junction/pathology ; Spinal Cord/pathology ; Superoxide Dismutase-1/metabolism ; Survival Analysis ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that causes motor neuron degeneration. There are no cures or effective treatments for ALS. Therapeutic hypothermia is effectively used clinically to mitigate mortality in patients with acute acquired brain injury and in surgical settings to minimize secondary brain injury. The efficacy of therapeutic hypothermia in chronic neurodegenerative disorders has not been examined. We tested the hypothesis that mild hypothermia/cold acclimation is therapeutic in a transgenic mouse model of ALS caused by expression of mutated human superoxide dismutase-1 gene. At presymptomatic stages of disease, body temperatures (oral and axial) of mutant male mice were persistently hyperthermic (38-38.5 °C) compared to littermate controls, but at end-stage disease mice were generally hypothermic (36-36.5 °C). Presymptomatic mutant mice (awake-freely moving) were acclimated to systemic mild hypothermia using an environmentally controlled chamber (12 h-on/12-off or 24 h-on/24 h-off) to lower body temperature (1-3 °C). Cooled ALS mice showed a significant delay in disease onset (103-112 days) compared to normothermia mice (80-90 days) and exhibited significant attenuation of functional decline in motor performance. Cooled mice examined at 80 days had reduced motor neuron loss, mitochondrial swelling, and spinal cord inflammation compared to non-cooled mice. Cooling attenuated the loss of heat-shock protein 70, mitochondrial uncoupling protein-3, and sumoylated-1 (SUMO1)-conjugated proteins in skeletal muscle and disengaged the mitochondrial permeability transition pore. Cooled ALS mice had a significant extension of lifespan (148 ± 7 days) compared to normothermic mice (135 ± 4 days). Thus, intermittent systemic mild hypothermia is therapeutic in mouse ALS with protective effects manifested within the CNS and skeletal muscle that target mitochondria.}, }
@article {pmid33705388, year = {2021}, author = {Lu, YN and Kavianpour, S and Zhang, T and Zhang, X and Nguyen, D and Thombre, R and He, L and Wang, J}, title = {MARK2 phosphorylates eIF2α in response to proteotoxic stress.}, journal = {PLoS biology}, volume = {19}, number = {3}, pages = {e3001096}, pmid = {33705388}, issn = {1545-7885}, support = {R01 NS074324/NS/NINDS NIH HHS/United States ; R01 NS089616/NS/NINDS NIH HHS/United States ; R01 NS110098/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Cell Line ; Disease Models, Animal ; Endoplasmic Reticulum/metabolism ; Eukaryotic Initiation Factor-2/*metabolism/physiology ; HSP90 Heat-Shock Proteins/metabolism ; Homeostasis ; Humans ; Mice ; Mice, Knockout ; Microtubules/metabolism ; Phosphorylation ; Protein Biosynthesis ; Protein Serine-Threonine Kinases/*metabolism ; Stress, Physiological/physiology ; eIF-2 Kinase/metabolism ; }, abstract = {The regulation of protein synthesis is essential for maintaining cellular homeostasis, especially during stress responses, and its dysregulation could underlie the development of human diseases. The critical step during translation regulation is the phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α). Here we report the identification of a direct kinase of eIF2α, microtubule affinity-regulating kinase 2 (MARK2), which phosphorylates eIF2α in response to proteotoxic stress. The activity of MARK2 was confirmed in the cells lacking the 4 previously known eIF2α kinases. MARK2 itself was found to be a substrate of protein kinase C delta (PKCδ), which serves as a sensor for protein misfolding stress through a dynamic interaction with heat shock protein 90 (HSP90). Both MARK2 and PKCδ are activated via phosphorylation in proteotoxicity-associated neurodegenerative mouse models and in human patients with amyotrophic lateral sclerosis (ALS). These results reveal a PKCδ-MARK2-eIF2α cascade that may play a critical role in cellular proteotoxic stress responses and human diseases.}, }
@article {pmid33749723, year = {2021}, author = {Straub, IR and Weraarpachai, W and Shoubridge, EA}, title = {Multi-OMICS study of a CHCHD10 variant causing ALS demonstrates metabolic rewiring and activation of endoplasmic reticulum and mitochondrial unfolded protein responses.}, journal = {Human molecular genetics}, volume = {30}, number = {8}, pages = {687-705}, pmid = {33749723}, issn = {1460-2083}, support = {//CIHR/Canada ; }, mesh = {Amyotrophic Lateral Sclerosis/*genetics/metabolism ; Cells, Cultured ; Endoplasmic Reticulum/genetics/metabolism ; Endoribonucleases/genetics/metabolism ; Gene Expression Profiling/*methods ; Gene Ontology ; Humans ; Metabolic Networks and Pathways/genetics ; Metabolomics/*methods ; Mitochondria/genetics/metabolism ; Mitochondrial Proteins/*genetics/metabolism ; *Mutation ; Protein Serine-Threonine Kinases/genetics/metabolism ; Proteome/metabolism ; Proteomics/*methods ; Signal Transduction/genetics ; Unfolded Protein Response/genetics ; X-Box Binding Protein 1/genetics/metabolism ; }, abstract = {Mutations in CHCHD10, coding for a mitochondrial intermembrane space protein, are a rare cause of autosomal dominant amyotrophic lateral sclerosis. Mutation-specific toxic gain of function or haploinsufficiency models have been proposed to explain pathogenicity. To decipher the metabolic dysfunction associated with the haploinsufficient p.R15L variant, we integrated transcriptomic, metabolomic and proteomic data sets in patient cells subjected to an energetic stress that forces the cells to rely on oxidative phosphorylation for ATP production. Patient cells had a complex I deficiency that resulted in an increased NADH/NAD+ ratio, diminished TCA cycle activity, a reorganization of one carbon metabolism and an increased AMP/ATP ratio leading to phosphorylation of AMPK and inhibition of mTORC1. These metabolic changes activated the unfolded protein response (UPR) in the ER through the IRE1/XBP1 pathway, upregulating downstream targets including ATF3, ATF4, CHOP and EGLN3, and two cytokine markers of mitochondrial disease, GDF15 and FGF21. Activation of the mitochondrial UPR was mediated through an upregulation of the transcription factors ATF4 and ATF5, leading to increased expression of mitochondrial proteases and heat shock proteins. There was a striking transcriptional up regulation of at least seven dual specific phosphatases, associated with an almost complete dephosphorylation of JNK isoforms, suggesting a concerted deactivation of MAP kinase pathways. This study demonstrates that loss of CHCHD10 function elicits an energy deficit that activates unique responses to nutrient stress in both the mitochondria and ER, which may contribute to the selective vulnerability of motor neurons.}, }
@article {pmid34068231, year = {2021}, author = {Malcova, I and Senohrabkova, L and Novakova, L and Hasek, J}, title = {eIF3a Destabilization and TDP-43 Alter Dynamics of Heat-Induced Stress Granules.}, journal = {International journal of molecular sciences}, volume = {22}, number = {10}, pages = {}, pmid = {34068231}, issn = {1422-0067}, support = {GACU1180213//Grantov&#xe1; Agentura, Univerzita Karlova/ ; CSF16-05497S//Czech Science Foundation/ ; }, mesh = {Cytoplasmic Granules/*physiology ; DNA-Binding Proteins/genetics/*metabolism ; Endoplasmic Reticulum/metabolism ; Eukaryotic Initiation Factor-3/*chemistry/genetics/metabolism ; *Heat-Shock Response ; Humans ; Mitochondria/metabolism ; Protein Stability ; RNA, Messenger/genetics/metabolism ; Saccharomyces cerevisiae/genetics/growth &amp; development/*metabolism ; Saccharomyces cerevisiae Proteins/genetics/*metabolism ; }, abstract = {Stress granules (SGs) are membrane-less assemblies arising upon various stresses in eukaryotic cells. They sequester mRNAs and proteins from stressful conditions and modulate gene expression to enable cells to resume translation and growth after stress relief. SGs containing the translation initiation factor eIF3a/Rpg1 arise in yeast cells upon robust heat shock (HS) at 46 °C only. We demonstrate that the destabilization of Rpg1 within the PCI domain in the Rpg1-3 variant leads to SGs assembly already at moderate HS at 42 °C. These are bona fide SGs arising upon translation arrest containing mRNAs, which are components of the translation machinery, and associating with P-bodies. HS SGs associate with endoplasmatic reticulum and mitochondria and their contact sites ERMES. Although Rpg1-3-labeled SGs arise at a lower temperature, their disassembly is delayed after HS at 46 °C. Remarkably, the delayed disassembly of HS SGs after the robust HS is reversed by TDP-43, which is a human protein connected with amyotrophic lateral sclerosis. TDP-43 colocalizes with HS SGs in yeast cells and facilitates cell regrowth after the stress relief. Based on our results, we propose yeast HS SGs labeled by Rpg1 and its variants as a novel model system to study functions of TDP-43 in stress granules disassembly.}, }
@article {pmid34069691, year = {2021}, author = {San Gil, R and Clarke, BE and Ecroyd, H and Kalmar, B and Greensmith, L}, title = {Regional Differences in Heat Shock Protein 25 Expression in Brain and Spinal Cord Astrocytes of Wild-Type and SOD1 [G93A] Mice.}, journal = {Cells}, volume = {10}, number = {5}, pages = {}, pmid = {34069691}, issn = {2073-4409}, support = {G0601943//UK Medical Research Council Centre for Neuromuscular Diseases/ ; }, mesh = {Animals ; Astrocytes/drug effects/*enzymology/pathology ; Cells, Cultured ; Cerebral Cortex/drug effects/*enzymology/pathology ; Female ; Gene Expression Regulation ; Gliosis/enzymology/pathology ; Heat-Shock Proteins/genetics/*metabolism ; Heat-Shock Response ; Humans ; Lipopolysaccharides/pharmacology ; Male ; Mice, Inbred C57BL ; Mice, Transgenic ; Molecular Chaperones/genetics/*metabolism ; Phenotype ; Spinal Cord/drug effects/*enzymology/pathology ; Superoxide Dismutase-1/genetics/*metabolism ; Tumor Necrosis Factor-alpha/pharmacology ; Mice ; }, abstract = {Heterogeneity of glia in different CNS regions may contribute to the selective vulnerability of neuronal populations in neurodegenerative conditions such as amyotrophic lateral sclerosis (ALS). Here, we explored regional variations in the expression of heat shock protein 25 in glia under conditions of acute and chronic stress. Hsp27 (Hsp27; murine orthologue: Hsp25) fulfils a number of cytoprotective functions and may therefore be a possible therapeutic target in ALS. We identified a subpopulation of astrocytes in primary murine mixed glial cultures that expressed Hsp25. Under basal conditions, the proportion of Hsp25-positive astrocytes was twice as high in spinal cord cultures than in cortical cultures. To explore the physiological role of the elevated Hsp25 expression in spinal cord astrocytes, we exposed cortical and spinal cord glia to acute stress, using heat stress and pro-inflammatory stimuli. Surprisingly, we observed no stress-induced increase in Hsp25 expression in either cortical or spinal cord astrocytes. Similarly, exposure to endogenous stress, as modelled in glial cultures from SOD1 [G93A]-ALS mice, did not increase Hsp25 expression above that observed in astrocytes from wild-type mice. In vivo, Hsp25 expression was greater under conditions of chronic stress present in the spinal cord of SOD1 [G93A] mice than in wild-type mice, although this increase in expression is likely to be due to the extensive gliosis that occurs in this model. Together, these results show that there are differences in the expression of Hsp25 in astrocytes in different regions of the central nervous system, but Hsp25 expression is not upregulated under acute or chronic stress conditions.}, }
@article {pmid34085462, year = {2021}, author = {Liu, XW and Yan, Y and Hei, YF and Zhu, CJ and Tian, Q and Sun, WJ}, title = {[Effect of electroacupuncture on expression of heat shock protein 70 in mice with amyotrophic lateral sclerosis].}, journal = {Zhen ci yan jiu = Acupuncture research}, volume = {46}, number = {5}, pages = {391-396}, doi = {10.13702/j.1000-0607.200639}, pmid = {34085462}, issn = {1000-0607}, mesh = {*Amyotrophic Lateral Sclerosis/genetics/therapy ; Animals ; *Electroacupuncture ; HSP70 Heat-Shock Proteins/genetics ; Mice ; Mice, Inbred C57BL ; }, abstract = {OBJECTIVE: To observe the effects of electroacupuncture (EA) on the body weight, disease progression and the expression of heat shock protein 70 (HSP70) in lumbar spinal cord of amyotrophic lateral sclerosis (ALS) mice, so as to explore the mechanism of EA on treating ALS.<br><br>METHODS: Eighteen ALS transgenic SOD1[G93A] mice were randomly divided into model, EA and medication groups, with 6 mice in each group, and six C57BL/6J mice were used as the normal group. Mice in the EA group received EA at "Quchi"(LI11)-"Hegu"(LI4), "Zusanli"(ST36)- "Sanyinjiao"(SP6), 30 min/time, 5 times/week, for 8 weeks.Mice in the medication group were given riluzole solution (7.6 mg&#xb7;kg[-1]&#xb7;d[-1]) by gavage for 8 weeks. The body weight of the mice was recorded and the motor function of the hind limbs was evaluated by the neurological function scoring stan-dard of the ALS Therapeutic Development Institute. The expression of HSP70 in lumbar spinal cord was detected by Western blot and immunohistochemistry, respectively.<br><br>RESULTS: Compared with the normal group, the body weight and the expression of HSP70 in the model group decreased significantly (P<0.05), while no significant difference in the body weight was found among other groups(P>0.05). After intervention and in comparison with the model group, the disease onset time and paralysis time of the EA group and the medication group were significantly delayed (P<0.05, P<0.01), the expression of HSP70 in the EA group and the medicine group was significantly increased (P<0.05, P<0.01).But there was no significant difference in the survival time among all groups(P&#xff1e;0.05). The disease onset time of the EA group was shorter than that in the medication group (P<0.05).<br><br>CONCLUSION: EA can increase the expression of HSP70 in lumbar spinal cord, thereby delaying the progression of ALS.}, }
@article {pmid34258561, year = {2021}, author = {Kakihana, T and Takahashi, M and Katsuragi, Y and Yamashita, SI and Sango, J and Kanki, T and Onodera, O and Fujii, M}, title = {The optineurin/TIA1 pathway inhibits aberrant stress granule formation and reduces ubiquitinated TDP-43.}, journal = {iScience}, volume = {24}, number = {7}, pages = {102733}, pmid = {34258561}, issn = {2589-0042}, abstract = {Amyotrophic lateral sclerosis (ALS) is a degenerative motor neuron disease characterized by the formation of cytoplasmic ubiquitinated TDP-43 protein aggregates in motor neurons. Stress granules (SGs) are stress-induced cytoplasmic protein aggregates containing various neuropathogenic proteins, including TDP-43. Several studies have suggested that SGs are the initial site of the formation of pathogenic ubiquitinated TDP-43 aggregates in ALS neurons. Mutations in the optineurin (OPTN) and TIA1 genes are causative factors of familial ALS with TDP-43 aggregation pathology. We found that both OPTN depletion and ALS-associated OPTN mutations upregulated the TIA1 level in cells recovered from heat shock, and this upregulated TIA1 increased the amount of ubiquitinated TDP-43. Ubiquitinated TDP-43 induced by OPTN depletion was localized in SGs. Our study suggests that ALS-associated loss-of-function mutants of OPTN increase the amount of ubiquitinated TDP-43 in neurons by increasing the expression of TIA1, thereby promoting the aggregation of ubiquitinated TDP-43.}, }
@article {pmid34506621, year = {2021}, author = {Lim, SJ and Mohamad Ali, MS and Sabri, S and Muhd Noor, ND and Salleh, AB and Oslan, SN}, title = {Opportunistic yeast pathogen Candida spp.: Secreted and membrane-bound virulence factors.}, journal = {Medical mycology}, volume = {59}, number = {12}, pages = {1127-1144}, doi = {10.1093/mmy/myab053}, pmid = {34506621}, issn = {1460-2709}, support = {FRGS/1/2019/STG05/UPM/02/1//Ministry of Higher Education, Malaysia/ ; }, mesh = {Animals ; Antifungal Agents/therapeutic use ; *Candida ; Candida albicans ; *Candidiasis, Invasive/drug therapy/veterinary ; Virulence Factors ; }, abstract = {UNLABELLED: Candidiasis is a fungal infection caused by Candida spp. especially Candida albicans, C. glabrata, C. parapsilosis and C. tropicalis. Although the medicinal therapeutic strategies have rapidly improved, the mortality rate as candidiasis has continuously increased. The secreted and membrane-bound virulence factors (VFs) are responsible for fungal invasion, damage and translocation through the host enterocytes besides the evasion from host immune system. VFs such as agglutinin-like sequences (Als), heat shock protein 70, phospholipases, secreted aspartyl proteinases (Sap), lipases, enolases and phytases are mostly hydrolases which degrade or interact with the enterocyte membrane components. Candidalysin, however, acts as a peptide toxin to induce necrotic cell lysis. To date, structural studies of the VFs remain underexplored, hindering their functional analyses. Among the VFs, only Sap and Als have their structures deposited in Protein Data Bank (PDB). Therefore, this review scrutinizes the mechanisms of these VFs by discussing the VF-deficient studies of several Candida spp. and their abilities to produce these VFs. Nonetheless, their latest reported sequential and structural analyses are discussed to impart a wider perception of the host-pathogen interactions and potential vaccine or antifungal drug targets. This review signifies that more VFs structural investigations and mining in the emerging Candida spp. are required to decipher their pathogenicity and virulence mechanisms compared to the prominent C. albicans.<br><br>LAY SUMMARY: Candida virulence factors (VFs) including mainly enzymes and proteins play vital roles in breaching the human intestinal barrier and causing deadly invasive candidiasis. Limited VFs' structural studies hinder deeper comprehension of their mechanisms and thus the design of vaccines and antifungal drugs against fungal infections.}, }
@article {pmid34685574, year = {2021}, author = {Wang, L and Bergkvist, L and Kumar, R and Winblad, B and Pavlov, PF}, title = {Targeting Chaperone/Co-Chaperone Interactions with Small Molecules: A Novel Approach to Tackle Neurodegenerative Diseases.}, journal = {Cells}, volume = {10}, number = {10}, pages = {}, pmid = {34685574}, issn = {2073-4409}, support = {2018-02843//Swedish Research Council/ ; AF940014//Alzheimer Foundation/ ; Fo 2019-0140//Brain Foundation/ ; 2021-00681//Foundation for Geriatric Diseases at Karolinska Institutet/ ; 0//Gunvor and Josef An&#xe9;rs Foundation/ ; 0//Gun and Bertil Stohnes Foundation/ ; 0//Tore Nilssons Foundation for medical research/ ; 0//Mar-garetha af Ugglas Foundation/ ; 2020-01028//the Foundation for Old Servants/ ; 0//Magnus Bergvalls Foundation/ ; }, mesh = {Humans ; Molecular Chaperones/*metabolism ; Neurodegenerative Diseases/*genetics/pathology ; }, abstract = {The dysfunction of the proteostasis network is a molecular hallmark of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. Molecular chaperones are a major component of the proteostasis network and maintain cellular homeostasis by folding client proteins, assisting with intracellular transport, and interfering with protein aggregation or degradation. Heat shock protein 70 kDa (Hsp70) and 90 kDa (Hsp90) are two of the most important chaperones whose functions are dependent on ATP hydrolysis and collaboration with their co-chaperones. Numerous studies implicate Hsp70, Hsp90, and their co-chaperones in neurodegenerative diseases. Targeting the specific protein-protein interactions between chaperones and their particular partner co-chaperones with small molecules provides an opportunity to specifically modulate Hsp70 or Hsp90 function for neurodegenerative diseases. Here, we review the roles of co-chaperones in Hsp70 or Hsp90 chaperone cycles, the impacts of co-chaperones in neurodegenerative diseases, and the development of small molecules modulating chaperone/co-chaperone interactions. We also provide a future perspective of drug development targeting chaperone/co-chaperone interactions for neurodegenerative diseases.}, }
@article {pmid34739333, year = {2021}, author = {Gwon, Y and Maxwell, BA and Kolaitis, RM and Zhang, P and Kim, HJ and Taylor, JP}, title = {Ubiquitination of G3BP1 mediates stress granule disassembly in a context-specific manner.}, journal = {Science (New York, N.Y.)}, volume = {372}, number = {6549}, pages = {eabf6548}, pmid = {34739333}, issn = {1095-9203}, support = {R35 NS097974/NS/NINDS NIH HHS/United States ; R35NS097074/NH/NIH HHS/United States ; 18-IIA-419//Amyotrophic Lateral Sclerosis Association/ ; /HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Autophagy ; Blood Proteins/*metabolism ; Cell Line, Tumor ; Cytoplasmic Granules/*metabolism ; DNA Helicases/chemistry/genetics/*metabolism ; Endoplasmic Reticulum/metabolism ; HEK293 Cells ; *Heat-Shock Response ; Humans ; Intracellular Membranes/metabolism ; Membrane Proteins/*metabolism ; Mutation ; Poly-ADP-Ribose Binding Proteins/chemistry/genetics/*metabolism ; Polyubiquitin/metabolism ; Protein Domains ; Proteolysis ; RNA Helicases/chemistry/genetics/*metabolism ; RNA Recognition Motif Proteins/chemistry/genetics/*metabolism ; Ubiquitinated Proteins/chemistry/*metabolism ; Ubiquitination ; Valosin Containing Protein/*metabolism ; }, abstract = {Stress granules are dynamic, reversible condensates composed of RNA and protein that assemble in eukaryotic cells in response to a variety of stressors and are normally disassembled after stress is removed. The composition and assembly of stress granules is well understood, but little is known about the mechanisms that govern disassembly. Impaired disassembly has been implicated in some diseases including amyotrophic lateral sclerosis, frontotemporal dementia, and multisystem proteinopathy. Using cultured human cells, we found that stress granule disassembly was context-dependent: Specifically in the setting of heat shock, disassembly required ubiquitination of G3BP1, the central protein within the stress granule RNA-protein network. We found that ubiquitinated G3BP1 interacted with the endoplasmic reticulum–associated protein FAF2, which engaged the ubiquitin-dependent segregase p97/VCP (valosin-containing protein). Thus, targeting of G3BP1 weakened the stress granule–specific interaction network, resulting in granule disassembly.}, }
@article {pmid35101542, year = {2022}, author = {Keating, SS and San Gil, R and Swanson, MEV and Scotter, EL and Walker, AK}, title = {TDP-43 pathology: From noxious assembly to therapeutic removal.}, journal = {Progress in neurobiology}, volume = {211}, number = {}, pages = {102229}, doi = {10.1016/j.pneurobio.2022.102229}, pmid = {35101542}, issn = {1873-5118}, mesh = {Amyotrophic Lateral Sclerosis/metabolism ; *DNA-Binding Proteins/chemistry/metabolism ; Frontotemporal Dementia ; Humans ; Protein Folding ; *TDP-43 Proteinopathies/metabolism ; }, abstract = {Our understanding of amyotrophic lateral sclerosis and frontotemporal dementia has advanced dramatically since the discovery of cytoplasmic TAR DNA-binding protein 43 (TDP-43) inclusions as the hallmark pathology of these neurodegenerative diseases. Recent studies have provided insights into the physiological function of TDP-43 as an essential DNA-/RNA-modulating protein, and the triggers and consequences of TDP-43 dysfunction and aggregation. The formation of TDP-43 pathology is a progressive process, involving the generation of multiple distinct protein species, each with varying biophysical properties and roles in neurodegeneration. Here, we explore how the pathogenic changes to TDP-43, including mislocalisation, misfolding, aberrant liquid-liquid phase separation, stress granule assembly, oligomerisation, and post-translational modification, drive disease-associated aggregation in TDP-43 proteinopathies. We highlight how pathological TDP-43 species are formed and contribute to cellular dysfunction and toxicity, via both loss-of-function and gain-of-function mechanisms. We also review the role of protein homeostasis mechanisms, namely the ubiquitin proteasome system, autophagy-lysosome pathway, heat-shock response, and chaperone-mediated autophagy, in combating TDP-43 aggregation and discuss how their dysfunction likely promotes disease pathogenesis and progression. Finally, we evaluate pre-clinical studies aimed at enhancing TDP-43 protein clearance via these mechanisms and provide insight on promising strategies for future therapeutic advances. Harnessing the mechanisms that protect against or ameliorate TDP-43 pathology presents promising opportunities for developing disease-modifying treatments for these neurodegenerative diseases.}, }
@article {pmid35159325, year = {2022}, author = {Casarotto, E and Sproviero, D and Corridori, E and Gagliani, MC and Cozzi, M and Chierichetti, M and Cristofani, R and Ferrari, V and Galbiati, M and Mina, F and Piccolella, M and Rusmini, P and Tedesco, B and Gagliardi, S and Cortese, K and Cereda, C and Poletti, A and Crippa, V}, title = {Neurodegenerative Disease-Associated TDP-43 Fragments Are Extracellularly Secreted with CASA Complex Proteins.}, journal = {Cells}, volume = {11}, number = {3}, pages = {}, pmid = {35159325}, issn = {2073-4409}, support = {2017_0747//Fondazione Cariplo/ ; 2017F2A2C5//Ministry of Education, Universities and Research/ ; PSR2020//University of Milan/ ; SEED 2019: #TDP-43-iPSC//University of Milan/ ; AFM Telethon n. 16406//Association Fran&#xe7;aise contre les Myopathies/ ; Grant Heavy Equipment D.R. 3404//University of Genoa/ ; }, mesh = {Adaptor Proteins, Signal Transducing/metabolism ; *Amyotrophic Lateral Sclerosis/metabolism ; Apoptosis Regulatory Proteins/metabolism ; Autophagy-Related Proteins/metabolism ; *DNA-Binding Proteins/metabolism ; *Extracellular Vesicles/metabolism ; *Frontotemporal Lobar Degeneration ; Humans ; Molecular Chaperones/metabolism ; *Neurodegenerative Diseases ; Peptide Fragments/metabolism ; }, abstract = {Extracellular vesicles (EVs) play a central role in neurodegenerative diseases (NDs) since they may either spread the pathology or contribute to the intracellular protein quality control (PQC) system for the cellular clearance of NDs-associated proteins. Here, we investigated the crosstalk between large (LVs) and small (SVs) EVs and PQC in the disposal of TDP-43 and its FTLD and ALS-associated C-terminal fragments (TDP-35 and TDP-25). By taking advantage of neuronal cells (NSC-34 cells), we demonstrated that both EVs types, but particularly LVs, contained TDP-43, TDP-35 and TDP-25. When the PQC system was inhibited, as it occurs in NDs, we found that TDP-35 and TDP-25 secretion via EVs increased. In line with this observation, we specifically detected TDP-35 in EVs derived from plasma of FTLD patients. Moreover, we demonstrated that both neuronal and plasma-derived EVs transported components of the chaperone-assisted selective autophagy (CASA) complex (HSP70, BAG3 and HSPB8). Neuronal EVs also contained the autophagy-related MAP1LC3B-II protein. Notably, we found that, under PQC inhibition, HSPB8, BAG3 and MAP1LC3B-II secretion paralleled that of TDP-43 species. Taken together, our data highlight the role of EVs, particularly of LVs, in the disposal of disease-associated TDP-43 species, and suggest a possible new role for the CASA complex in NDs.}, }
@article {pmid35163711, year = {2022}, author = {Clarke, BE and Kalmar, B and Greensmith, L}, title = {Enhanced Expression of TRAP1 Protects Mitochondrial Function in Motor Neurons under Conditions of Oxidative Stress.}, journal = {International journal of molecular sciences}, volume = {23}, number = {3}, pages = {}, pmid = {35163711}, issn = {1422-0067}, mesh = {Animals ; Mice ; *Amyotrophic Lateral Sclerosis/genetics/metabolism ; HSP90 Heat-Shock Proteins/genetics/metabolism ; Mitochondria/genetics/metabolism ; Motor Neurons/metabolism ; *Neurodegenerative Diseases/metabolism ; Oxidative Stress ; Primary Cell Culture ; }, abstract = {TNF-receptor associated protein (TRAP1) is a cytoprotective mitochondrial-specific member of the Hsp90 heat shock protein family of protein chaperones that has been shown to antagonise mitochondrial apoptosis and oxidative stress, regulate the mitochondrial permeability transition pore and control protein folding in mitochondria. Here we show that overexpression of TRAP1 protects motor neurons from mitochondrial dysfunction and death induced by exposure to oxidative stress conditions modelling amyotrophic lateral sclerosis (ALS). ALS is a fatal neurodegenerative disease in which motor neurons degenerate, leading to muscle weakness and atrophy and death, typically within 3 years of diagnosis. In primary murine motor neurons, shRNA-mediated knockdown of TRAP1 expression results in mitochondrial dysfunction but does not further exacerbate damage induced by oxidative stress alone. Together, these results show that TRAP1 may be a potential therapeutic target for neurodegenerative diseases such as ALS, where mitochondrial dysfunction has been shown to be an early marker of pathogenesis.}, }
@article {pmid35164882, year = {2022}, author = {Liu, F and Morderer, D and Wren, MC and Vettleson-Trutza, SA and Wang, Y and Rabichow, BE and Salemi, MR and Phinney, BS and Oskarsson, B and Dickson, DW and Rossoll, W}, title = {Proximity proteomics of C9orf72 dipeptide repeat proteins identifies molecular chaperones as modifiers of poly-GA aggregation.}, journal = {Acta neuropathologica communications}, volume = {10}, number = {1}, pages = {22}, pmid = {35164882}, issn = {2051-5960}, support = {/HHMI/Howard Hughes Medical Institute/United States ; P30 AG062677/AG/NIA NIH HHS/United States ; R33 NS110960/NS/NINDS NIH HHS/United States ; NS084974/AG/NIA NIH HHS/United States ; P01 NS084974/NS/NINDS NIH HHS/United States ; }, mesh = {*Amyotrophic Lateral Sclerosis ; *C9orf72 Protein ; Dipeptides ; *Frontotemporal Dementia ; HEK293 Cells ; Humans ; Molecular Chaperones ; *Protein Aggregation, Pathological ; Proteomics ; RNA ; *Repetitive Sequences, Nucleic Acid ; }, abstract = {The most common inherited cause of two genetically and clinico-pathologically overlapping neurodegenerative diseases, amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), is the presence of expanded GGGGCC intronic hexanucleotide repeats in the C9orf72 gene. Aside from haploinsufficiency and toxic RNA foci, another non-exclusive disease mechanism is the non-canonical translation of the repeat RNA into five different dipeptide repeat proteins (DPRs), which form neuronal inclusions in affected patient brains. While evidence from cellular and animal models supports a toxic gain-of-function of pathologic poly-GA, poly-GR, and poly-PR aggregates in promoting deposition of TDP-43 pathology and neurodegeneration in affected brain areas, the relative contribution of DPRs to the disease process in c9FTD/ALS patients remains unclear. Here we have used the proximity-dependent biotin identification (BioID) proximity proteomics approach to investigate the formation and collective composition of DPR aggregates using cellular models. While interactomes of arginine rich poly-GR and poly-PR aggregates overlapped and were enriched for nucleolar and ribosomal proteins, poly-GA aggregates demonstrated a distinct association with proteasomal components, molecular chaperones (HSPA1A/HSP70, HSPA8/HSC70, VCP/p97), co-chaperones (BAG3, DNAJA1A) and other factors that regulate protein folding and degradation (SQSTM1/p62, CALR, CHIP/STUB1). Experiments in cellular models of poly-GA pathology show that molecular chaperones and co-chaperones are sequestered to the periphery of dense cytoplasmic aggregates, causing depletion from their typical cellular localization. Their involvement in the pathologic process is confirmed in autopsy brain tissue, where HSPA8, BAG3, VCP, and its adapter protein UBXN6 show a close association with poly-GA aggregates in the frontal cortex, temporal cortex, and hippocampus of c9FTLD and c9ALS cases. The association of heat shock proteins and co-chaperones with poly-GA led us to investigate their potential role in reducing its aggregation. We identified HSP40 co-chaperones of the DNAJB family as potent modifiers that increased the solubility of poly-GA, highlighting a possible novel therapeutic avenue and a central role of molecular chaperones in the pathogenesis of human C9orf72-linked diseases.}, }
@article {pmid35170035, year = {2022}, author = {Claesson, K and Chew, YL and Ecroyd, H}, title = {Exploiting flow cytometry for the unbiased quantification of protein inclusions in Caenorhabditis elegans.}, journal = {Journal of neurochemistry}, volume = {161}, number = {3}, pages = {281-292}, pmid = {35170035}, issn = {1471-4159}, support = {P40 OD010440/OD/NIH HHS/United States ; }, mesh = {Aging ; Animals ; *Caenorhabditis elegans ; Flow Cytometry ; *Inclusion Bodies ; Microscopy, Fluorescence ; }, abstract = {The aggregation of proteins into inclusions or plaques is a prominent hallmark of a diverse range of pathologies including neurodegenerative diseases. The quantification of such inclusions in Caenorhabditis elegans models of aggregation is usually achieved by fluorescence microscopy or other techniques involving biochemical fractionation of worm lysates. Here, we describe a simple and rapid flow cytometry-based approach that allows fluorescently tagged inclusions to be enumerated in whole worm lysate in a quantitative and unbiased fashion. We demonstrate that this technique is applicable to multiple C. elegans models of aggregation and importantly, can be used to monitor the dynamics of inclusion formation in response to heat shock and during ageing. This includes the characterisation of physicochemical properties of inclusions, such as their apparent size, which may reveal how aggregate formation is distinct in different tissues or at different stages of pathology or ageing. This new method can be used as a powerful technique for the medium- to high-throughput quantification of inclusions in future studies of genetic or chemical modulators of aggregation in C. elegans.}, }
@article {pmid35185565, year = {2022}, author = {Ueda, T and Ito, T and Inden, M and Kurita, H and Yamamoto, A and Hozumi, I}, title = {Stem Cells From Human Exfoliated Deciduous Teeth-Conditioned Medium (SHED-CM) is a Promising Treatment for Amyotrophic Lateral Sclerosis.}, journal = {Frontiers in pharmacology}, volume = {13}, number = {}, pages = {805379}, pmid = {35185565}, issn = {1663-9812}, abstract = {Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder, characterized by the loss of upper and lower motor neurons, for which an effective treatment has yet to be developed. Previous reports have shown that excessive oxidative stress, related to mitochondrial dysfunction and the accumulation of misfolding protein, contributes to ALS pathology. In terms of treatment, it remains necessary to identify effective medicines for multiple therapeutic targets and have additive effects against several disorders. In this study, we investigated stem cells from human exfoliated deciduous teeth (SHED), which release many factors, such as neurotrophic factors and cytokines, and are applied to treat neurological diseases. Specifically, we examined whether SHED-conditioned medium (CM), i.e., the serum-free culture supernatant of SHED, reduced mutant SOD1-induced intracellular aggregates and neurotoxicity. We found that SHED-CM significantly suppressed the mutant SOD1-induced intracellular aggregates and neurotoxicity. The neuroprotective effects of SHED-CM are partly related to heat shock protein and the activation of insulin-like growth factor-1 receptor. SHED-CM also had a protective effect on induced pluripotent stem cell-derived motor neurons. Moreover, SHED-CM was effective against not only familial ALS but also sporadic ALS. Overall, these results suggest that SHED-CM could be a promising treatment for slowing the progression of ALS.}, }
@article {pmid35247097, year = {2022}, author = {Mohan, HM and Trzeciakiewicz, H and Pithadia, A and Crowley, EV and Pacitto, R and Safren, N and Trotter, B and Zhang, C and Zhou, X and Zhang, Y and Basrur, V and Paulson, HL and Sharkey, LM}, title = {RTL8 promotes nuclear localization of UBQLN2 to subnuclear compartments associated with protein quality control.}, journal = {Cellular and molecular life sciences : CMLS}, volume = {79}, number = {3}, pages = {176}, pmid = {35247097}, issn = {1420-9071}, support = {P30 AG053760/AG/NIA NIH HHS/United States ; 1P30AG053760-01/NH/NIH HHS/United States ; R21 NS093469/NS/NINDS NIH HHS/United States ; R35 NS122302/NS/NINDS NIH HHS/United States ; 9R01NS096785-06/NH/NIH HHS/United States ; R01 NS096785/NS/NINDS NIH HHS/United States ; }, mesh = {Adaptor Proteins, Signal Transducing/deficiency/genetics/*metabolism ; Amino Acid Sequence ; Animals ; Autophagy-Related Proteins/deficiency/genetics/metabolism ; Brain/metabolism ; Carrier Proteins/genetics/metabolism ; Cell Nucleolus/metabolism ; HEK293 Cells ; Humans ; Membrane Proteins/chemistry/genetics/*metabolism ; Mice ; Mice, Knockout ; Nuclear Proteins/genetics/metabolism ; Protein Binding ; Protein Isoforms/chemistry/genetics/metabolism ; Recombinant Proteins/biosynthesis/chemistry/isolation &amp; purification ; Sequence Alignment ; Temperature ; Ubiquitin/metabolism ; }, abstract = {The brain-expressed ubiquilins (UBQLNs) 1, 2 and 4 are a family of ubiquitin adaptor proteins that participate broadly in protein quality control (PQC) pathways, including the ubiquitin proteasome system (UPS). One family member, UBQLN2, has been implicated in numerous neurodegenerative diseases including ALS/FTD. UBQLN2 typically resides in the cytoplasm but in disease can translocate to the nucleus, as in Huntington's disease where it promotes the clearance of mutant Huntingtin. How UBQLN2 translocates to the nucleus and clears aberrant nuclear proteins, however, is not well understood. In a mass spectrometry screen to discover UBQLN2 interactors, we identified a family of small (13 kDa), highly homologous uncharacterized proteins, RTL8, and confirmed the interaction between UBQLN2 and RTL8 both in vitro using recombinant proteins and in vivo using mouse brain tissue. Under endogenous and overexpressed conditions, RTL8 localizes to nucleoli. When co-expressed with UBQLN2, RTL8 promotes nuclear translocation of UBQLN2. RTL8 also facilitates UBQLN2's nuclear translocation during heat shock. UBQLN2 and RTL8 colocalize within ubiquitin-enriched subnuclear structures containing PQC components. The robust effect of RTL8 on the nuclear translocation and subnuclear localization of UBQLN2 does not extend to the other brain-expressed ubiquilins, UBQLN1 and UBQLN4. Moreover, compared to UBQLN1 and UBQLN4, UBQLN2 preferentially stabilizes RTL8 levels in human cell lines and in mouse brain, supporting functional heterogeneity among UBQLNs. As a novel UBQLN2 interactor that recruits UBQLN2 to specific nuclear compartments, RTL8 may regulate UBQLN2 function in nuclear protein quality control.}, }
@article {pmid35507432, year = {2023}, author = {Wang, SM and Wu, HE and Yasui, Y and Geva, M and Hayden, M and Maurice, T and Cozzolino, M and Su, TP}, title = {Nucleoporin POM121 signals TFEB-mediated autophagy via activation of SIGMAR1/sigma-1 receptor chaperone by pridopidine.}, journal = {Autophagy}, volume = {19}, number = {1}, pages = {126-151}, pmid = {35507432}, issn = {1554-8635}, support = {K02 DA000206/DA/NIDA NIH HHS/United States ; Z01 DA000206/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Mice ; *Amyotrophic Lateral Sclerosis/metabolism ; *Autophagy/genetics ; C9orf72 Protein/genetics ; *Frontotemporal Dementia/genetics ; Heat-Shock Proteins/metabolism ; *Membrane Proteins/genetics ; Nuclear Pore Complex Proteins ; *Receptors, sigma/metabolism ; Sigma-1 Receptor ; Transcription Factors/metabolism ; Cell Line ; Piperidines ; }, abstract = {Macroautophagy/autophagy is an essential process for cellular survival and is implicated in many diseases. A critical step in autophagy is the transport of the transcription factor TFEB from the cytosol into the nucleus, through the nuclear pore (NP) by KPNB1/importinβ1. In the C9orf72 subtype of amyotrophic lateral sclerosis-frontotemporal lobar degeneration (ALS-FTD), the hexanucleotide (G4C2)RNA expansion (HRE) disrupts the nucleocytoplasmic transport of TFEB, compromising autophagy. Here we show that a molecular chaperone, the SIGMAR1/Sigma-1 receptor (sigma non-opioid intracellular receptor 1), facilitates TFEB transport into the nucleus by chaperoning the NP protein (i.e., nucleoporin) POM121 which recruits KPNB1. In NSC34 cells, HRE reduces TFEB transport by interfering with the association between SIGMAR1 and POM121, resulting in reduced nuclear levels of TFEB, KPNB1, and the autophagy marker LC3-II. Overexpression of SIGMAR1 or POM121, or treatment with the highly selective and potent SIGMAR1 agonist pridopidine, currently in phase 2/3 clinical trials for ALS and Huntington disease, rescues all of these deficits. Our results implicate nucleoporin POM121 not merely as a structural nucleoporin, but also as a chaperone-operated signaling molecule enabling TFEB-mediated autophagy. Our data suggest the use of SIGMAR1 agonists, such as pridopidine, for therapeutic development of diseases in which autophagy is impaired.Abbreviations: ALS-FTD, amyotrophic lateral sclerosis-frontotemporal dementiaC9ALS-FTD, C9orf72 subtype of amyotrophic lateral sclerosis-frontotemporal dementiaCS, citrate synthaseER, endoplasmic reticulumGSS, glutathione synthetaseHRE, hexanucleotide repeat expansionHSPA5/BiP, heat shock protein 5LAMP1, lysosomal-associated membrane protein 1MAM, mitochondria-associated endoplasmic reticulum membraneMAP1LC3/LC3, microtubule-associated protein 1 light chain 3NP, nuclear poreNSC34, mouse motor neuron-like hybrid cell lineNUPs, nucleoporinsPOM121, nuclear pore membrane protein 121SIGMAR1/Sigma-1R, sigma non-opioid intracellular receptor 1TFEB, transcription factor EBTMEM97/Sigma-2R, transmembrane protein 97.}, }
@article {pmid35521960, year = {2023}, author = {Cheng, X and Zhang, P and Zhao, H and Zheng, H and Zheng, K and Zhang, H and Zhang, H}, title = {Proteotoxic stress disrupts epithelial integrity by inducing MTOR sequestration and autophagy overactivation.}, journal = {Autophagy}, volume = {19}, number = {1}, pages = {241-255}, pmid = {35521960}, issn = {1554-8635}, support = {P40 OD010440/OD/NIH HHS/United States ; }, mesh = {Animals ; *Autophagy ; Caenorhabditis elegans/metabolism ; Proteotoxic Stress ; Actins/metabolism ; Drosophila/metabolism ; TOR Serine-Threonine Kinases/metabolism ; Lysosomes/metabolism ; *Caenorhabditis elegans Proteins/metabolism ; Basic Helix-Loop-Helix Proteins/metabolism ; }, abstract = {Macroautophagy/autophagy, an evolutionarily conserved degradation system, serves to clear intracellular components through the lysosomal pathway. Mounting evidence has revealed cytoprotective roles of autophagy; however, the intracellular causes of overactivated autophagy, which has cytotoxic effects, remain elusive. Here we show that sustained proteotoxic stress induced by loss of the RING and Kelch repeat-containing protein C53A5.6/RIKE-1 induces sequestration of LET-363/MTOR complex and overactivation of autophagy, and consequently impairs epithelial integrity in C. elegans. In C53A5.6/RIKE-1-deficient animals, blocking autophagosome formation effectively prevents excessive endosomal degradation, mitigates mislocalization of intestinal membrane components and restores intestinal lumen morphology. However, autophagy inhibition does not affect LET-363/MTOR aggregation in animals with compromised C53A5.6/RIKE-1 function. Improving proteostasis capacity by reducing DAF-2 insulin/IGF1 signaling markedly relieves the aggregation of LET-363/MTOR and alleviates autophagy overactivation, which in turn reverses derailed endosomal trafficking and rescues epithelial morphogenesis defects in C53A5.6/RIKE-1-deficient animals. Hence, our studies reveal that C53A5.6/RIKE-1-mediated proteostasis is critical for maintaining the basal level of autophagy and epithelial integrity.Abbreviations: ACT-5: actin 5; ACTB: actin beta; ALs: autolysosomes; APs: autophagosomes; AJM-1: apical junction molecule; ATG: autophagy related; C. elegans: Caenorhabditis elegans; CPL-1: cathepsin L family; DAF: abnormal dauer formation; DLG-1: Drosophila discs large homolog; ERM-1: ezrin/radixin/moesin; EPG: ectopic P granule; GFP: freen fluorescent protein; HLH-30: helix loop helix; HSP: heat shock protein; LAAT-1: lysosome associated amino acid transporter; LET: lethal; LGG-1: LC3, GABARAP and GATE-16 family; LMP-1: LAMP (lysosome-associated membrane protein) homolog; MTOR: mechanistic target of rapamycin kinase; NUC-1: abnormal nuclease; PEPT-1/OPT-2: Peptide transporter family; PGP-1: P-glycoprotein related; RAB: RAB family; RIKE-1: RING and Kelch repeat-containing protein; SLCF-1: solute carrier family; SQST-1: sequestosome related; SPTL-1: serine palmitoyl transferase family.}, }
@article {pmid35581326, year = {2022}, author = {François-Moutal, L and Scott, DD and Ambrose, AJ and Zerio, CJ and Rodriguez-Sanchez, M and Dissanayake, K and May, DG and Carlson, JM and Barbieri, E and Moutal, A and Roux, KJ and Shorter, J and Khanna, R and Barmada, SJ and McGurk, L and Khanna, M}, title = {Heat shock protein Grp78/BiP/HspA5 binds directly to TDP-43 and mitigates toxicity associated with disease pathology.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {8140}, pmid = {35581326}, issn = {2045-2322}, support = {P30 AG053760/AG/NIA NIH HHS/United States ; R01 NS097542/NS/NINDS NIH HHS/United States ; R35GM126949/NH/NIH HHS/United States ; P20 GM103620/GM/NIGMS NIH HHS/United States ; R35 GM126949/GM/NIGMS NIH HHS/United States ; P30 AG072931/AG/NIA NIH HHS/United States ; R21 AG065854/AG/NIA NIH HHS/United States ; R01NS097542/NH/NIH HHS/United States ; R01 NS113943/NS/NINDS NIH HHS/United States ; }, mesh = {*Amyotrophic Lateral Sclerosis/metabolism ; Animals ; DNA-Binding Proteins/metabolism ; Drosophila/metabolism ; Endoplasmic Reticulum Chaperone BiP ; HSP70 Heat-Shock Proteins/genetics ; Heat-Shock Proteins/metabolism ; Humans ; Molecular Chaperones ; *Neurodegenerative Diseases ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with no cure or effective treatment in which TAR DNA Binding Protein of 43 kDa (TDP-43) abnormally accumulates into misfolded protein aggregates in affected neurons. It is widely accepted that protein misfolding and aggregation promotes proteotoxic stress. The molecular chaperones are a primary line of defense against proteotoxic stress, and there has been long-standing interest in understanding the relationship between chaperones and aggregated protein in ALS. Of particular interest are the heat shock protein of 70 kDa (Hsp70) family of chaperones. However, defining which of the 13 human Hsp70 isoforms is critical for ALS has presented many challenges. To gain insight into the specific Hsp70 that modulates TDP-43, we investigated the relationship between TDP-43 and the Hsp70s using proximity-dependent biotin identification (BioID) and discovered several Hsp70 isoforms associated with TDP-43 in the nucleus, raising the possibility of an interaction with native TDP-43. We further found that HspA5 bound specifically to the RNA-binding domain of TDP-43 using recombinantly expressed proteins. Moreover, in a Drosophila strain that mimics ALS upon TDP-43 expression, the mRNA levels of the HspA5 homologue (Hsc70.3) were significantly increased. Similarly we observed upregulation of HspA5 in prefrontal cortex neurons from human ALS patients. Finally, overexpression of HspA5 in Drosophila rescued TDP-43-induced toxicity, suggesting that upregulation of HspA5 may have a compensatory role in ALS pathobiology.}, }
@article {pmid35701899, year = {2022}, author = {Watabe, K and Niida-Kawaguchi, M and Tada, M and Kato, Y and Murata, M and Tanji, K and Wakabayashi, K and Yamada, M and Kakita, A and Shibata, N}, title = {Praja1 RING-finger E3 ubiquitin ligase is a common suppressor of neurodegenerative disease-associated protein aggregation.}, journal = {Neuropathology : official journal of the Japanese Society of Neuropathology}, volume = {42}, number = {6}, pages = {488-504}, pmid = {35701899}, issn = {1440-1789}, support = {#18K06507//Japan Society for the Promotion of Science/ ; 2020-2021//Niigata University/ ; }, mesh = {*Amyotrophic Lateral Sclerosis/pathology ; *Frontotemporal Lobar Degeneration/pathology ; Heat Shock Transcription Factors ; *Neurodegenerative Diseases ; Protein Aggregates ; *Ubiquitin-Protein Ligases/genetics/metabolism ; Animals ; }, abstract = {The formation of misfolded protein aggregates is one of the pathological hallmarks of neurodegenerative diseases. We have previously demonstrated the cytoplasmic aggregate formation of adenovirally expressed transactivation response DNA-binding protein of 43 kDa (TDP-43), the main constituent of neuronal cytoplasmic aggregates in cases of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD), in cultured neuronal cells under the condition of proteasome inhibition. The TDP-43 aggregate formation was markedly suppressed by co-infection of adenoviruses expressing heat shock transcription factor 1 (HSF1), a master regulator of heat shock response, and Praja1 RING-finger E3 ubiquitin ligase (PJA1) located downstream of the HSF1 pathway. In the present study, we examined other reportedly known E3 ubiquitin ligases for TDP-43, i.e. Parkin, RNF112 and RNF220, but failed to find their suppressive effects on neuronal cytoplasmic TDP-43 aggregate formation, although they all bind to TDP-43 as verified by co-immunoprecipitation. In contrast, PJA1 also binds to adenovirally expressed wild-type and mutated fused in sarcoma, superoxide dismutase 1, α-synuclein and ataxin-3, and huntingtin polyglutamine proteins in neuronal cultures and suppressed the aggregate formation of these proteins. These results suggest that PJA1 is a common sensing factor for aggregate-prone proteins to counteract their aggregation propensity, and could be a potential therapeutic target for neurodegenerative diseases that include ALS, FTLD, Parkinson's disease and polyglutamine diseases.}, }
@article {pmid35713146, year = {2022}, author = {Azari, A and Goodarzi, A and Jafarkhani, B and Eghbali, M and Karimi, Z and Hosseini Balef, SS and Irannejad, H}, title = {Novel Molecular Targets and Mechanisms for Neuroprotective Modulation in Neurodegenerative Disorders.}, journal = {Central nervous system agents in medicinal chemistry}, volume = {22}, number = {2}, pages = {88-107}, doi = {10.2174/1871524922666220616092132}, pmid = {35713146}, issn = {1875-6166}, mesh = {Autophagy/genetics ; Humans ; *Neurodegenerative Diseases/drug therapy/metabolism ; Neurons/metabolism ; *alpha-Synuclein/metabolism ; }, abstract = {BACKGROUND: Neuronal death underlies the symptoms of several human neurological disorders, including Alzheimer's, Parkinson's and Huntington's diseases, and amyotrophic lateral sclerosis and their precise pathophysiology have not yet been elucidated. According to various studies, the prohibition is the best therapy with neuroprotective approaches, which are advanced and safe methods.<br><br>METHODS: This review summarizes some of the already-known and newly emerged neuroprotective targets and strategies and their experimental effects have also been reported. Accordingly, literature was studied from 2000 to 2021, and appropriate articles were searched in Google Scholar and Scopus with the keywords given in the keywords section of the current review.<br><br>RESULTS: Lewy bodies are the histopathologic characteristics of neurodegenerative disorders and are protein-rich intracellular deposits in which Alpha-synuclein is its major protein. Alphasynuclein's toxic potential provides a compelling rationale for therapeutic strategies aimed at decreasing its burden in neuronal cells through numerous pathways, including ubiquitin-proteasome system and autophagy-lysosome pathway, proteolytic breakdown via cathepsin D, kallikrein-6 (neurosin), calpain-1 or MMP9, heat shock proteins, and proteolysis targeting chimera which consists of a target protein-ligand and an E3 ubiquitin ligase (E3) followed by target protein ubiquitination (PROTACs). Other targets that have been noticed recently are the mutant huntingtin, tau proteins and glycogen synthase kinase 3&#x3b2;; their accumulation proceeds extensive neuronal damage and up to the minute approach such as proteolysis targeting chimera promotes its degradation in cells. Various studies demonstrated that Mendelian gene mutations can result in neurodegenerative diseases. An additional target that has gained much interest is epigenetics, such as mutation, phosphodiesterase, RNA binding proteins and Nuclear respiratory factor 1.<br><br>CONCLUSION: The novel molecular targets and new strategies compiled and introduced here can be used by scientists to design and discover more efficient small molecule drugs against neurodegenerative diseases. And also, the genes in which their mutations can lead to the &#x3b1;-synuclein aggregation or accumulation have been discussed and considered a valuable information on epigenetics in dementia.}, }
@article {pmid35787375, year = {2022}, author = {Scoles, DR and Gandelman, M and Paul, S and Dexheimer, T and Dansithong, W and Figueroa, KP and Pflieger, LT and Redlin, S and Kales, SC and Sun, H and Maloney, D and Damoiseaux, R and Henderson, MJ and Simeonov, A and Jadhav, A and Pulst, SM}, title = {A quantitative high-throughput screen identifies compounds that lower expression of the SCA2-and ALS-associated gene ATXN2.}, journal = {The Journal of biological chemistry}, volume = {298}, number = {8}, pages = {102228}, pmid = {35787375}, issn = {1083-351X}, support = {R35 NS127253/NS/NINDS NIH HHS/United States ; P30 CA016042/CA/NCI NIH HHS/United States ; R56 NS033123/NS/NINDS NIH HHS/United States ; RC4 NS073009/NS/NINDS NIH HHS/United States ; R01 NS097903/NS/NINDS NIH HHS/United States ; R21 NS103009/NS/NINDS NIH HHS/United States ; R37 NS033123/NS/NINDS NIH HHS/United States ; U01 NS103883/NS/NINDS NIH HHS/United States ; }, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/drug therapy/genetics ; *Ataxin-2/genetics ; Cerebellum/metabolism ; Cytoskeletal Proteins/metabolism ; HEK293 Cells ; RNA-Binding Proteins/metabolism ; Sodium-Potassium-Exchanging ATPase/metabolism ; *Spinocerebellar Ataxias/drug therapy/genetics ; }, abstract = {CAG repeat expansions in the ATXN2 (ataxin-2) gene can cause the autosomal dominant disorder spinocerebellar ataxia type 2 (SCA2) as well as increase the risk of ALS. Abnormal molecular, motor, and neurophysiological phenotypes in SCA2 mouse models are normalized by lowering ATXN2 transcription, and reduction of nonmutant Atxn2 expression has been shown to increase the life span of mice overexpressing the TDP-43 (transactive response DNA-binding protein 43 kDa) ALS protein, demonstrating the potential benefits of targeting ATXN2 transcription in humans. Here, we describe a quantitative high-throughput screen to identify compounds that lower ATXN2 transcription. We screened 428,759 compounds in a multiplexed assay using an ATXN2-luciferase reporter in human embryonic kidney 293 (HEK-293) cells and identified a diverse set of compounds capable of lowering ATXN2 transcription. We observed dose-dependent reductions of endogenous ATXN2 in HEK-293 cells treated with procillaridin A, 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG), and heat shock protein 990 (HSP990), known inhibitors of HSP90 and Na[+]/K[+]-ATPases. Furthermore, HEK-293 cells expressing polyglutamine-expanded ATXN2-Q58 treated with 17-DMAG had minimally detectable ATXN2, as well as normalized markers of autophagy and endoplasmic reticulum stress, including STAU1 (Staufen 1), molecular target of rapamycin, p62, LC3-II (microtubule-associated protein 1A/1B-light chain 3II), CHOP (C/EBP homologous protein), and phospho-eIF2α (eukaryotic initiation factor 2α). Finally, bacterial artificial chromosome ATXN2-Q22 mice treated with 17-DMAG or HSP990 exhibited highly reduced ATXN2 protein abundance in the cerebellum. Taken together, our study demonstrates inhibition of HSP90 or Na[+]/K[+]-ATPases as potentially effective therapeutic strategies for treating SCA2 and ALS.}, }
@article {pmid36075972, year = {2022}, author = {Lu, S and Hu, J and Arogundade, OA and Goginashvili, A and Vazquez-Sanchez, S and Diedrich, JK and Gu, J and Blum, J and Oung, S and Ye, Q and Yu, H and Ravits, J and Liu, C and Yates, JR and Cleveland, DW}, title = {Heat-shock chaperone HSPB1 regulates cytoplasmic TDP-43 phase separation and liquid-to-gel transition.}, journal = {Nature cell biology}, volume = {24}, number = {9}, pages = {1378-1393}, pmid = {36075972}, issn = {1476-4679}, support = {P41 GM103533/GM/NIGMS NIH HHS/United States ; P30 NS047101/NS/NINDS NIH HHS/United States ; R01 NS027036/NS/NINDS NIH HHS/United States ; R00 NS114162/NS/NINDS NIH HHS/United States ; R01 NS121604/NS/NINDS NIH HHS/United States ; R01 NS088578/NS/NINDS NIH HHS/United States ; }, mesh = {Adenosine Triphosphate ; Amyotrophic Lateral Sclerosis/genetics/metabolism ; *DNA-Binding Proteins/chemistry/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; *Heat-Shock Proteins/chemistry/metabolism ; *Heat-Shock Proteins, Small ; Humans ; Molecular Chaperones/genetics ; *Phase Transition ; Proteasome Endopeptidase Complex ; RNA/metabolism ; }, abstract = {While acetylated, RNA-binding-deficient TDP-43 reversibly phase separates within nuclei into complex droplets (anisosomes) comprised of TDP-43-containing liquid outer shells and liquid centres of HSP70-family chaperones, cytoplasmic aggregates of TDP-43 are hallmarks of multiple neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). Here we show that transient oxidative stress, proteasome inhibition or inhibition of the ATP-dependent chaperone activity of HSP70 provokes reversible cytoplasmic TDP-43 de-mixing and transition from liquid to gel/solid, independently of RNA binding or stress granules. Isotope labelling mass spectrometry was used to identify that phase-separated cytoplasmic TDP-43 is bound by the small heat-shock protein HSPB1. Binding is direct, mediated through TDP-43's RNA binding and low-complexity domains. HSPB1 partitions into TDP-43 droplets, inhibits TDP-43 assembly into fibrils, and is essential for disassembly of stress-induced TDP-43 droplets. A decrease in HSPB1 promotes cytoplasmic TDP-43 de-mixing and mislocalization. HSPB1 depletion was identified in spinal motor neurons of patients with ALS containing aggregated TDP-43. These findings identify HSPB1 to be a regulator of cytoplasmic TDP-43 phase separation and aggregation.}, }
@article {pmid36423739, year = {2023}, author = {Phung, TH and Tatman, M and Monteiro, MJ}, title = {UBQLN2 undergoes a reversible temperature-induced conformational switch that regulates binding with HSPA1B: ALS/FTD mutations cripple the switch but do not destroy HSPA1B binding.}, journal = {Biochimica et biophysica acta. General subjects}, volume = {1867}, number = {2}, pages = {130284}, pmid = {36423739}, issn = {1872-8006}, support = {R01 NS098243/NS/NINDS NIH HHS/United States ; R01 NS100008/NS/NINDS NIH HHS/United States ; RF1 NS098243/NS/NINDS NIH HHS/United States ; }, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/genetics/metabolism ; *Frontotemporal Dementia/genetics ; Temperature ; Tryptophan/genetics ; Adaptor Proteins, Signal Transducing/genetics/metabolism ; Autophagy-Related Proteins/genetics/metabolism ; Mutation ; HSP70 Heat-Shock Proteins/genetics ; }, abstract = {Here we present evidence, based on alterations of its intrinsic tryptophan fluorescence, that UBQLN2 protein undergoes a conformational switch when the temperature is raised from 37 °C to 42 °C. The switch is reset on restoration of the temperature. We speculate that the switch regulates UBQLN2 function in the heat shock response because elevation of the temperature from 37 °C to 42 °C dramatically increased in vitro binding between UBQLN2 and HSPA1B. Furthermore, restoration of the temperature to 37 °C decreased HSPA1B binding. By comparison to wild type (WT) UBQLN2, we found that all five ALS/FTD mutant UBQLN2 proteins we examined had attenuated alterations in tryptophan fluorescence when shifted to 42 °C, suggesting that the conformational switch is crippled in the mutants. Paradoxically, all five mutants bound similar amounts of HSPA1B compared to WT UBQLN2 protein at 42 °C, suggesting that either the conformational switch is not instrumental for HSPA1B binding, or that, although damaged, it is still functional. Comparison of the poly-ubiquitin chain binding revealed that WT UBQLN2 binds more avidly with K63 than with K48 chains. The avidity may explain the involvement of UBQLN2 in autophagy and cell signaling. Consistent with its function in autophagy, we found UBQLN2 binds directly with LC3, the autophagosomal-specific membrane-tethered protein. Finally, we provide evidence that WT UBQLN2 can homodimerize, and heterodimerize with WT UBQLN1. We show that ALS mutant P497S-UBQLN2 protein can oligomerize with either WT UBQLN1 or 2, providing a possible mechanism for how mutant UBQLN2 proteins could bind and inactivate UBQLN proteins, causing loss of function.}, }
@article {pmid36594740, year = {2023}, author = {Tedesco, B and Vendredy, L and Timmerman, V and Poletti, A}, title = {The chaperone-assisted selective autophagy complex dynamics and dysfunctions.}, journal = {Autophagy}, volume = {19}, number = {6}, pages = {1619-1641}, pmid = {36594740}, issn = {1554-8635}, mesh = {*Autophagy/genetics ; Kelch-Like ECH-Associated Protein 1/metabolism ; *NF-E2-Related Factor 2/metabolism ; Heat-Shock Proteins/metabolism ; Molecular Chaperones/metabolism ; Intracellular Signaling Peptides and Proteins/metabolism ; Ubiquitins/metabolism ; }, abstract = {Each protein must be synthesized with the correct amino acid sequence, folded into its native structure, and transported to a relevant subcellular location and protein complex. If any of these steps fail, the cell has the capacity to break down aberrant proteins to maintain protein homeostasis (also called proteostasis). All cells possess a set of well-characterized protein quality control systems to minimize protein misfolding and the damage it might cause. Autophagy, a conserved pathway for the degradation of long-lived proteins, aggregates, and damaged organelles, was initially characterized as a bulk degradation pathway. However, it is now clear that autophagy also contributes to intracellular homeostasis by selectively degrading cargo material. One of the pathways involved in the selective removal of damaged and misfolded proteins is chaperone-assisted selective autophagy (CASA). The CASA complex is composed of three main proteins (HSPA, HSPB8 and BAG3), essential to maintain protein homeostasis in muscle and neuronal cells. A failure in the CASA complex, caused by mutations in the respective coding genes, can lead to (cardio)myopathies and neurodegenerative diseases. Here, we summarize our current understanding of the CASA complex and its dynamics. We also briefly discuss how CASA complex proteins are involved in disease and may represent an interesting therapeutic target.Abbreviation ALP: autophagy lysosomal pathway; ALS: amyotrophic lateral sclerosis; AMOTL1: angiomotin like 1; ARP2/3: actin related protein 2/3; BAG: BAG cochaperone; BAG3: BAG cochaperone 3; CASA: chaperone-assisted selective autophagy; CMA: chaperone-mediated autophagy; DNAJ/HSP40: DnaJ heat shock protein family (Hsp40); DRiPs: defective ribosomal products; EIF2A/eIF2α: eukaryotic translation initiation factor 2A; EIF2AK1/HRI: eukaryotic translation initiation factor 2 alpha kinase 1; GABARAP: GABA type A receptor-associated protein; HDAC6: histone deacetylase 6; HSP: heat shock protein; HSPA/HSP70: heat shock protein family A (Hsp70); HSP90: heat shock protein 90; HSPB8: heat shock protein family B (small) member 8; IPV: isoleucine-proline-valine; ISR: integrated stress response; KEAP1: kelch like ECH associated protein 1; LAMP2A: lysosomal associated membrane protein 2A; LATS1: large tumor suppressor kinase 1; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MTOC: microtubule organizing center; MTOR: mechanistic target of rapamycin kinase; NFKB/NF-κB: nuclear factor kappa B; NFE2L2: NFE2 like bZIP transcription factor 2; PLCG/PLCγ: phospholipase C gamma; polyQ: polyglutamine; PQC: protein quality control; PxxP: proline-rich; RAN translation: repeat-associated non-AUG translation; SG: stress granule; SOD1: superoxide dismutase 1; SQSTM1/p62: sequestosome 1; STUB1/CHIP: STIP1 homology and U-box containing protein 1; STK: serine/threonine kinase; SYNPO: synaptopodin; TBP: TATA-box binding protein; TARDBP/TDP-43: TAR DNA binding protein; TFEB: transcription factor EB; TPR: tetratricopeptide repeats; TSC1: TSC complex subunit 1; UBA: ubiquitin associated; UPS: ubiquitin-proteasome system; WW: tryptophan-tryptophan; WWTR1: WW domain containing transcription regulator 1; YAP1: Yes1 associated transcriptional regulator.}, }
@article {pmid36687517, year = {2022}, author = {Yamashita, H and Komine, O and Fujimori-Tonou, N and Yamanaka, K}, title = {Comprehensive expression analysis with cell-type-specific transcriptome in ALS-linked mutant SOD1 mice: Revisiting the active role of glial cells in disease.}, journal = {Frontiers in cellular neuroscience}, volume = {16}, number = {}, pages = {1045647}, pmid = {36687517}, issn = {1662-5102}, abstract = {Non-cell autonomous mechanisms are involved in the pathogenesis of amyotrophic lateral sclerosis (ALS), an adult neurodegenerative disease characterized by selective motor neuron loss. While the emerging role of glial cells in ALS has been noted, the detailed cell-type-specific role of glial cells has not been clarified. Here, we examined mRNA expression changes using microarrays of the spinal cords of three distinct lines of mutant superoxide dismutase (SOD) 1 transgenic mice, an established ALS model. Our analysis used a transcriptome database of component cell types in the central nervous system (CNS), as well as SOD1 [G93A] cell-type transcriptomes. More than half of the differentially expressed genes (DEGs) were highly expressed in microglia, and enrichment analysis of DEGs revealed that immunological reactions were profoundly involved and some transcription factors were upregulated. Our analysis focused on DEGs that are highly expressed in each cell type, as well as chemokines, caspases, and heat shock proteins. Disease-associated microglial genes were upregulated, while homeostatic microglial genes were not, and galectin-3 (Mac2), a known activated microglial marker, was predicted to be ectopically expressed in astrocytes in mutant SOD1 mice. In mutant SOD1 mice, we developed a prediction model for the pathophysiology of different cell types related to TREM2, apolipoprotein E, and lipoproteins. Our analysis offers a viable resource to understand not only the molecular pathologies of each CNS constituent cell type, but also the cellular crosstalk between different cell types under both physiological and pathological conditions in model mice for various neurodegenerative diseases.}, }
@article {pmid36696267, year = {2023}, author = {Szewczyk, B and Günther, R and Japtok, J and Frech, MJ and Naumann, M and Lee, HO and Hermann, A}, title = {FUS ALS neurons activate major stress pathways and reduce translation as an early protective mechanism against neurodegeneration.}, journal = {Cell reports}, volume = {42}, number = {2}, pages = {112025}, doi = {10.1016/j.celrep.2023.112025}, pmid = {36696267}, issn = {2211-1247}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/pathology ; *Induced Pluripotent Stem Cells/metabolism ; Motor Neurons/metabolism ; Mutation ; Cytoplasm/metabolism ; RNA-Binding Protein FUS/genetics/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder causing progressive loss of motor neurons. Mutations in Fused in sarcoma (FUS) leading to its cytoplasmic mislocalization cause a subset of ALS. Under stress, mutant FUS localizes to stress granules (SGs)-cytoplasmic condensates composed of RNA and various proteins. Aberrant dynamics of SGs is linked to the pathology of ALS. Here, using motor neurons (MNs) derived from human induced pluripotent stem cells, we show that, in mutant FUS, MN dynamics of SGs is disturbed. Additionally, heat-shock response (HSR) and integrated stress response (ISR) involved in the regulation of SGs are upregulated in mutant MNs. HSR activation correlates with the amount of cytoplasmic FUS mislocalization. While inhibition of SG formation, translation, or ISR does not influence survival of FUS ALS neurons, proteotoxicity that cannot be compensated with the activation of stress pathways is the main driver of neurodegeneration in early FUS ALS.}, }
@article {pmid36701233, year = {2023}, author = {Gallagher, ER and Holzbaur, ELF}, title = {The selective autophagy adaptor p62/SQSTM1 forms phase condensates regulated by HSP27 that facilitate the clearance of damaged lysosomes via lysophagy.}, journal = {Cell reports}, volume = {42}, number = {2}, pages = {112037}, pmid = {36701233}, issn = {2211-1247}, support = {F31 NS125954/NS/NINDS NIH HHS/United States ; R01 NS060698/NS/NINDS NIH HHS/United States ; T32 GM007229/GM/NIGMS NIH HHS/United States ; }, mesh = {Humans ; Amyotrophic Lateral Sclerosis/genetics/metabolism ; Autophagy ; HeLa Cells ; HSP27 Heat-Shock Proteins/genetics/metabolism ; *Lysosomes/metabolism ; *Macroautophagy ; Sequestosome-1 Protein/genetics/metabolism ; }, abstract = {In response to lysosomal damage, cells engage several quality-control mechanisms, including the selective isolation and degradation of damaged lysosomes by lysophagy. Here, we report that the selective autophagy adaptor SQSTM1/p62 is recruited to damaged lysosomes in both HeLa cells and neurons and is required for lysophagic flux. The Phox and Bem1p (PB1) domain of p62 mediates oligomerization and is specifically required for lysophagy. Consistent with this observation, we find that p62 forms condensates on damaged lysosomes. These condensates are precisely tuned by the small heat shock protein HSP27, which is phosphorylated in response to lysosomal injury and maintains the liquidity of p62 condensates, facilitating autophagosome formation. Mutations in p62 have been identified in patients with amyotrophic lateral sclerosis (ALS); ALS-associated mutations in p62 impair lysophagy, suggesting that deficits in this pathway may contribute to neurodegeneration. Thus, p62 condensates regulated by HSP27 promote lysophagy by forming platforms for autophagosome biogenesis at damaged lysosomes.}, }
@article {pmid36789492, year = {2023}, author = {Rafiee, MR and Rohban, S and Davey, K and Ule, J and Luscombe, NM}, title = {RNA polymerase II-associated proteins reveal pathways affected in VCP-related amyotrophic lateral sclerosis.}, journal = {Brain : a journal of neurology}, volume = {146}, number = {6}, pages = {2547-2556}, pmid = {36789492}, issn = {1460-2156}, support = {FC001110/CRUK_/Cancer Research UK/United Kingdom ; }, mesh = {Humans ; Valosin Containing Protein/genetics/metabolism ; *Amyotrophic Lateral Sclerosis/genetics/metabolism ; RNA Polymerase II/metabolism ; Adenosine Triphosphatases/genetics/metabolism ; Mutation/genetics ; Antigens, Neoplasm ; RNA-Binding Proteins/genetics ; Nerve Tissue Proteins/genetics ; }, abstract = {Valosin-containing protein (VCP) is a hexameric ATPase associated with diverse cellular activities. Genetic mutations in VCP are associated with several forms of muscular and neuronal degeneration, including amyotrophic lateral sclerosis (ALS). Moreover, VCP mediates UV-induced proteolysis of RNA polymerase II (RNAPII), but little is known about the effects of VCP mutations on the transcriptional machinery. Here, we used silica particle-assisted chromatin enrichment and mass spectrometry to study proteins co-localized with RNAPII in precursor neurons differentiated from VCP-mutant or control induced pluripotent stem cells. Remarkably, we observed diminished RNAPII binding of proteins involved in transcription elongation and mRNA splicing in mutant cells. One of these is SART3, a recycling factor of the splicing machinery, whose knockdown leads to perturbed intron retention in several ALS-associated genes. Additional reduced proteins are RBM45, EIF5A and RNF220, mutations in which are associated with various neurodegenerative disorders and are linked to TDP-43 aggregation. Conversely, we observed increased RNAPII binding of heat shock proteins such as HSPB1. Together, these findings shed light on how transcription and splicing machinery are impaired by VCP mutations, which might contribute to aberrant alternative splicing and proteinopathy in neurodegeneration.}, }
@article {pmid36865578, year = {2023}, author = {Dixit, A and Chakraborty, A and Nath, JR and Chowdhury, PK and Kundu, B}, title = {Ocular protein optineurin shows reversibility from unfolded states and exhibits chaperone-like activity.}, journal = {RSC advances}, volume = {13}, number = {10}, pages = {6827-6837}, pmid = {36865578}, issn = {2046-2069}, abstract = {Optineurin (OPTN) is a multifunctional, ubiquitously expressed cytoplasmic protein, mutants of which are associated with primary open-angle glaucoma (POAG) and amyotrophic lateral sclerosis (ALS). The most abundant heat shock protein crystallin, known for its remarkable thermodynamic stability and chaperoning activity, allows ocular tissues to withstand stress. The presence of OPTN in ocular tissues is intriguing. Interestingly, OPTN also harbors heat shock elements in its promoter region. Sequence analysis of OPTN exhibits intrinsically disordered regions and nucleic acid binding domains. These properties hinted that OPTN might be endowed with sufficient thermodynamic stability and chaperoning activity. However, these attributes of OPTN have not yet been explored. Here, we studied these properties through thermal and chemical denaturation experiments and monitored the processes using CD, fluorimetry, differential scanning calorimetry, and dynamic light scattering. We found that upon heating, OPTN reversibly forms higher-order multimers. OPTN also displayed a chaperone-like function by reducing the thermal aggregation of bovine carbonic anhydrase. It regains its native secondary structure, RNA-binding property, and melting temperature (T m) after refolding from a thermally as well as chemically denatured state. From our data, we conclude that OPTN, with its unique ability to revert from the stress-mediated unfolded state and its unique chaperoning function, is a valuable protein of the ocular tissues.}, }
@article {pmid36880056, year = {2023}, author = {Tyler, SEB and Tyler, LDK}, title = {Pathways to healing: Plants with therapeutic potential for neurodegenerative diseases.}, journal = {IBRO neuroscience reports}, volume = {14}, number = {}, pages = {210-234}, pmid = {36880056}, issn = {2667-2421}, abstract = {Some of the greatest challenges in medicine are the neurodegenerative diseases (NDs), which remain without a cure and mostly progress to death. A companion study employed a toolkit methodology to document 2001 plant species with ethnomedicinal uses for alleviating pathologies relevant to NDs, focusing on its relevance to Alzheimer's disease (AD). This study aimed to find plants with therapeutic bioactivities for a range of NDs. 1339 of the 2001 plant species were found to have a bioactivity from the literature of therapeutic relevance to NDs such as Parkinson's disease, Huntington's disease, AD, motor neurone diseases, multiple sclerosis, prion diseases, Neimann-Pick disease, glaucoma, Friedreich's ataxia and Batten disease. 43 types of bioactivities were found, such as reducing protein misfolding, neuroinflammation, oxidative stress and cell death, and promoting neurogenesis, mitochondrial biogenesis, autophagy, longevity, and anti-microbial activity. Ethno-led plant selection was more effective than random selection of plant species. Our findings indicate that ethnomedicinal plants provide a large resource of ND therapeutic potential. The extensive range of bioactivities validate the usefulness of the toolkit methodology in the mining of this data. We found that a number of the documented plants are able to modulate molecular mechanisms underlying various key ND pathologies, revealing a promising and even profound capacity to halt and reverse the processes of neurodegeneration.}, }
@article {pmid36931278, year = {2023}, author = {Lee, S and Jun, YW and Linares, GR and Butler, B and Yuva-Adyemir, Y and Moore, J and Krishnan, G and Ruiz-Juarez, B and Santana, M and Pons, M and Silverman, N and Weng, Z and Ichida, JK and Gao, FB}, title = {Downregulation of Hsp90 and the antimicrobial peptide Mtk suppresses poly(GR)-induced neurotoxicity in C9ORF72-ALS/FTD.}, journal = {Neuron}, volume = {111}, number = {9}, pages = {1381-1390.e6}, pmid = {36931278}, issn = {1097-4199}, support = {S10 OD021580/OD/NIH HHS/United States ; RF1 NS101986/NS/NINDS NIH HHS/United States ; R44 NS097094/NS/NINDS NIH HHS/United States ; U24 HG012343/HG/NHGRI NIH HHS/United States ; R01 NS097850/NS/NINDS NIH HHS/United States ; R37 NS057553/NS/NINDS NIH HHS/United States ; R01 AI060025/AI/NIAID NIH HHS/United States ; R01 NS101986/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; *Amyotrophic Lateral Sclerosis/genetics ; C9orf72 Protein/genetics/metabolism ; Dipeptides/genetics ; DNA Repeat Expansion ; Down-Regulation ; Drosophila/metabolism ; *Frontotemporal Dementia/genetics/metabolism ; Motor Neurons/metabolism ; Disease Models, Animal ; Heat-Shock Proteins ; Drosophila Proteins ; Antimicrobial Cationic Peptides ; }, abstract = {GGGGCC repeat expansion in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Repeat RNAs can be translated into dipeptide repeat proteins, including poly(GR), whose mechanisms of action remain largely unknown. In an RNA-seq analysis of poly(GR) toxicity in Drosophila, we found that several antimicrobial peptide genes, such as metchnikowin (Mtk), and heat shock protein (Hsp) genes are activated. Mtk knockdown in the fly eye or in all neurons suppresses poly(GR) neurotoxicity. These findings suggest a cell-autonomous role of Mtk in neurodegeneration. Hsp90 knockdown partially rescues both poly(GR) toxicity in flies and neurodegeneration in C9ORF72 motor neurons derived from induced pluripotent stem cells (iPSCs). Topoisomerase II (TopoII) regulates poly(GR)-induced upregulation of Hsp90 and Mtk. TopoII knockdown also suppresses poly(GR) toxicity in Drosophila and improves survival of C9ORF72 iPSC-derived motor neurons. These results suggest potential novel therapeutic targets for C9ORF72-ALS/FTD.}, }
@article {pmid37194327, year = {2023}, author = {Gallagher, ER and Holzbaur, ELF}, title = {SQSTM1/P62 promotes lysophagy via formation of liquid-like condensates maintained by HSP27.}, journal = {Autophagy}, volume = {19}, number = {11}, pages = {3029-3030}, pmid = {37194327}, issn = {1554-8635}, support = {F31 NS125954/NS/NINDS NIH HHS/United States ; R01 NS060698/NS/NINDS NIH HHS/United States ; }, mesh = {Humans ; Sequestosome-1 Protein/metabolism ; *HSP27 Heat-Shock Proteins/metabolism ; Macroautophagy ; Autophagy ; *Amyotrophic Lateral Sclerosis/metabolism ; }, abstract = {SQSTM1/p62: Sequestosome-1; HSP27: Heat shock protein 27; LLPS: liquid-liquid phase separation; iPSC: induced pluripotent stem cell; PB1: Phox and Bem1p; FRAP: fluorescence recovery after photo-bleaching; ATG: autophagy-related; ALS: amyotrophic lateral sclerosis.}, }
@article {pmid37473581, year = {2023}, author = {Woo, E and Bredvik, K and Liu, B and Fuchs, TJ and Manfredi, G and Konrad, C}, title = {Machine learning approaches based on fibroblast morphometry do not predict ALS.}, journal = {Neurobiology of aging}, volume = {130}, number = {}, pages = {80-83}, doi = {10.1016/j.neurobiolaging.2023.06.010}, pmid = {37473581}, issn = {1558-1497}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/diagnosis/metabolism ; Biomarkers ; Endoplasmic Reticulum/metabolism ; Machine Learning ; Fibroblasts/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a devastating neuromuscular disease with limited therapeutic options. Biomarkers are needed for early disease detection, clinical trial design, and personalized medicine. Early evidence suggests that specific morphometric features in ALS primary skin fibroblasts may be used as biomarkers; however, this hypothesis has not been rigorously tested in conclusively large fibroblast populations. Here, we imaged ALS-relevant organelles (mitochondria, endoplasmic reticulum, lysosomes) and proteins (TAR DNA-binding protein 43, Ras GTPase-activating protein-binding protein 1, heat-shock protein 60) at baseline and under stress perturbations and tested their predictive power on a total set of 443 human fibroblast lines from ALS and healthy individuals. Machine learning approaches were able to confidently predict stress perturbation states (ROC-AUC ∼0.99) but not disease groups or clinical features (ROC-AUC 0.58-0.64). Our findings indicate that multivariate models using patient-derived fibroblast morphometry can accurately predict different stressors but are insufficient to develop viable ALS biomarkers.}, }
@article {pmid37488847, year = {2024}, author = {Huber, CC and Wang, H}, title = {Pathogenic and therapeutic role of exosomes in neurodegenerative disorders.}, journal = {Neural regeneration research}, volume = {19}, number = {1}, pages = {75-79}, pmid = {37488847}, issn = {1673-5374}, support = {P20 GM103443/GM/NIGMS NIH HHS/United States ; P20 RR016479/RR/NCRR NIH HHS/United States ; RF1 AG072510/AG/NIA NIH HHS/United States ; T32 GM136503/GM/NIGMS NIH HHS/United States ; }, abstract = {Neurodegenerative disorders affect millions of people worldwide, and the prevalence of these disorders is only projected to rise as the number of people over 65 will drastically increase in the coming years. While therapies exist to aid in symptomatic relief, effective treatments that can stop or reverse the progress of each neurodegenerative disease are lacking. Recently, research on the role of extracellular vesicles as disease markers and therapeutics has been intensively studied. Exosomes, 30-150 nm in diameter, are one type of extracellular vesicles facilitating cell-to-cell communication. Exosomes are thought to play a role in disease propagation in a variety of neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis. Accordingly, the exosomes derived from the patients are an invaluable source of disease biomarkers. On the other hand, exosomes, especially those derived from stem cells, could serve as a therapeutic for these disorders, as seen by a rapid increase in clinical trials investigating the therapeutic efficacy of exosomes in different neurological diseases. This review summarizes the pathological burden and therapeutic approach of exosomes in neurodegenerative disorders. We also highlight how heat shock increases the yield of exosomes while still maintaining their therapeutic efficacy. Finally, this review concludes with outstanding questions that remain to be addressed in exosomal research.}, }
@article {pmid37516663, year = {2023}, author = {Ahmed, M and Spicer, C and Harley, J and Taylor, JP and Hanna, M and Patani, R and Greensmith, L}, title = {Amplifying the Heat Shock Response Ameliorates ALS and FTD Pathology in Mouse and Human Models.}, journal = {Molecular neurobiology}, volume = {60}, number = {12}, pages = {6896-6915}, pmid = {37516663}, issn = {1559-1182}, support = {MR/S006591/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Humans ; Animals ; Mice ; *Amyotrophic Lateral Sclerosis/drug therapy/genetics/metabolism ; *Frontotemporal Dementia/drug therapy/genetics/pathology ; Hydroxylamines/therapeutic use ; Heat-Shock Response ; Mutation/genetics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are now known as parts of a disease spectrum with common pathological features and genetic causes. However, as both conditions are clinically heterogeneous, patient groups may be phenotypically similar but pathogenically and genetically variable. Despite numerous clinical trials, there remains no effective therapy for these conditions, which, in part, may be due to challenges of therapy development in a heterogeneous patient population. Disruption to protein homeostasis is a key feature of different forms of ALS and FTD. Targeting the endogenous protein chaperone system, the heat shock response (HSR) may, therefore, be a potential therapeutic approach. We conducted a preclinical study of a known pharmacological amplifier of the HSR, called arimoclomol, in mice with a mutation in valosin-containing protein (VCP) which causes both ALS and FTD in patients. We demonstrate that amplification of the HSR ameliorates the ALS/FTD-like phenotype in the spinal cord and brain of mutant VCP mice and prevents neuronal loss, replicating our earlier findings in the SOD1 mouse model of ALS. Moreover, in human cell models, we demonstrate improvements in pathology upon arimoclomol treatment in mutant VCP patient fibroblasts and iPSC-derived motor neurons. Our findings suggest that targeting of the HSR may have therapeutic potential, not only in non-SOD1 ALS, but also for the treatment of FTD.}, }
@article {pmid37795273, year = {2023}, author = {Venediktov, AA and Bushueva, OY and Kudryavtseva, VA and Kuzmin, EA and Moiseeva, AV and Baldycheva, A and Meglinski, I and Piavchenko, GA}, title = {Closest horizons of Hsp70 engagement to manage neurodegeneration.}, journal = {Frontiers in molecular neuroscience}, volume = {16}, number = {}, pages = {1230436}, pmid = {37795273}, issn = {1662-5099}, abstract = {Our review seeks to elucidate the current state-of-the-art in studies of 70-kilodalton-weighed heat shock proteins (Hsp70) in neurodegenerative diseases (NDs). The family has already been shown to play a crucial role in pathological aggregation for a wide spectrum of brain pathologies. However, a slender boundary between a big body of fundamental data and its implementation has only recently been crossed. Currently, we are witnessing an anticipated advancement in the domain with dozens of studies published every month. In this review, we briefly summarize scattered results regarding the role of Hsp70 in the most common NDs including Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). We also bridge translational studies and clinical trials to portray the output for medical practice. Available options to regulate Hsp70 activity in NDs are outlined, too.}, }
@article {pmid37870677, year = {2024}, author = {Wang, S and Zheng, X and Wei, Q and Lin, J and Yang, T and Xiao, Y and Jiang, Q and Li, C and Shang, H}, title = {Rare DNAJC7 Variants May Play a Minor Role in Chinese Patients with ALS.}, journal = {Molecular neurobiology}, volume = {61}, number = {4}, pages = {2265-2269}, pmid = {37870677}, issn = {1559-1182}, support = {2022ZDZX0023//Sichuan Science and Technology Program/ ; }, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/genetics ; Genetic Association Studies ; Mutation, Missense ; Gene Frequency ; China ; Heat-Shock Proteins/genetics ; Molecular Chaperones ; }, abstract = {DnaJ heat shock protein family member C7 gene (DNAJC7) has been identified as a genetic risk factor for amyotrophic lateral sclerosis (ALS). In our study, we aimed to screen for rare variants in DNAJC7 in a large cohort of Chinese ALS patients, and investigate the genotype-phenotype correlation of DNAJC7 in ALS. Four (0.19%) variants of DNAJC7 with minor allele frequency (MAF) < 0.1% among 2124 patients were identified, including 1 protein-truncating variant and 3 missense variants, all of which were predicted to be damaging. The patients carrying variants of DNAJC7 in our cohort tented to have a limb onset and a relatively slow disease progression. However, burden analysis did not show an enrichment of rare damaging variants in ALS patients compared to controls. Further analysis involving diverse regions and larger sample size is necessary to elucidate the role of DNAJC7 in the pathogenicity of ALS.}, }
@article {pmid37873158, year = {2024}, author = {Alecki, C and Rizwan, J and Le, P and Jacob-Tomas, S and Fernandez-Comaduran, M and Verbrugghe, M and Xu, JSM and Minotti, S and Lynch, J and Biswas, J and Wu, T and Durham, H and Yeo, GW and Vera, M}, title = {Localized synthesis of molecular chaperones sustains neuronal proteostasis.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.10.03.560761}, pmid = {37873158}, issn = {2692-8205}, support = {RF1 MH126719/MH/NIMH NIH HHS/United States ; U41 HG009889/HG/NHGRI NIH HHS/United States ; U24 HG009889/HG/NHGRI NIH HHS/United States ; R01 HG004659/HG/NHGRI NIH HHS/United States ; R01 HG011864/HG/NHGRI NIH HHS/United States ; R01 NS103172/NS/NINDS NIH HHS/United States ; }, abstract = {Neurons are challenged to maintain proteostasis in neuronal projections, particularly with the physiological stress at synapses to support intercellular communication underlying important functions such as memory and movement control. Proteostasis is maintained through regulated protein synthesis and degradation and chaperone-assisted protein folding. Using high-resolution fluorescent microscopy, we discovered that neurons localize a subset of chaperone mRNAs to their dendrites, particularly more proximal regions, and increase this asymmetric localization following proteotoxic stress through microtubule-based transport from the soma. The most abundant chaperone mRNA in dendrites encodes the constitutive heat shock protein 70, HSPA8. Proteotoxic stress in cultured neurons, induced by inhibiting proteasome activity or inducing oxidative stress, enhanced transport of Hspa8 mRNAs to dendrites and the percentage of mRNAs engaged in translation on mono and polyribosomes. Knocking down the ALS-related protein Fused in Sarcoma (FUS) and a dominant mutation in the heterogenous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1) impaired stress-mediated localization of Hspa8 mRNA to dendrites in cultured murine motor neurons and human iPSC-derived neurons, respectively, revealing the importance of these RNA-binding proteins in maintaining proteostasis. These results reveal the increased dendritic localization and translation of the constitutive HSP70 Hspa8 mRNA as a crucial neuronal stress response to uphold proteostasis and prevent neurodegeneration.}, }
@article {pmid38168440, year = {2023}, author = {Ugalde, MV and Alecki, C and Rizwan, J and Le, P and Jacob-Tomas, S and Xu, JM and Minotti, S and Wu, T and Durham, H and Yeo, G}, title = {Localized molecular chaperone synthesis maintains neuronal dendrite proteostasis.}, journal = {Research square}, volume = {}, number = {}, pages = {}, pmid = {38168440}, issn = {2693-5015}, support = {RF1 MH126719/MH/NIMH NIH HHS/United States ; U41 HG009889/HG/NHGRI NIH HHS/United States ; U24 HG009889/HG/NHGRI NIH HHS/United States ; R01 HG004659/HG/NHGRI NIH HHS/United States ; R01 HG011864/HG/NHGRI NIH HHS/United States ; R01 NS103172/NS/NINDS NIH HHS/United States ; }, abstract = {Proteostasis is maintained through regulated protein synthesis and degradation and chaperone-assisted protein folding. However, this is challenging in neuronal projections because of their polarized morphology and constant synaptic proteome remodeling. Using high-resolution fluorescence microscopy, we discovered that neurons localize a subset of chaperone mRNAs to their dendrites and use microtubule-based transport to increase this asymmetric localization following proteotoxic stress. The most abundant dendritic chaperone mRNA encodes a constitutive heat shock protein 70 family member (HSPA8). Proteotoxic stress also enhanced HSPA8 mRNA translation efficiency in dendrites. Stress-mediated HSPA8 mRNA localization to the dendrites was impaired by depleting fused in sarcoma-an amyotrophic lateral sclerosis-related protein-in cultured mouse motor neurons and expressing a pathogenic variant of heterogenous nuclear ribonucleoprotein A2/B1 in neurons derived from human induced pluripotent stem cells. These results reveal a crucial and unexpected neuronal stress response in which RNA-binding proteins increase the dendritic localization of HSPA8 mRNA to maintain proteostasis and prevent neurodegeneration.}, }
@article {pmid38260713, year = {2023}, author = {Geraci, J and Bhargava, R and Qorri, B and Leonchyk, P and Cook, D and Cook, M and Sie, F and Pani, L}, title = {Machine learning hypothesis-generation for patient stratification and target discovery in rare disease: our experience with Open Science in ALS.}, journal = {Frontiers in computational neuroscience}, volume = {17}, number = {}, pages = {1199736}, pmid = {38260713}, issn = {1662-5188}, abstract = {INTRODUCTION: Advances in machine learning (ML) methodologies, combined with multidisciplinary collaborations across biological and physical sciences, has the potential to propel drug discovery and development. Open Science fosters this collaboration by releasing datasets and methods into the public space; however, further education and widespread acceptance and adoption of Open Science approaches are necessary to tackle the plethora of known disease states.<br><br>MOTIVATION: In addition to providing much needed insights into potential therapeutic protein targets, we also aim to demonstrate that small patient datasets have the potential to provide insights that usually require many samples (>5,000). There are many such datasets available and novel advancements in ML can provide valuable insights from these patient datasets.<br><br>PROBLEM STATEMENT: Using a public dataset made available by patient advocacy group AnswerALS and a multidisciplinary Open Science approach with a systems biology augmented ML technology, we aim to validate previously reported drug targets in ALS and provide novel insights about ALS subpopulations and potential drug targets using a unique combination of ML methods and graph theory.<br><br>METHODOLOGY: We use NetraAI to generate hypotheses about specific patient subpopulations, which were then refined and validated through a combination of ML techniques, systems biology methods, and expert input.<br><br>RESULTS: We extracted 8 target classes, each comprising of several genes that shed light into ALS pathophysiology and represent new avenues for treatment. These target classes are broadly categorized as inflammation, epigenetic, heat shock, neuromuscular junction, autophagy, apoptosis, axonal transport, and excitotoxicity. These findings are not mutually exclusive, and instead represent a systematic view of ALS pathophysiology. Based on these findings, we suggest that simultaneous targeting of ALS has the potential to mitigate ALS progression, with the plausibility of maintaining and sustaining an improved quality of life (QoL) for ALS patients. Even further, we identified subpopulations based on disease onset.<br><br>CONCLUSION: In the spirit of Open Science, this work aims to bridge the knowledge gap in ALS pathophysiology to aid in diagnostic, prognostic, and therapeutic strategies and pave the way for the development of personalized treatments tailored to the individual's needs.}, }
@article {pmid38334609, year = {2024}, author = {Noori, L and Saqagandomabadi, V and Di Felice, V and David, S and Caruso Bavisotto, C and Bucchieri, F and Cappello, F and Conway de Macario, E and Macario, AJL and Scalia, F}, title = {Putative Roles and Therapeutic Potential of the Chaperone System in Amyotrophic Lateral Sclerosis and Multiple Sclerosis.}, journal = {Cells}, volume = {13}, number = {3}, pages = {}, pmid = {38334609}, issn = {2073-4409}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/metabolism ; *Multiple Sclerosis/therapy ; Molecular Chaperones/metabolism ; Heat-Shock Proteins/metabolism ; }, abstract = {The putative pathogenic roles and therapeutic potential of the chaperone system (CS) in amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS) are reviewed to provide a bibliographic and conceptual platform for launching research on the diagnostic and therapeutic applications of CS components. Various studies suggest that dysfunction of the CS contributes to the pathogenesis of ALS and MS, and here, we identify some of the implicated CS members. The physiology and pathophysiology of the CS members can be properly understood if they are studied or experimentally or clinically manipulated for diagnostic or therapeutic purposes, bearing in mind that they belong to a physiological system with multiple interacting and dynamic components, widespread throughout the body, intra- and extracellularly. Molecular chaperones, some called heat shock protein (Hsp), are the chief components of the CS, whose canonical functions are cytoprotective. However, abnormal chaperones can be etiopathogenic factors in a wide range of disorders, chaperonopathies, including ALS and MS, according to the data reviewed. Chaperones typically form teams, and these build functional networks to maintain protein homeostasis, the canonical role of the CS. However, members of the CS also display non-canonical functions unrelated to protein homeostasis. Therefore, chaperones and other members of the CS, if abnormal, may disturb not only protein synthesis, maturation, and migration but also other physiological processes. Thus, in elucidating the role of CS components in ALS and MS, one must look at protein homeostasis abnormalities and beyond, following the clues emerging from the works discussed here.}, }
@article {pmid38570009, year = {2024}, author = {Fernández Comaduran, M and Minotti, S and Jacob-Tomas, S and Rizwan, J and Larochelle, N and Robitaille, R and Sephton, CF and Vera, M and Nalbantoglu, JN and Durham, HD}, title = {Impact of histone deacetylase inhibition and arimoclomol on heat shock protein expression and disease biomarkers in primary culture models of familial ALS.}, journal = {Cell stress &amp; chaperones}, volume = {29}, number = {3}, pages = {359-380}, pmid = {38570009}, issn = {1466-1268}, mesh = {*Amyotrophic Lateral Sclerosis/metabolism/genetics/drug therapy ; *Histone Deacetylase Inhibitors/pharmacology ; *Biomarkers/metabolism ; *DNA-Binding Proteins/metabolism/genetics ; Humans ; *Motor Neurons/metabolism/drug effects/pathology ; Animals ; HSP70 Heat-Shock Proteins/metabolism/genetics ; HSC70 Heat-Shock Proteins/metabolism/genetics ; Hydroxylamines/pharmacology ; Cells, Cultured ; RNA-Binding Protein FUS/metabolism/genetics ; Superoxide Dismutase-1/metabolism/genetics ; }, abstract = {Protein misfolding and mislocalization are common themes in neurodegenerative disorders, including motor neuron disease, and amyotrophic lateral sclerosis (ALS). Maintaining proteostasis is a crosscutting therapeutic target, including the upregulation of heat shock proteins (HSP) to increase chaperoning capacity. Motor neurons have a high threshold for upregulating stress-inducible HSPA1A, but constitutively express high levels of HSPA8. This study compared the expression of these HSPs in cultured motor neurons expressing three variants linked to familial ALS: TAR DNA binding protein 43 kDa (TDP-43)[G348C], fused in sarcoma (FUS)[R521G], or superoxide dismutase I (SOD1)[G93A]. All variants were poor inducers of Hspa1a, and reduced levels of Hspa8 mRNA and protein, indicating multiple compromises in chaperoning capacity. To promote HSP expression, cultures were treated with the putative HSP coinducer, arimoclomol, and class I histone deacetylase inhibitors, to promote active chromatin for transcription, and with the combination. Treatments had variable, often different effects on the expression of Hspa1a and Hspa8, depending on the ALS variant expressed, mRNA distribution (somata and dendrites), and biomarker of toxicity measured (histone acetylation, maintaining nuclear TDP-43 and the neuronal Brm/Brg-associated factor chromatin remodeling complex component Brg1, mitochondrial transport, FUS aggregation). Overall, histone deacetylase inhibition alone was effective on more measures than arimoclomol. As in the FUS model, arimoclomol failed to induce HSPA1A or preserve Hspa8 mRNA in the TDP-43 model, despite preserving nuclear TDP-43 and Brg1, indicating neuroprotective properties other than HSP induction. The data speak to the complexity of drug mechanisms against multiple biomarkers of ALS pathogenesis, as well as to the importance of HSPA8 for neuronal proteostasis in both somata and dendrites.}, }
@article {pmid38625841, year = {2025}, author = {Savant, R and Pradhan, RK and Bhagat, S and Mythri, RB and Varghese, AM and Vengalil, S and Nalini, A and Sathyaprabha, TN and Raju, TR and Vijayalakshmi, K}, title = {Enhanced levels of fractalkine and HSP60 in cerebrospinal fluid of sporadic amyotrophic lateral sclerosis patients.}, journal = {The International journal of neuroscience}, volume = {135}, number = {9}, pages = {1048-1058}, doi = {10.1080/00207454.2024.2344581}, pmid = {38625841}, issn = {1563-5279}, mesh = {Humans ; *Chaperonin 60/cerebrospinal fluid ; *Chemokine CX3CL1/cerebrospinal fluid ; *Amyotrophic Lateral Sclerosis/cerebrospinal fluid ; Middle Aged ; Female ; Male ; Aged ; Adult ; Biomarkers/cerebrospinal fluid ; Hexosaminidases/cerebrospinal fluid ; *Mitochondrial Proteins/cerebrospinal fluid ; Severity of Illness Index ; }, abstract = {Amyotrophic Lateral Sclerosis (ALS) is a multifactorial neurodegenerative disorder with a significant contribution of non-cell autonomous mechanisms to motor neuronal degeneration. Amongst a plethora of molecules, fractalkine (C-X3-C motif chemokine ligand 1), and Heat Shock Protein 60 (HSP60), are key modulators of microglial activation. The contribution of these molecules in Sporadic ALS (SALS) remains unexplored. To investigate this, fractalkine levels were estimated in Cerebrospinal fluid (CSF) of SALS patients (ALS-CSF; n = 44) by Enzyme-linked Immunosorbent Assay (ELISA) and correlated with clinical parameters including disease severity and duration. CSF HSP60 levels were estimated by Western blotting (ALS-CSF; n = 19). Also, CSF levels of Chitotriosidase-1 (CHIT-1), a microglia-specific neuroinflammatory molecule, were measured and its association, if any, with fractalkine and HSP60 was investigated. Both fractalkine and HSP60 levels were significantly elevated in ALS-CSF. Similar to our earlier observation, CHIT-1 levels were also upregulated. Fractalkine showed a moderate negative correlation with the ALS-Functional Rating Scale (ALSFRS) score indicating its significant rise in mild cases which plateaued in cases with high disease severity. However, no obvious correlation was found between fractalkine, HSP60, and CHIT-1. Our study hints that high fractalkine levels in mild cases might be conferring neuroprotection by combating microglial activation and highlights its importance as a novel therapeutic target for SALS. On the other hand, significantly enhanced levels of HSP60, a pro-inflammatory molecule, hint towards its role in accentuating microgliosis, although, it doesn't act synergistically with CHIT-1. Our study suggests that fractalkine and HSP60 act independently of CHIT-1 to suppress and accentuate neuroinflammation, respectively.}, }
@article {pmid38708921, year = {2024}, author = {Monteiro, KLC and Dos Santos Alcântara, MG and de Aquino, TM and da Silva-Júnior, EF}, title = {Insights on Natural Products Against Amyotrophic Lateral Sclerosis (ALS).}, journal = {Current neuropharmacology}, volume = {22}, number = {7}, pages = {1169-1188}, pmid = {38708921}, issn = {1875-6190}, mesh = {*Amyotrophic Lateral Sclerosis/drug therapy ; Humans ; *Biological Products/therapeutic use/pharmacology ; Animals ; Neuroprotective Agents/therapeutic use/pharmacology ; Plants, Medicinal/chemistry ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that causes the death of motor neurons and consequent muscle paralysis. Despite many efforts to address it, current therapy targeting ALS remains limited, increasing the interest in complementary therapies. Over the years, several herbal preparations and medicinal plants have been studied to prevent and treat this disease, which has received remarkable attention due to their blood-brain barrier penetration properties and low toxicity. Thus, this review presents the therapeutic potential of a variety of medicinal herbs and their relationship with ALS and their physiopathological pathways.}, }
@article {pmid38782015, year = {2024}, author = {Benatar, M and Hansen, T and Rom, D and Geist, MA and Blaettler, T and Camu, W and Kuzma-Kozakiewicz, M and van den Berg, LH and Morales, RJ and Chio, A and Andersen, PM and Pradat, PF and Lange, D and Van Damme, P and Mora, G and Grudniak, M and Elliott, M and Petri, S and Olney, N and Ladha, S and Goyal, NA and Meyer, T and Hanna, MG and Quinn, C and Genge, A and Zinman, L and Jabari, D and Shoesmith, C and Ludolph, AC and Neuwirth, C and Nations, S and Shefner, JM and Turner, MR and Wuu, J and Bennett, R and Dang, H and Sundgreen, C and , }, title = {Safety and efficacy of arimoclomol in patients with early amyotrophic lateral sclerosis (ORARIALS-01): a randomised, double-blind, placebo-controlled, multicentre, phase 3 trial.}, journal = {The Lancet. Neurology}, volume = {23}, number = {7}, pages = {687-699}, doi = {10.1016/S1474-4422(24)00134-0}, pmid = {38782015}, issn = {1474-4465}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/drug therapy ; Male ; Female ; Double-Blind Method ; Middle Aged ; Aged ; *Neuroprotective Agents/therapeutic use/adverse effects ; Treatment Outcome ; Adult ; Hydroxylamines/therapeutic use/adverse effects/pharmacology ; Oxadiazoles/therapeutic use/adverse effects ; }, abstract = {BACKGROUND: Amyotrophic lateral sclerosis is a progressive neurodegenerative disorder leading to muscle weakness and respiratory failure. Arimoclomol, a heat-shock protein-70 (HSP70) co-inducer, is neuroprotective in animal models of amyotrophic lateral sclerosis, with multiple mechanisms of action, including clearance of protein aggregates, a pathological hallmark of sporadic and familial amyotrophic lateral sclerosis. We aimed to evaluate the safety and efficacy of arimoclomol in patients with amyotrophic lateral sclerosis.<br><br>METHODS: ORARIALS-01 was a multinational, randomised, double-blind, placebo-controlled, parallel-group trial done at 29 centres in 12 countries in Europe and North America. Patients were eligible if they were aged 18 years or older and met El Escorial criteria for clinically possible, probable, probable laboratory-supported, definite, or familial amyotrophic lateral sclerosis; had an ALS Functional Rating Scale-Revised score of 35 or more; and had slow vital capacity at 70% or more of the value predicted on the basis of the participant's age, height, and sex. Patients were randomly assigned (2:1) in blocks of 6, stratified by use of a stable dose of riluzole or no riluzole use, to receive oral arimoclomol citrate 1200 mg/day (400 mg three times per day) or placebo. The Randomisation sequence was computer generated centrally. Investigators, study personnel, and study participants were masked to treatment allocation. The primary outcome was the Combined Assessment of Function and Survival (CAFS) rank score over 76 weeks of treatment. The primary outcome and safety were analysed in the modified intention-to-treat population. This trial is registered with ClinicalTrials.gov, NCT03491462, and is completed.<br><br>FINDINGS: Between July 31, 2018, and July 17, 2019, 287 patients were screened, 245 of whom were enrolled in the trial and randomly assigned. The modified intention-to-treat population comprised 239 patients (160 in the arimoclomol group and 79 in the placebo group): 151 (63%) were male and 88 (37%) were female; mean age was 57&#xb7;6 years (SD 10&#xb7;9). CAFS score over 76 weeks did not differ between groups (mean 0&#xb7;51 [SD 0&#xb7;29] in the arimoclomol group vs 0&#xb7;49 [0&#xb7;28] in the placebo group; p=0&#xb7;62). Cliff's delta comparing the two groups was 0&#xb7;039 (95% CI -0&#xb7;116 to 0&#xb7;194). Proportions of participants who died were similar between the treatment groups: 29 (18%) of 160 patients in the arimoclomol group and 18 (23%) of 79 patients in the placebo group. Most deaths were due to disease progression. The most common adverse events were gastrointestinal. Adverse events were more often deemed treatment-related in the arimoclomol group (104 [65%]) than in the placebo group (41 [52%]) and more often led to treatment discontinuation in the arimoclomol group (26 [16%]) than in the placebo group (four [5%]).<br><br>INTERPRETATION: Arimoclomol did not improve efficacy outcomes compared with placebo. Although available biomarker data are insufficient to preclude future strategies that target the HSP response, safety data suggest that a higher dose of arimoclomol would not have been tolerated.<br><br>FUNDING: Orphazyme.}, }
@article {pmid38822416, year = {2024}, author = {Yadav, S and Deepika,  and Moar, K and Kumar, A and Khola, N and Pant, A and Kakde, GS and Maurya, PK}, title = {Reconsidering red blood cells as the diagnostic potential for neurodegenerative disorders.}, journal = {Biology of the cell}, volume = {116}, number = {7}, pages = {e2400019}, doi = {10.1111/boc.202400019}, pmid = {38822416}, issn = {1768-322X}, support = {//Council of Scientific and Industrial Research (CSIR), Government of India/ ; //University Grant Commission (UGC)/ ; HSCIT-3946//Haryana State Council for Science, Innovation, and Technology/ ; //Central University of Haryana, Mahendergarh/ ; //Indian Council of Medical Research (ICMR), Government of India/ ; }, mesh = {Humans ; *Erythrocytes/metabolism ; *Neurodegenerative Diseases/diagnosis/metabolism/blood ; *Biomarkers/metabolism/blood ; Oxidative Stress ; Animals ; Alzheimer Disease/diagnosis/metabolism/blood ; }, abstract = {BACKGROUND: Red blood cells (RBCs) are usually considered simple cells and transporters of gases to tissues.<br><br>HYPOTHESIS: However, recent research has suggested that RBCs may have diagnostic potential in major neurodegenerative disorders (NDDs).<br><br>RESULTS: This review summarizes the current knowledge on changes in RBC in Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and other NDDs. It discusses the deposition of neuronal proteins like amyloid-&#x3b2;, tau, and &#x3b1;-synuclein, polyamines, changes in the proteins of RBCs like band-3, membrane transporter proteins, heat shock proteins, oxidative stress biomarkers, and altered metabolic pathways in RBCs during neurodegeneration. It also highlights the comparison of RBC diagnostic markers to other in-market diagnoses and discusses the challenges in utilizing RBCs as diagnostic tools, such as the need for standardized protocols and further validation studies.<br><br>SIGNIFICANCE STATEMENT: The evidence suggests that RBCs have diagnostic potential in neurodegenerative disorders, and this study can pave the foundation for further research which may lead to the development of novel diagnostic approaches and treatments.}, }
@article {pmid38826044, year = {2024}, author = {Rooney, JPK and Geoghegan, G and O'Reilly, F and Heverin, M and Bose-O'Reilly, S and Casale, F and Chio, A and Günther, K and Schuster, J and Klopstock, T and Ludolph, A and Hardiman, O and Rakete, S}, title = {Serum heat shock protein concentrations are not associated with amyotrophic lateral sclerosis risk or survival in three European populations.}, journal = {Amyotrophic lateral sclerosis &amp; frontotemporal degeneration}, volume = {25}, number = {7-8}, pages = {751-759}, doi = {10.1080/21678421.2024.2358805}, pmid = {38826044}, issn = {2167-9223}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/blood/mortality/epidemiology/diagnosis ; Female ; Male ; Middle Aged ; Aged ; *Biomarkers/blood ; Germany/epidemiology ; HSP70 Heat-Shock Proteins/blood ; Ireland/epidemiology ; Italy/epidemiology ; Heat-Shock Proteins/blood ; HSP90 Heat-Shock Proteins/blood ; Cohort Studies ; Adult ; Europe/epidemiology ; }, abstract = {Introduction: Serum heat shock protein (HSP) concentrations have been reported as potential biomarkers for amyotrophic lateral sclerosis (ALS). Here, we investigate the role of serum HSP70, HSP90, and DNAJC7 as biomarkers for ALS. Methods: Serum samples were collected from ALS patients and volunteer controls from three different clinical cohorts (in Germany, Ireland, and Italy). Serum HSP concentrations were determined using enzyme-linked immunosorbent assay. Descriptive statistics, generalized logistic regression, and Cox proportional hazards models were used to model associations between log serum HSP concentrations and ALS risk. Results: In total, 251 ALS patients and 184 healthy volunteers were included. Logistic regression models failed to find associations between ALS risk and log serum concentration of HSP70 (OR 0.43, 95% CI: 0.10-1.78, p = 0.242), HSP90 (OR 0.95, 95% CI: 0.39-2.37, p = 0.904), or DNAJC7 (OR 1.55, 95% CI: 0.90-2.68, p = 0.118). Survival of ALS patients was not associated with log serum concentration of HSP HSP70 (HR1.06, 95% CI: 0.36-3.14, p = 0.916), HSP90 (HR 1.17, 95% CI: 0.67-2.02, p = 0.584), or DNAJC7 (HR 0.83, 95% CI: 0.57-1.21, p = 0.337). Discussion: We did not replicate previous findings that serum HSP70 and HSP90 concentrations were associated with risk of ALS. DNAJC7 was not associated with ALS risk, and there were no obvious longitudinal patterns in log serum concentrations of HSP70, HSP90, or DNAJC7. In addition, serum HSP concentrations were not associated with ALS survival.}, }
@article {pmid38849544, year = {2024}, author = {Sun, H and Yang, B and Li, Q and Zhu, X and Song, E and Liu, C and Song, Y and Jiang, G}, title = {Polystyrene nanoparticles trigger aberrant condensation of TDP-43 and amyotrophic lateral sclerosis-like symptoms.}, journal = {Nature nanotechnology}, volume = {19}, number = {9}, pages = {1354-1365}, pmid = {38849544}, issn = {1748-3395}, support = {22176206, 22174116 and 22241604//National Natural Science Foundation of China (National Science Foundation of China)/ ; }, mesh = {*Polystyrenes/chemistry/toxicity ; *Amyotrophic Lateral Sclerosis/metabolism/chemically induced ; *DNA-Binding Proteins/metabolism ; Humans ; *Nanoparticles/chemistry ; Animals ; Mice ; Oxidative Stress/drug effects ; Motor Neurons/metabolism/drug effects/pathology ; HSP70 Heat-Shock Proteins/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the dysfunction and progressive death of cerebral and spinal motor neurons. Preliminary epidemiological research has hinted at a relationship between environmental risks and the escalation of ALS, but the underlying reasons remain mostly mysterious. Here we show that nanosize polystyrene plastics (PS) induce ALS-like symptoms and illustrate the related molecular mechanism. When exposed to PS, cells endure internal oxidative stress, which leads to the aggregation of TAR DNA-binding protein 43 kDa (TDP-43), triggering ALS-like characteristics. In addition, the oxidized heat shock protein 70 fails to escort TDP-43 back to the nucleus. The cytoplasmic accumulation of TDP-43 facilitates the formation of a complex between PS and TDP-43, enhancing the condensation and solidification of TDP-43. These findings are corroborated through in silico and in vivo assays. Altogether, our work illustrates a unique toxicological mechanism induced by nanoparticles and provides insights into the connection between environmental pollution and neurodegenerative disorders.}, }
@article {pmid38903116, year = {2024}, author = {Matera, AG and Steiner, RE and Mills, CA and Herring, LE and Garcia, EL}, title = {Chaperoning the chaperones: Proteomic analysis of the SMN complex reveals conserved and etiologic connections to the proteostasis network.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38903116}, issn = {2692-8205}, support = {P30 CA016086/CA/NCI NIH HHS/United States ; R35 GM136435/GM/NIGMS NIH HHS/United States ; }, abstract = {Molecular chaperones and co-chaperones are highly conserved cellular components that perform variety of duties related to the proper three-dimensional folding of the proteome. The web of factors that carries out this essential task is called the proteostasis network (PN). Ribonucleoproteins (RNPs) represent an underexplored area in terms of the connections they make with the PN. The Survival Motor Neuron (SMN) complex is an RNP assembly chaperone and serves as a paradigm for studying how specific small nuclear (sn)RNAs are identified and paired with their client substrate proteins. SMN protein is the eponymous component of a large complex required for the biogenesis of uridine-rich small nuclear ribonucleoproteins (U-snRNPs) and localizes to distinct membraneless organelles in both the nucleus and cytoplasm of animal cells. SMN forms the oligomeric core of this complex, and missense mutations in its YG box self-interaction domain are known to cause Spinal Muscular Atrophy (SMA). The basic framework for understanding how snRNAs are assembled into U-snRNPs is known, the pathways and mechanisms used by cells to regulate their biogenesis are poorly understood. Given the importance of these processes to normal development as well as neurodegenerative disease, we set out to identify and characterize novel SMN binding partners. Here, we carried out affinity purification mass spectrometry (AP-MS) of SMN using stable fly lines exclusively expressing either wildtype or SMA-causing missense alleles. Bioinformatic analyses of the pulldown data, along with comparisons to proximity labeling studies carried out in human cells, revealed conserved connections to at least two other major chaperone systems including heat shock folding chaperones (HSPs) and histone/nucleosome assembly chaperones. Notably, we found that heat shock cognate protein Hsc70-4 and other HspA family members preferentially interacted with SMA-causing alleles of SMN. Hsc70-4 is particularly interesting because its mRNA is aberrantly sequestered by a mutant form of TDP-43 in mouse and Drosophila ALS (Amyotrophic Lateral Sclerosis) disease models. Most important, a missense allele of Hsc70-4 (HspA8 in mammals) was recently identified as a bypass suppressor of the SMA phenotype in mice. Collectively, these findings suggest that chaperone-related dysfunction lies at the etiological root of both ALS and SMA.}, }
@article {pmid38972779, year = {2024}, author = {Pelaez, MC and Fiore, F and Larochelle, N and Dabbaghizadeh, A and Comaduran, MF and Arbour, D and Minotti, S and Marcadet, L and Semaan, M and Robitaille, R and Nalbantoglu, JN and Sephton, CF and Durham, HD}, title = {Reversal of cognitive deficits in FUS[R521G] amyotrophic lateral sclerosis mice by arimoclomol and a class I histone deacetylase inhibitor independent of heat shock protein induction.}, journal = {Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics}, volume = {21}, number = {5}, pages = {e00388}, pmid = {38972779}, issn = {1878-7479}, mesh = {Animals ; *Amyotrophic Lateral Sclerosis/drug therapy/genetics/metabolism ; *Histone Deacetylase Inhibitors/pharmacology/therapeutic use ; Mice ; *Mice, Transgenic ; Heat-Shock Proteins/genetics/metabolism ; Hydroxylamines/pharmacology/therapeutic use ; Cognitive Dysfunction/drug therapy/metabolism ; Disease Models, Animal ; Spinal Cord/drug effects/metabolism ; Humans ; Mice, Inbred C57BL ; }, abstract = {Protein misfolding and mislocalization are common to both familial and sporadic forms of amyotrophic lateral sclerosis (ALS). Maintaining proteostasis through induction of heat shock proteins (HSP) to increase chaperoning capacity is a rational therapeutic strategy in the treatment of ALS. However, the threshold for upregulating stress-inducible HSPs remains high in neurons, presenting a therapeutic obstacle. This study used mouse models expressing the ALS variants FUS[R521G] or SOD1[G93A] to follow up on previous work in cultured motor neurons showing varied effects of the HSP co-inducer, arimoclomol, and class I histone deacetylase (HDAC) inhibitors on HSP expression depending on the ALS variant being expressed. As in cultured neurons, neither expression of the transgene nor drug treatments induced expression of HSPs in cortex, spinal cord or muscle of FUS[R521G] mice, indicating suppression of the heat shock response. Nonetheless, arimoclomol, and RGFP963, restored performance on cognitive tests and improved cortical dendritic spine densities. In SOD1[G93A] mice, multiple HSPs were upregulated in hindlimb skeletal muscle, but not in lumbar spinal cord with the exception of HSPB1 associated with astrocytosis. Drug treatments improved contractile force but reduced the increase in HSPs in muscle rather than facilitating their expression. The data point to mechanisms other than amplification of the heat shock response underlying recovery of cognitive function in ALS-FUS mice by arimoclomol and class I HDAC inhibition and suggest potential benefits in counteracting cognitive impairment in ALS, frontotemporal dementia and related disorders.}, }
@article {pmid38979013, year = {2024}, author = {Shi, W and Ding, R and Chen, Y and Ji, F and Ji, J and Ma, W and Jin, J}, title = {The HRD1-SEL1L ubiquitin ligase regulates stress granule homeostasis in couple with distinctive signaling branches of ER stress.}, journal = {iScience}, volume = {27}, number = {7}, pages = {110196}, pmid = {38979013}, issn = {2589-0042}, abstract = {Stress granules (SGs) are membrane-less cellular compartments which are dynamically assembled via biomolecular condensation mechanism when eukaryotic cells encounter environmental stresses. SGs are important for gene expression and cell fate regulation. Dysregulation of SG homeostasis has been linked to human neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here we report that the HRD1-SEL1L ubiquitin ligase complex specifically regulates the homeostasis of heat shock-induced SGs through the ubiquitin-proteasome system (UPS) and the UPS-associated ATPase p97. Mechanistically, the HRD1-SEL1L complex mediates SG homeostasis through the BiP-coupled PERK-eIF2α signaling axis of endoplasmic reticulum (ER) stress, thereby coordinating the unfolded protein response (UPR) with SG dynamics. Furthermore, we show that the distinctive branches of ER stress play differential roles in SG homeostasis. Our study indicates that the UPS and the UPR together via the HRD1-SEL1L ubiquitin ligase to maintain SG homeostasis in a stressor-dependent manner.}, }
@article {pmid39492846, year = {2024}, author = {Matera, AG and Steiner, RE and Mills, CA and McMichael, BD and Herring, LE and Garcia, EL}, title = {Proteomic analysis of the SMN complex reveals conserved and etiologic connections to the proteostasis network.}, journal = {Frontiers in RNA research}, volume = {2}, number = {}, pages = {}, pmid = {39492846}, issn = {2813-7116}, support = {P30 CA016086/CA/NCI NIH HHS/United States ; R35 GM136435/GM/NIGMS NIH HHS/United States ; }, abstract = {INTRODUCTION: Molecular chaperones and co-chaperones are highly conserved cellular components that perform a variety of duties related to the proper three-dimensional folding of the proteome. The web of factors that carries out this essential task is called the proteostasis network (PN). Ribonucleoproteins (RNPs) represent an underexplored area in terms of the connections they make with the PN. The Survival Motor Neuron (SMN) complex is an assembly chaperone and serves as a paradigm for studying how specific RNAs are identified and paired with their client substrate proteins to form RNPs. SMN is the eponymous component of a large complex, required for the biogenesis of uridine-rich small nuclear ribonucleoproteins (U-snRNPs), that localizes to distinct membraneless organelles in both the nucleus and cytoplasm of animal cells. SMN protein forms the oligomeric core of this complex, and missense mutations in the human SMN1 gene are known to cause Spinal Muscular Atrophy (SMA). The basic framework for understanding how snRNAs are assembled into U-snRNPs is known. However, the pathways and mechanisms used by cells to regulate their biogenesis are poorly understood.<br><br>METHODS: Given the importance of these processes to normal development as well as neurodegenerative disease, we set out to identify and characterize novel SMN binding partners. We carried out affinity purification mass spectrometry (AP-MS) of Drosophila SMN complexes using fly lines exclusively expressing either wildtype or SMA-causing missense alleles.<br><br>RESULTS: Bioinformatic analyses of the pulldown data, along with comparisons to proximity labeling studies carried out in human cells, revealed conserved connections to at least two other major chaperone systems including heat shock folding chaperones (HSPs) and histone/nucleosome assembly chaperones. Notably, we found that heat shock cognate protein Hsc70-4 and other HspA family members preferentially associated with SMA-causing alleles of SMN.<br><br>DISCUSSION: Hsc70-4 is particularly interesting because its mRNA is aberrantly sequestered by a mutant form of TDP-43 in mouse and Drosophila ALS (Amyotrophic Lateral Sclerosis) disease models. Most important, a missense allele of Hsc70-4 (HspA8 in mammals) was recently identified as a bypass suppressor of the SMA phenotype in mice. Collectively, these findings suggest that chaperone-related dysfunction lies at the etiological root of both ALS and SMA.}, }
@article {pmid39651147, year = {2024}, author = {Fleming, AC and Rao, NR and Wright, M and Savas, JN and Kiskinis, E}, title = {The ALS-associated co-chaperone DNAJC7 mediates neuroprotection against proteotoxic stress by modulating HSF1 activity.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {39651147}, issn = {2692-8205}, support = {F31 NS132580/NS/NINDS NIH HHS/United States ; R01 NS104219/NS/NINDS NIH HHS/United States ; R21 NS131713/NS/NINDS NIH HHS/United States ; S10 OD032464/OD/NIH HHS/United States ; }, abstract = {The degeneration of neurons in patients with amyotrophic lateral sclerosis (ALS) is commonly associated with accumulation of misfolded, insoluble proteins. Heat shock proteins (HSPs) are central regulators of protein homeostasis as they fold newly synthesized proteins and refold damaged proteins. Heterozygous loss-of-function mutations in the DNAJC7 gene that encodes an HSP co-chaperone were recently identified as a cause for rare forms of ALS, yet the mechanisms underlying pathogenesis remain unclear. Using mass spectrometry, we found that the DNAJC7 interactome in human motor neurons (MNs) is enriched for RNA binding proteins (RBPs) and stress response chaperones. MNs generated from iPSCs with the ALS-associated mutation R156X in DNAJC7 exhibit increased insolubility of its client RBP HNRNPU and associated RNA metabolism alterations. Additionally, DNAJC7 haploinsufficiency renders MNs increasingly susceptible to proteotoxic stress and cell death as a result of an ablated HSF1 stress response pathway. Critically, expression of HSF1 in mutant DNAJC7 MNs is sufficient to rescue their sensitivity to proteotoxic stress, while postmortem ALS patient cortical neurons exhibit a reduction in the expression of HSF1 pathway genes. Taken together, our work identifies DNAJC7 as a crucial mediator of HNRNPU function and stress response pathways in human MNs and highlights HSF1 as a therapeutic target in ALS.}, }
@article {pmid39724103, year = {2024}, author = {Garcia-Toscano, L and Currey, HN and Hincks, JC and Stair, JG and Lehrbach, NJ and Liachko, NF}, title = {Decreased Hsp90 activity protects against TDP-43 neurotoxicity in a C. elegans model of amyotrophic lateral sclerosis.}, journal = {PLoS genetics}, volume = {20}, number = {12}, pages = {e1011518}, pmid = {39724103}, issn = {1553-7404}, support = {P40 OD010440/OD/NIH HHS/United States ; R01 AG066729/AG/NIA NIH HHS/United States ; I01 BX005762/BX/BLRD VA/United States ; I01 BX004044/BX/BLRD VA/United States ; R35 GM142728/GM/NIGMS NIH HHS/United States ; P30 DK017047/DK/NIDDK NIH HHS/United States ; }, mesh = {Animals ; *Caenorhabditis elegans/genetics/metabolism ; *Amyotrophic Lateral Sclerosis/genetics/metabolism ; *HSP90 Heat-Shock Proteins/metabolism/genetics ; *Disease Models, Animal ; *DNA-Binding Proteins/metabolism/genetics ; *Caenorhabditis elegans Proteins/metabolism/genetics ; Humans ; Phosphorylation ; Mutation ; Heat-Shock Response/genetics ; TDP-43 Proteinopathies/genetics/metabolism ; }, abstract = {Neuronal inclusions of hyperphosphorylated TDP-43 are hallmarks of disease for most patients with amyotrophic lateral sclerosis (ALS). Mutations in TARDBP, the gene coding for TDP-43, can cause some cases of familial inherited ALS (fALS), indicating dysfunction of TDP-43 drives disease. Aggregated, phosphorylated TDP-43 may contribute to disease phenotypes; alternatively, TDP-43 aggregation may be a protective cellular response sequestering toxic protein away from the rest of the cell. The heat shock responsive chaperone Hsp90 has been shown to interact with TDP-43 and stabilize its normal conformation; however, it is not known whether this interaction contributes to neurotoxicity in vivo. Using a C. elegans model of fALS mutant TDP-43 proteinopathy, we find that loss of function of HSP-90 protects against TDP-43 neurotoxicity and subsequent neurodegeneration in adult animals. This protection is accompanied by a decrease in both total and phosphorylated TDP-43 protein. We also find that hsp-90 mutation or inhibition upregulates key stress responsive heat shock pathway gene expression, including hsp-70 and hsp-16.1, and we demonstrate that normal levels of hsp-16.1 are required for hsp-90 mutation effects on TDP-43. We also observe that the neuroprotective effect due to HSP-90 dysfunction does not involve direct regulation of proteasome activity in C. elegans. Our data demonstrate for the first time that Hsp90 chaperone activity contributes to adverse outcomes in TDP-43 proteinopathies in vivo using a whole animal model of ALS.}, }
@article {pmid39737952, year = {2024}, author = {Alecki, C and Rizwan, J and Le, P and Jacob-Tomas, S and Comaduran, MF and Verbrugghe, M and Xu, JMS and Minotti, S and Lynch, J and Biswas, J and Wu, T and Durham, HD and Yeo, GW and Vera, M}, title = {Localized molecular chaperone synthesis maintains neuronal dendrite proteostasis.}, journal = {Nature communications}, volume = {15}, number = {1}, pages = {10796}, pmid = {39737952}, issn = {2041-1723}, support = {RF1 MH126719/MH/NIMH NIH HHS/United States ; 300232//Fonds de Recherche du Qu&#xe9;bec - Sant&#xe9; (Fonds de la recherche en sante du Quebec)/ ; MH126719//U.S. Department of Health &amp; Human Services | National Institutes of Health (NIH)/ ; HG009889//U.S. Department of Health &amp; Human Services | National Institutes of Health (NIH)/ ; U24 HG009889/HG/NHGRI NIH HHS/United States ; R01 HG004659/HG/NHGRI NIH HHS/United States ; HG011864//U.S. Department of Health &amp; Human Services | National Institutes of Health (NIH)/ ; PJT-186141//Gouvernement du Canada | Canadian Institutes of Health Research (Instituts de Recherche en Sant&#xe9; du Canada)/ ; HG004659//U.S. Department of Health &amp; Human Services | National Institutes of Health (NIH)/ ; Hudson Translational Team Grant//ALS Society of Canada (ALS Canada)/ ; R01 HG011864/HG/NHGRI NIH HHS/United States ; R01 NS103172/NS/NINDS NIH HHS/United States ; 2022-ALS Discovery Grant//ALS Society of Canada (ALS Canada)/ ; P30 CA008748/CA/NCI NIH HHS/United States ; U41 HG009889/HG/NHGRI NIH HHS/United States ; }, mesh = {*Dendrites/metabolism ; *Proteostasis ; Animals ; Humans ; Mice ; *RNA-Binding Protein FUS/metabolism/genetics ; *RNA, Messenger/metabolism/genetics ; *Motor Neurons/metabolism ; *HSC70 Heat-Shock Proteins/metabolism/genetics ; *Hippocampus/metabolism/cytology ; Spinal Cord/metabolism ; Induced Pluripotent Stem Cells/metabolism ; Protein Biosynthesis ; Molecular Chaperones/metabolism/genetics ; HSP70 Heat-Shock Proteins/metabolism/genetics ; Microtubules/metabolism ; }, abstract = {Proteostasis is maintained through regulated protein synthesis and degradation and chaperone-assisted protein folding. However, this is challenging in neuronal projections because of their polarized morphology and constant synaptic proteome remodeling. Using high-resolution fluorescence microscopy, we discover that hippocampal and spinal cord motor neurons of mouse and human origin localize a subset of chaperone mRNAs to their dendrites and use microtubule-based transport to increase this asymmetric localization following proteotoxic stress. The most abundant dendritic chaperone mRNA encodes a constitutive heat shock protein 70 family member (HSPA8). Proteotoxic stress also enhances HSPA8 mRNA translation efficiency in dendrites. Stress-mediated HSPA8 mRNA localization to the dendrites is impaired by depleting fused in sarcoma-an amyotrophic lateral sclerosis-related protein-in cultured spinal cord mouse motor neurons or by expressing a pathogenic variant of heterogenous nuclear ribonucleoprotein A2/B1 in neurons derived from human induced pluripotent stem cells. These results reveal a neuronal stress response in which RNA-binding proteins increase the dendritic localization of HSPA8 mRNA to maintain proteostasis and prevent neurodegeneration.}, }
@article {pmid39804774, year = {2025}, author = {Agnihotri, D and Lee, CC and Lu, PC and He, RY and Huang, YA and Kuo, HC and Huang, JJ}, title = {C9ORF72 poly-PR induces TDP-43 nuclear condensation via NEAT1 and is modulated by HSP70 activity.}, journal = {Cell reports}, volume = {44}, number = {1}, pages = {115173}, doi = {10.1016/j.celrep.2024.115173}, pmid = {39804774}, issn = {2211-1247}, mesh = {Humans ; *HSP70 Heat-Shock Proteins/metabolism ; *C9orf72 Protein/metabolism/genetics ; *DNA-Binding Proteins/metabolism ; *Cell Nucleus/metabolism ; *RNA, Long Noncoding/metabolism/genetics ; *Peptides/metabolism ; Amyotrophic Lateral Sclerosis/metabolism/genetics/pathology ; }, abstract = {The toxicity of C9ORF72-encoded polyproline-arginine (poly-PR) dipeptide is associated with its ability to disrupt the liquid-liquid phase separation of intrinsically disordered proteins participating in the formation of membraneless organelles, such as the nucleolus and paraspeckles. Amyotrophic lateral sclerosis (ALS)-related TAR DNA-binding protein 43 (TDP-43) also undergoes phase separation to form nuclear condensates (NCs) in response to stress. However, whether poly-PR alters the nuclear condensation of TDP-43 in ALS remains unclear. In this study, we find that the poly-PR dipeptide enhances the formation of TDP-43 NCs with decreased fluidity. While the non-coding RNA, nuclear-enriched abundant transcript 1 (NEAT1), is essential for the formation of TDP-43 NCs, heat shock protein 70 (HSP70) chaperone maintains their fluidity. Under prolonged poly-PR stress, HSP70 delocalizes from TDP-43 NCs, leading to the oligomerization of TDP-43 within these condensates. This phenomenon is accompanied with TDP-43 mislocalization and increasing cytotoxicity. Our study demonstrates the role of NEAT1 and HSP70 in the aberrant phase transition of TDP-43 NCs under poly-PR stress.}, }
@article {pmid39871559, year = {2025}, author = {Rani, S and Tuteja, M}, title = {Chaperones as Potential Pharmacological Targets for Treating Protein Aggregation Illness.}, journal = {Current protein &amp; peptide science}, volume = {26}, number = {6}, pages = {451-466}, pmid = {39871559}, issn = {1875-5550}, mesh = {Humans ; *Molecular Chaperones/metabolism ; Animals ; Protein Folding/drug effects ; *Neurodegenerative Diseases/metabolism/drug therapy ; Heat-Shock Proteins/metabolism ; *Protein Aggregation, Pathological/drug therapy/metabolism ; Protein Aggregates/drug effects ; Molecular Targeted Therapy ; *Proteostasis Deficiencies/drug therapy/metabolism ; }, abstract = {The three-dimensional structure of proteins, achieved through the folding of the nascent polypeptide chain in vivo, is largely facilitated by molecular chaperones, which are crucial for determining protein functionality. In addition to aiding in the folding process, chaperones target misfolded proteins for degradation, acting as a quality control system within the cell. Defective protein folding has been implicated in a wide range of clinical conditions, including neurodegenerative and metabolic disorders. It is now well understood that the pathogenesis of neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, Amyotrophic Lateral Sclerosis, and Creutzfeldt-Jakob disease shares a common mechanism: the accumulation of misfolded proteins, which aggregate and become toxic to cells. Among the family of molecular chaperones, Heat Shock Proteins (HSPs) are highly expressed in response to cellular stress and play a pivotal role in preventing protein aggregation. Specific chaperones, particularly HSPs, are now recognized as critical in halting the accumulation and aggregation of misfolded proteins in these conditions. Consequently, these chaperones are increasingly considered promising pharmacological targets for the treatment of protein aggregation-related diseases. This review highlights research exploring the potential roles of specific molecular chaperones in disorders characterized by the accumulation of misfolded proteins.}, }
@article {pmid39887552, year = {2025}, author = {Dopler, MB and Abeer, MI and Arezoumandan, S and Cox, K and Petersen, TL and Daniel, EH and Cannon, CL and Bautista, A and Blancher, KD and Bland, AM and Bond, KJ and Davis, DA and Francois, JM and McCray, EJ and Morgan, JM and Pulliam, JL and Robinson, ZA and Taylor, MJ and Dowell, JA and Cairns, NJ and Gitcho, MA}, title = {A cellular model of TDP-43 induces phosphorylated TDP-43 aggregation with distinct changes in solubility and autophagy dysregulation.}, journal = {The FEBS journal}, volume = {292}, number = {18}, pages = {4870-4897}, pmid = {39887552}, issn = {1742-4658}, support = {52008702//HHMI/ ; T32 GM144895/GM/NIGMS NIH HHS/United States ; 2023004//National Science Foundation/ ; P30 GM145765/GM/NIGMS NIH HHS/United States ; P20GM103446/GM/NIGMS NIH HHS/United States ; P20 GM103446/GM/NIGMS NIH HHS/United States ; 0823//The Paul H. Boerger Fund of the Delaware Community Foundation/ ; P20 GM103653/GM/NIGMS NIH HHS/United States ; R25 GM122722/GM/NIGMS NIH HHS/United States ; P20GM103653/GM/NIGMS NIH HHS/United States ; T32GM144895-03/GM/NIGMS NIH HHS/United States ; P20GM103446/GM/NIGMS NIH HHS/United States ; P20GM103653/GM/NIGMS NIH HHS/United States ; T32GM144895-03/GM/NIGMS NIH HHS/United States ; }, mesh = {*Autophagy ; Solubility ; *DNA-Binding Proteins/genetics/metabolism ; Cell Line ; Humans ; Cytoplasm/metabolism ; Cell Nucleus/metabolism ; Mutation ; Glycolysis ; Phosphorylation ; alpha Karyopherins/genetics/metabolism ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Protein Disulfide-Isomerases/genetics/metabolism ; Proteomics ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease that affects neurons in the brain and spinal cord, causing loss of muscle control, and eventually leads to death. Phosphorylated transactive response DNA binding protein-43 (TDP-43) is the major pathological protein in both sporadic and familial ALS, forming cytoplasmic aggregates in over 95% of cases. Of the 10-15% of ALS cases that are familial, mutations in TDP-43 represent about 5% of those with a family history. We have developed an in vitro overexpression model by introducing three familial ALS mutations (A315T, M337V, and S379P) in the TDP-43 (TARDBP) gene which we define as 3X-TDP-43. This overexpression model TDP-43 shows deficits in autophagy flux and colocalization of TDP-43 with stress granules. We also observe a progressive shift of TDP-43 to the cytoplasm in this model. This overexpression model shows a reduction in solubility of phosphorylated TDP-43 from RIPA to urea soluble. Four glycolytic enzymes, phosphoglycerate kinase one (PGK1), aldolase A (ALDOA), enolase 1 (ENO1), and pyruvate dehydrogenase kinase 1 (PDK1) show significant time-dependent decreases in 3X-TDP-43 expressing cells. Shotgun proteomic analysis shows global changes in the importin subunit alpha-1 (KPNA2), heat shock 70 kDa protein 1A (HSPA1A), and protein disulfide-isomerase A3 (PDIA3) expression levels and coimmunoprecipitation reveals that these proteins complex with TDP-43. Overall, these results suggest that the 3X-TDP-43 model may provide new insights into pathophysiology and an avenue for drug screening in vitro for those suffering from ALS and related TDP-43 proteinopathies.}, }
@article {pmid39907139, year = {2025}, author = {Matera, AG}, title = {Chaperone dysfunction in motor neuron disease: new insights from studies of the SMN complex.}, journal = {Genetics}, volume = {229}, number = {3}, pages = {}, pmid = {39907139}, issn = {1943-2631}, support = {R35 GM136435/GM/NIGMS NIH HHS/United States ; //USA National Institutes of Health/ ; }, mesh = {Humans ; Muscular Atrophy, Spinal/genetics/metabolism ; Amyotrophic Lateral Sclerosis/genetics/metabolism ; *SMN Complex Proteins/metabolism/genetics ; *Molecular Chaperones/metabolism/genetics ; Animals ; *Motor Neuron Disease/genetics/metabolism ; Motor Neurons/metabolism ; }, abstract = {Spinal muscular atrophy and amyotrophic lateral sclerosis are devastating neurodegenerative diseases characterized by motor neuron loss. Although these 2 disorders have distinct genetic origins, recent studies suggest that they share common etiological mechanisms rooted in proteostatic dysfunction. At the heart of this emerging understanding is the survival motor neuron (SMN) complex.}, }
@article {pmid39929612, year = {2025}, author = {Chen, BL and Lu, JZ and Zhou, XM and Wen, XD and Jiang, YJ and Luo, N}, title = {[Mechanism of Daotan Xixin Decoction in treating APP/PS1 mice based on high-throughput sequencing technology and bioinformatics analysis].}, journal = {Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica}, volume = {50}, number = {2}, pages = {301-313}, doi = {10.19540/j.cnki.cjcmm.20241011.401}, pmid = {39929612}, issn = {1001-5302}, mesh = {Animals ; *Drugs, Chinese Herbal/administration &amp; dosage ; Mice ; Male ; *Alzheimer Disease/drug therapy/genetics/metabolism ; Computational Biology ; Mice, Inbred C57BL ; High-Throughput Nucleotide Sequencing ; *Amyloid beta-Protein Precursor/genetics/metabolism ; Hippocampus/drug effects/metabolism ; Mice, Transgenic ; *Presenilin-1/genetics/metabolism ; Humans ; Memory/drug effects ; Maze Learning/drug effects ; Amyloid beta-Peptides/metabolism/genetics ; Disease Models, Animal ; }, abstract = {This study aims to investigate the therapeutic effect and mechanism of Daotan Xixin Decoction on APP/PS1 mice. Twelve APP/PS1 male mice were randomized into four groups: APP/PS1 and low-, medium-, and high-dose Daotan Xixin Decoction. Three C57BL/6 wild-type mice were used as the control group. The learning and memory abilities of mice in each group were examined by the Morris water maze test. The pathological changes of hippocampal nerve cells were observed by hematoxylin-eosin staining and Nissl staining. Immunohistochemistry was employed to detect the expression of β-amyloid(Aβ)_(1-42) in the hippocampal tissue. The high-dose Daotan Xixin Decoction group with significant therapeutic effects and the model group were selected for high-throughput sequencing. The differentially expressed gene(DEG) analysis, Gene Ontology(GO) analysis, Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis, and Gene Set Variation Analysis(GSVA) were performed on the sequencing results. RT-qPCR and Western blot were conducted to determine the mRNA and protein levels, respectively, of some DEGs. Compared with the APP/PS1 group, Daotan Xixin Decoction at different doses significantly improved the learning and memory abilities of APP/PS1 mice, ameliorated the neuropathological damage in the CA1 region of the hippocampus, increased the number of neurons, and decreased the deposition of Aβ_(1-42) in the brain. A total of 1 240 DEGs were screened out, including 634 genes with up-regulated expression and 606 genes with down-regulated expression. The GO analysis predicted the biological processes including RNA splicing and protein folding, the cellular components including spliceosome complexes and nuclear spots, and the molecular functions including unfolded protein binding and heat shock protein binding. The KEGG pathway enrichment analysis revealed the involvement of neurodegenerative disease pathways, amyotrophic lateral sclerosis, and splicing complexes. Further GSVA pathway enrichment analysis showed that the down-regulated pathways involved nuclear factor-κB(NF-κB)-mediated tumor necrosis factor-α(TNF-α) signaling pathway, UV response, and unfolded protein response, while the up-regulated pathways involved the Wnt/β-catenin signaling pathway. The results of RT-qPCR and Western blot showed that compared with the APP/PS1 group, Daotan Xixin Decoction at different doses down-regulated the mRNA and protein levels of signal transducer and activator of transcription 3(STAT3), NF-κB, and interleukin-6(IL-6) in the hippocampus. In conclusion, Daotan Xixin Decoction can improve the learning and memory abilities of APP/PS1 mice by regulating the STAT3/NF-κB/IL-6 signaling pathway.}, }
@article {pmid40054065, year = {2025}, author = {Tserennadmid, B and Nam, MK and Park, JH and Rhim, H and Kang, S}, title = {HAP/ClpP-mediated disaggregation and degradation of Mutant SOD1 aggregates: A potential therapeutic strategy for Amyotrophic lateral sclerosis (ALS).}, journal = {Biochemical and biophysical research communications}, volume = {756}, number = {}, pages = {151533}, doi = {10.1016/j.bbrc.2025.151533}, pmid = {40054065}, issn = {1090-2104}, mesh = {*Amyotrophic Lateral Sclerosis/metabolism/genetics/therapy ; *Superoxide Dismutase-1/metabolism/genetics/chemistry ; Humans ; Protein Aggregates ; Proteolysis ; *Heat-Shock Proteins/metabolism/genetics ; Mutation ; Fluorescence Resonance Energy Transfer ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease marked by the accumulation of misfolded Cu/Zn superoxide dismutase (SOD1) protein aggregates in motor neurons, leading to progressive motor dysfunction and ultimately death. While the molecular chaperone heat shock protein 104 (Hsp104) has been shown to reduce protein misfolding by disaggregating protein aggregates, fully degrading these disaggregated proteins remains a significant challenge. In this study, we have investigated the effects of Hsp104 and its hyperactive variant, HAP, in combination with caseinolytic protease P (CIpP), on the disaggregation and degradation of SOD1 aggregates. Using laser confocal microscopy, fluorescence loss in photobleaching (FLIP), and biomolecular fluorescence complementation (BiFC)-fluorescence resonance energy transfer (FRET) assays, we demonstrate that Hsp104 effectively disaggregates SOD1 aggregates across 14 different G93 mutants, classified based on the properties of substituted amino acids, thus restoring protein mobility. Notably, the HAP/CIpP system not only disaggregates ALS-associated SOD1[G93A] aggregates but also promotes their proteolytic degradation, as evidenced by a significant reduction in high-order oligomers observed through BiFC and FRET assays. This dual mechanism of action presents. the HAP/CIpP system holds significant therapeutic potential for ALS and other neurodegenerative diseases characterized by protein aggregates, as it enables both effective disaggregation and degradation of toxic protein aggregates, thereby maintaining protein homeostasis.}, }
@article {pmid40583948, year = {2025}, author = {Uzunçakmak-Uyanık, H and Tan, E and Temuçin, &#xc7;M}, title = {A Case of Distal Hereditary Motor Neuropathy with HSPB1 Mutation in Coexistence with Myotonia and Myopathy.}, journal = {Noro psikiyatri arsivi}, volume = {62}, number = {2}, pages = {205-206}, pmid = {40583948}, issn = {1300-0667}, abstract = {Distal hereditary motor neuropathies (dHMNs), also named as distal spinal muscular atrophy, are a group of disorders that cause degeneration of motor nerves. Currently, only 15% to 32.5% of patients with dHMN have been genetically identified. The most common cause of dHMNs gene mutations is HSPB1 mutation. In HSPB1 mutation, which is also one of the myopathogens via satellite cell pathology, dHMNS may coexist with neuromuscular junction disorder, motor neuron disease, satellite cell dysfunction and therefore myopathic findings. No case of myopathy and myotonia with HSPB1 mutation has been reported in the literature yet. We present a case with electrophysiologic findings in HSPB1 mutation by discussing the possible mechanisms underlying myotonic discharges and myopathic findings.}, }
@article {pmid40622676, year = {2025}, author = {Wei, Y and Li, D and Yang, R and Liu, Y and Luo, X and Zhao, W and Yang, H and Chen, Z and Shen, C and Wang, Y and Huang, Z}, title = {TIA1-mediated stress granules promote neurodegeneration by sequestering HSP70 mRNA in C9orf72 mice.}, journal = {Brain : a journal of neurology}, volume = {148}, number = {12}, pages = {4482-4494}, doi = {10.1093/brain/awaf248}, pmid = {40622676}, issn = {1460-2156}, mesh = {Animals ; *T-Cell Intracellular Antigen-1/metabolism/genetics ; *HSP70 Heat-Shock Proteins/metabolism/genetics ; *Amyotrophic Lateral Sclerosis/metabolism/pathology/genetics ; Mice ; *Stress Granules/metabolism/pathology ; *C9orf72 Protein/genetics/metabolism ; Humans ; Mice, Knockout ; RNA, Messenger/metabolism ; Male ; Motor Neurons/metabolism/pathology ; Motor Cortex/metabolism/pathology ; Disease Models, Animal ; Female ; Nerve Degeneration/pathology/metabolism ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease with progressive loss of motor neurons in the central nervous system. Recent studies have reported that there are mutations at the T-cell antigen-1 (TIA1) domain site in some ALS patients. TIA1 is a key component of stress granules (SGs), but its role and mechanism in ALS pathogenesis remain unclear. In this study, we found that TIA1 was upregulated in the motor cortex of post-mortem ALS patients as well as in the motor cortex neurons of C9orf72-poly-Gly-Ala (GA) mice (ALS mice). TIA1 knockout in the CNS [TIA1Nestin-conditional knockout (CKO) mice] alleviated motor neuron loss, neuroinflammation and motor dysfunction in C9orf72-poly-GA mice. Mechanistically, RNA sequencing combined with the C9orf72-ALS/frontotemporal dementia patient single nucleus RNA-sequencing database revealed that mRNA of heat shock protein 70 (HSP70) family member genes such as HSPa1b were upregulated in the motor cortex of TIA1Nestin-CKO ALS mice. We further found that TIA1-mediated SG formation was increased during ALS pathogenesis, leading to HSP70 mRNA being sequestered into SGs. This reduced HSP70 expression, impairing the degradation of poly-GA aggregates by the UBQLN2-HSP70 pathway and exacerbating C9orf72-ALS progression. Taken together, these findings highlight a previously unrecognized role of TIA1-mediated SGs in promoting ALS pathogenesis by sequestering HSP70 mRNA, suggesting potential therapeutic targets for ALS treatment.}, }
@article {pmid40661327, year = {2025}, author = {Kawakami, Y and Iguchi, Y and Li, J and Amakusa, Y and Yoshimura, T and Chikuchi, R and Yokoi, S and Iida, M and Riku, Y and Iwasaki, Y and Hirose, T and Nakagawa, S and Katsuno, M}, title = {Downregulation of NEAT1 due to loss of TDP-43 function exacerbates motor neuron degeneration in amyotrophic lateral sclerosis.}, journal = {Brain communications}, volume = {7}, number = {4}, pages = {fcaf261}, pmid = {40661327}, issn = {2632-1297}, abstract = {TAR DNA-binding protein 43 (TDP-43) is of particular interest in the pathogenesis of amyotrophic lateral sclerosis (ALS). It has been speculated that loss of nuclear TDP-43 and its cytoplasmic aggregation contributes to neurodegeneration. Although considerable attention has been paid to RNA metabolism in TDP-43 function, TDP-43 is also known to act as a transcription factor. This study found that the expression of Nuclear-enriched abundant transcript 1 (NEAT1), a long-non-coding RNA, was substantially downregulated in motor neurons with nuclear TDP-43 loss, but upregulated in those with preserved nuclear TDP-43, in the postmortem spinal cords of patients with sporadic ALS. TDP-43 depletion induced Neat1 downregulation in Neuro2a cells, primary cortical neurons, and mouse spinal motor neurons. Furthermore, TDP-43 was found to positively regulate NEAT1 at the transcriptional level. Finally, Neat1 knockout exacerbates neurodegeneration of hSOD1[G93A] mice accompanied by increased misfolded superoxide dismutase 1 (SOD1) aggregations. Transcriptome analysis revealed that Neat1 knockout reduced protein folding-related genes, such as heat shock protein family A member 1A (Hspa1a), in the spinal cords of hSOD1[G93A] mice. Our results indicated that the loss of TDP-43 function enhances ALS neurodegeneration by losing the protective effect of NEAT1.}, }
@article {pmid40683276, year = {2025}, author = {Harkness, JR and McDermott, JH and Marsden, S and Jamieson, P and Metcalfe, KA and Khan, N and Macken, WL and Pitceathly, RDS and Record, CJ and Maroofian, R and Kleopa, K and Christodoulou, K and Sabir, A and Islam, L and Santra, S and Durmusalioglu, EA and Atik, T and Isik, E and Cogulu, O and Urquhart, JE and Beaman, GM and Demain, LA and Jackson, A and Blakes, AJM and Byers, HJ and Bennett, H and Lin, WH and Adamson, A and Patel, S and Yue, WW and Taylor, RW and Reunert, J and Marquardt, T and Buchert, R and Haack, T and Losch, H and Ryba, L and Lassuthova, P and Valkovičová, R and Haberlová, J and Lauerová, B and Trúsiková, E and Polavarapu, K and Kilicarslan, OA and Lochmüller, H and Zamani, M and Chamanrou, N and Shariati, G and Sadeghian, S and Azizimalamiri, R and Maddirevula, S and AlMuhaizea, M and Alkuraya, FS and Horvath, R and Gungor, S and Manzur, A and Munot, P and Matthews, R and Banka, S and Reilly, MM and Bennett, D and O'Keefe, RT and Newman, WG}, title = {Acute-onset axonal neuropathy following infection in children with biallelic RCC1 variants: a case series.}, journal = {The Lancet. Neurology}, volume = {24}, number = {8}, pages = {667-680}, doi = {10.1016/S1474-4422(25)00198-X}, pmid = {40683276}, issn = {1474-4465}, mesh = {Adolescent ; Animals ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; *Cell Cycle Proteins/genetics ; *Infections/complications ; *Peripheral Nervous System Diseases/etiology ; Drosophila ; Nuclear Proteins ; Guanine Nucleotide Exchange Factors ; }, abstract = {BACKGROUND: The reasons why some individuals have severe neuropathy following an infection are not known. Through the agnostic screening of children with acute axonal neuropathy after an infection, we identified several families with biallelic variants in RCC1. We aimed to describe the clinical phenotype of these patients, and the molecular and cellular pathology associated with the genetic variants identified in these families.<br><br>METHODS: For this case series, we identified children affected by a severe, acute-onset axonal neuropathy following infection through an international research consortium of paediatric neurologists and clinical geneticists from nine countries (Canada, Cyprus, Czechia, Germany, Iran, Saudi Arabia, Slovakia, T&#xfc;rkiye, and the UK). Clinical assessments included nerve conduction studies and neuroimaging. We did exome or genome sequencing in DNA samples from all patients. We characterised the proteins encoded by the genetic variants by use of thermal stability and enzymatic assays, using recombinantly expressed proteins. We assessed cellular protein transport under heat or oxidative stress by use of immunofluorescence in primary fibroblasts, obtained from patients. We generated a humanised Drosophila knock-in model to assess the effects of stress on the in vivo function of RCC1.<br><br>FINDINGS: Between Nov 2, 2011, and July 10, 2024, we identified 24 individuals from 12 families who had severe, acute-onset axonal neuropathy following infection (13 female and 11 male patients, with a mean age at diagnosis of 1 year 10 months [SD 2&#xb7;27]). Eight biallelic missense variants in RCC1 were identified in affected individuals with autosomal recessive inheritance. Patients had variable phenotypes, ranging from rapidly progressive fatal axonal neuropathy to mild motor neuropathy with impaired walking. Neurological presentation was often secondary to an infection, resulting in initial misdiagnoses of Guillain-Barr&#xe9; syndrome in several patients. 15 children had disease recurrence. The disease was fatal in 15 patients. The RCC1 variants in these patients code for proteins that alter GDP-to-GTP exchange activity and have reduced thermal stability in vitro. In primary fibroblasts, heat shock or oxidative stress revealed defects in Ran nuclear localisation and impaired nucleocytoplasmic transport. A Drosophila model of the disease revealed a fatal intolerance to oxidative stress.<br><br>INTERPRETATION: We describe an autosomal recessive, acute-onset paediatric axonal neuropathy, seemingly triggered by infection, that affects individuals with biallelic RCC1 variants. In these children, the disease can mimic Guillain-Barr&#xe9; syndrome. The pathological mechanisms underlying this novel axonal neuropathy might overlap with those of amyotrophic lateral sclerosis. Cellular studies indicate that RCC1 variants affect nucleocytoplasmic transport, which is crucial for healthy axonal function. Future studies should be directed at pre-symptomatic treatment by exploring ways to maintain nucleocytoplasmic transport.<br><br>FUNDING: National Institute for Health and Care Research, LifeArc, and Wellcome Trust.}, }
@article {pmid40802071, year = {2025}, author = {Yamashita, T and Yokota, O and Ousaka, D and Sun, H and Haraguchi, T and Ota-Elliott, RS and Matsuoka, C and Kawano, T and Nakashima-Yasuda, H and Fukui, Y and Nakano, Y and Morihara, R and Hasegawa, M and Hosono, Y and Terada, S and Takaki, M and Ishiura, H}, title = {Biallelic variants in DNAJC7 cause familial amyotrophic lateral sclerosis with the TDP-43 pathology.}, journal = {Acta neuropathologica}, volume = {150}, number = {1}, pages = {19}, pmid = {40802071}, issn = {1432-0533}, support = {JP23K08543//Japan Society for the Promotion of Science/ ; JP23K10450//Japan Society for the Promotion of Science/ ; JP23K27514//Japan Society for the Promotion of Science/ ; JP24wm0425019//Japan Agency for Medical Research and Development/ ; JP23ek0109673//Japan Agency for Medical Research and Development/ ; }, mesh = {Adult ; Aged ; Animals ; Female ; Humans ; Male ; Middle Aged ; *Amyotrophic Lateral Sclerosis/genetics/pathology/metabolism ; Brain/pathology/metabolism ; *DNA-Binding Proteins/metabolism/genetics ; *HSP40 Heat-Shock Proteins/genetics ; Motor Neurons/pathology/metabolism ; Pedigree ; Zebrafish ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by the progressive degeneration of motor neurons. ALS pathology primarily involves the failure of protein quality control mechanisms, leading to the accumulation of misfolded proteins, particularly TAR DNA-binding protein 43 (TDP-43). TDP-43 aggregation is a central pathological feature of ALS. Maintaining protein homeostasis is critical and facilitated by heat shock proteins (HSPs), particularly the HSP40 family, which includes co-chaperones such as DNAJC7. Here, we report a family with three siblings affected by ALS who carry a homozygous c.518dupC frameshift variant in DNAJC7, a member of the HSP40 family. All three patients exhibited progressive muscle weakness, limb atrophy, bulbar palsy, and respiratory failure. Pathological examination revealed degeneration of both upper and lower motor neurons, with phosphorylated TDP-43-positive neuronal cytoplasmic inclusions in the frontal and temporal cortices. Immunoblot analysis were consistent with a type B pattern of phosphorylated TDP-43 in the precentral gyrus. Immunohistochemistry and RNA sequencing analyses demonstrated a substantial reduction in DNAJC7 expression at both the protein and RNA levels in affected brain regions. In a TDP-43 cell model, DNAJC7 knockdown impaired the disassembly of TDP-43 following arsenite-induced stress, whereas DNAJC7 overexpression suppressed the assembly and promoted the disassembly of arsenite-induced TDP-43 condensates. Furthermore, in a zebrafish ALS model, dnajc7 knockdown resulted in increased TDP-43 aggregation in motor neurons and reduced survival. To the best of our knowledge, this study provides the first evidence linking biallelic loss-of-function variants in DNAJC7 to familial ALS with TDP-43 pathology.}, }
@article {pmid40848171, year = {2026}, author = {Xu, H and Shao, Y and Zhang, J and Ni, Y and Xu, G and Liu, C and Liang, Y and Le, W}, title = {HSF-1 Regulates Autophagy to Govern Motor Function and Facilitate Toxic Protein Clearance in a C. elegans Model of Amyotrophic Lateral Sclerosis.}, journal = {Neuroscience bulletin}, volume = {42}, number = {4}, pages = {744-764}, pmid = {40848171}, issn = {1995-8218}, mesh = {Animals ; *Amyotrophic Lateral Sclerosis/metabolism/pathology/genetics ; Caenorhabditis elegans ; *Caenorhabditis elegans Proteins/metabolism/genetics ; *Autophagy/physiology ; Disease Models, Animal ; *Transcription Factors/metabolism/genetics ; Motor Neurons/metabolism/pathology ; Animals, Genetically Modified ; Signal Transduction ; Locomotion/physiology ; }, abstract = {Heat shock factor-1 (HSF-1) plays a crucial role in orchestrating stress responses across diverse organisms and disease conditions. Here, we investigate how the HSF-1 signaling pathway influences the degradation of toxic proteins and neuropathological changes in the Caenorhabditis elegans model of amyotrophic lateral sclerosis (ALS). We found that overexpressing HSF-1 improves locomotor ability and increases the survival rate of ALS C. elegans. Moreover, we observed a deceleration of motor neuron degeneration, demonstrating the protective effect of HSF-1 on neurodegenerative processes. Transcriptomic analysis revealed notable changes in genes associated with autophagy and neurodegeneration, underscoring HSF-1's critical involvement in ALS pathology. In addition, metabolomic profiling further highlighted the involvement of this pathway in metabolic reprogramming. Overall, our study underscores the critical role of the HSF-1 signaling pathway in improving survival rate, movement velocity, cellular integrity, and metabolic adaptation, providing new insights into the mechanisms underlying ALS and potential targets for therapeutic intervention.}, }
@article {pmid41005474, year = {2025}, author = {Li, Y and Liu, D and Li, S}, title = {IRE1/Xbp1 promotes the clearance of poly(GR) dipeptide repeats in amyotrophic lateral sclerosis.}, journal = {The Journal of biological chemistry}, volume = {301}, number = {11}, pages = {110764}, pmid = {41005474}, issn = {1083-351X}, mesh = {*Amyotrophic Lateral Sclerosis/metabolism/genetics/pathology ; Animals ; *Protein Serine-Threonine Kinases/metabolism/genetics ; Humans ; *X-Box Binding Protein 1/metabolism/genetics ; C9orf72 Protein/genetics/metabolism ; *Dipeptides/metabolism/genetics ; *Endoribonucleases/metabolism/genetics ; Mice ; Disease Models, Animal ; Signal Transduction ; Drosophila melanogaster ; }, abstract = {Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are neurodegenerative disorders characterized by the expansion of GGGGCC (G4C2) repeats in the C9orf72 gene and progressive motor neuron degeneration. A key pathological hallmark of these diseases is the accumulation and cytoplasmic mislocalization of dipeptide repeat (DPR) proteins, particularly poly(GR), which are neurotoxic. Enhancing the clearance of poly(GR) represents a promising therapeutic strategy; however, the molecular mechanisms regulating poly(GR) turnover are not fully understood. Our previous work demonstrated that translationally stalled poly(GR) is targeted by the ribosome-associated quality control (RQC) pathway. In the present study, we identify the IRE1/Xbp1s signaling axis as an essential regulator of poly(GR) degradation. Ectopic expression of IRE1 or its downstream effector Xbp1s, as well as pharmacological activation of IRE1 using IXA4, significantly reduces poly(GR) protein levels in a Drosophila disease model, mammalian cell lines, fibroblasts derived from patients with C9orf72-ALS, and a C9orf72 transgenic mouse model. Mechanistically, RNA-sequencing analysis reveals that IRE1/Xbp1s signaling upregulates heat shock protein Hsp70Ba, which plays a critical role in maintaining poly(GR) proteostasis. Additionally, we show that the Rictor/AKT/VCP pathway contributes to the translational regulation and turnover of poly(GR). Importantly, activation of IRE1, either through ectopic expression or IXA4 treatment, mitigates motor neuron loss in the C9orf72 mouse model. Collectively, our findings highlight the IRE1/Xbp1s axis as a key modulator of poly(GR) clearance and suggest its therapeutic potential in ALS/FTD.}, }
@article {pmid41007432, year = {2025}, author = {Ramos-Velasco, B and Alcalde, J and Izquierdo, JM}, title = {Welander Distal Myopathy-Associated TIA1 E384K Mutation Disrupts Stress Granule Dynamics Under Distinct Stress Conditions.}, journal = {Biology}, volume = {14}, number = {9}, pages = {}, pmid = {41007432}, issn = {2079-7737}, support = {RTI2018-098517B-I00 and PID2021-126152OB-I00//Ministerio de Ciencia, Innovaci&#xf3;n y Universidades and Agencia Espa&#xf1;ola de Investigaci&#xf3;n through FEDER funds (RTI2018-098517B-I00 and PID2021-126152OB-I00)/ ; }, abstract = {Cellular stress triggers the formation of diverse RNA-protein aggregates, which can be associated with physiological responses, pathological conditions, or even detrimental outcomes. Under stress-induced proteostasis disruption, these RNA-protein assemblies are known as stress granules (SGs). Targeting such condensates-while sparing functional RNAs and proteins-remains a major therapeutic challenge in protein aggregation disorders such as myopathies and neuropathies. In this study, we investigated the cellular response to various stress conditions in the context of the TIA1 E384K mutation, a founder variant implicated in both Welander distal myopathy (WDM) and amyotrophic lateral sclerosis (ALS). Cells were exposed to different stressors, including proteotoxic, proteostatic, chemotoxic, and osmotic insults, and the behavior of TIA1-related SGs was analyzed. Our findings reveal a distinct yet conserved pattern in the dynamics of TIA1-dependent SG formation and clearance, influenced by the specific type of stressor and modulated by eIF2α Ser35 phosphorylation. These results indicate that the WDM-associated TIA1 mutation leads to aberrant SG dynamics across different stress conditions. Collectively, these observations support the idea that TIA1 E384K-associated SG dysregulation plays a role in WDM and ALS pathogenesis and underscores the importance of multiple stress contexts in disease progression.}, }
@article {pmid41069215, year = {2026}, author = {Revolinski, SR and Amaral, M and Savic, M and Burke, IC}, title = {Genome-wide scan reveals CYP450 metabolism and stress response regulation underlying sulfosulfuron resistance in Bromus tectorum.}, journal = {Pest management science}, volume = {82}, number = {2}, pages = {1500-1508}, doi = {10.1002/ps.70300}, pmid = {41069215}, issn = {1526-4998}, support = {//U.S. Department of Agriculture/ ; //Washington Grain Commission/ ; }, mesh = {*Herbicide Resistance/genetics ; *Cytochrome P-450 Enzyme System/metabolism/genetics ; Genome-Wide Association Study ; *Sulfonamides/pharmacology ; *Herbicides/pharmacology ; Polymorphism, Single Nucleotide ; *Bromus/genetics/drug effects/metabolism/physiology ; Stress, Physiological/genetics ; Plant Proteins/genetics/metabolism ; Pyrimidines ; }, abstract = {BACKGROUND: Cheatgrass (Bromus tectorum L.) is a problematic weed species in the wheat cropping systems in the rainfed crop production areas of the inland Pacific Northwest (PNW) and now is becoming resistant to multiple modes of action. To identify mechanisms of non-target site acetohydroxy-acid/acetolactase synthase (AHAS/ALS) inhibitor resistance, 123 B. tectorum accessions were treated with three sulfosulfuron treatments (3.5, 35 and 350 g ha[-1]). A genome-wide association study (GWAS) was performed using the results from the spray trials to unravel the mechanisms underlying sulfosulfuron resistance in B. tectorum.<br><br>RESULTS: A single nucleotide polymorphism (SNP) explained up to 48% of observed phenotypic variation of sulfosulfuron resistance in B. tectorum but was not located near the AHAS/ALS gene in the genome. Candidate genes included members of the cytochrome P450 (CYP450) 71 gene family, heat shock-related proteins and a regulator of a heat shock-related protein. Additionally, the analysis revealed hormonal regulators, and genes involved in abiotic stress response as candidate genes.<br><br>CONCLUSION: Non-target site resistance for sulfosulfuron is present in B. tectorum populations of the inland PNW. A heat shock-related protein 70 regulator and an auxin response factor gene was near the SNP that explained 48% of the variation in the GWAS, indicating auxin regulation and stress response pathways are involved in the resistance of B. tectorum populations to sulfosulfuron. The heat shock-factor protein 70 regulated by the heat shock-related protein 70 regulator was found on another significant SNP and the SNPs those two genes were on have a significant interaction. Additionally, the GWAS analysis indicated CYP450 genes are likely involved in resistance. &#xa9; 2025 Society of Chemical Industry.}, }
@article {pmid41135683, year = {2025}, author = {Yeo, KH and Kong, JH and Ng, QH and Yoon, MJ and Agatha, O and Chae, E and Lee, HO and Je, HS and Choe, YJ}, title = {J-domain proteins cooperate with Hsp70 to drive multiphase separation of RNA-binding-deficient TDP-43.}, journal = {The Journal of biological chemistry}, volume = {301}, number = {12}, pages = {110854}, pmid = {41135683}, issn = {1083-351X}, mesh = {*HSP70 Heat-Shock Proteins/metabolism/genetics ; *Saccharomyces cerevisiae Proteins/metabolism/genetics/chemistry ; *Saccharomyces cerevisiae/metabolism/genetics ; *DNA-Binding Proteins/metabolism/genetics/chemistry ; *HSP40 Heat-Shock Proteins/metabolism/genetics ; Protein Domains ; *RNA-Binding Proteins/metabolism/genetics ; Molecular Chaperones/metabolism/genetics ; }, abstract = {RNA-free TDP-43, resulting from mutations or post-translational modifications in its RNA-binding domain, forms multiphase condensates with Hsp70 chaperones enriched in the core. The presence of this structure in the nucleus is thought to be associated with disease states. However, the mechanisms underlying its formation remain poorly understood. In particular, it is unclear whether J-domain proteins (JDPs), critical co-chaperones of Hsp70, are incorporated into multiphase condensates, and if so, how they contribute to the phase separation process. Using yeast as a model organism with a relatively small JDP family, we identified Sis1, but not Ydj1, as an important factor in TDP-43 multiphase separation. RNA-binding-deficient TDP-43 initially forms uniform condensates enriched with the JDP Sis1 but not with Hsp70. Subsequent recruitment of Hsp70 transforms these uniform structures into multiphase condensates, with both Sis1 and Hsp70 enriched in the core. This transition requires a functional J-domain, which stimulates Hsp70 ATPase activity. These findings reveal a key role for JDPs in TDP-43 multiphase separation and highlight JDP specificity in this process.}, }
@article {pmid41153395, year = {2025}, author = {De La Cerna, JLO and Talubo, NDD and Villanueva, BHA and Tsai, PW and Tayo, LL}, title = {Conserved Blood Transcriptome Patterns Highlight microRNA and Hub Gene Drivers of Neurodegeneration.}, journal = {Genes}, volume = {16}, number = {10}, pages = {}, pmid = {41153395}, issn = {2073-4425}, mesh = {*MicroRNAs/genetics/blood ; Humans ; *Neurodegenerative Diseases/genetics/blood ; *Transcriptome/genetics ; Gene Regulatory Networks ; Gene Expression Profiling ; RNA, Messenger/genetics ; Gene Expression Regulation ; }, abstract = {Background/Objectives: Neurodegenerative diseases (NDs) such as Alzheimer's (AD), Parkinson's (PD), Huntington's (HD), and Amyotrophic Lateral Sclerosis (ALS) are clinically distinct but share overlapping molecular mechanisms. Methods: To identify conserved systemic signatures, we analyzed blood RNA-Seq datasets using Weighted Gene Co-Expression Network Analysis (WGCNA), differential expression, pathway enrichment, and miRNA-mRNA network mapping. Results: Two modules, the red and turquoise, showed strong preservation across diseases. The red module was enriched for cytoskeletal and metabolic regulation, while the turquoise module involved immune, stress-response, and proteostatic pathways. Discussion: Key hub genes, such as HMGCR, ACTR2, MYD88, PTEN, EP300, and regulatory miRNAs like miR-29, miR-132, and miR-146a, formed interconnected networks reflecting shared molecular vulnerabilities. The absence of classical heat shock proteins in preserved blood modules highlights tissue-specific expression differences between blood and neural systems. Several hub genes overlap with known pharmacological targets, suggesting potential in translational relevance. Conclusions: Together, these findings reveal conserved blood-based transcriptional modules that suggest parallel central neurodegenerative processes and may support future biomarker development and possible therapeutic exploration.}, }
@article {pmid41155507, year = {2025}, author = {Sonkodi, B and Nagy, ZF and Keller-Pintér, A and Klivényi, P and Molnár, MJ and Széll, M}, title = {Genetic Variants in SDC3, KCNA2, KCNK1, KCNK16, and Heat Shock Transcription Factor-1 Genes: An Exploratory Analysis Supporting the Piezo2 Channelopathy Hypothesis in Amyotrophic Lateral Sclerosis Onset.}, journal = {International journal of molecular sciences}, volume = {26}, number = {20}, pages = {}, pmid = {41155507}, issn = {1422-0067}, mesh = {Humans ; *Amyotrophic Lateral Sclerosis/genetics/pathology ; *Ion Channels/genetics ; *Kv1.2 Potassium Channel/genetics ; *Heat Shock Transcription Factors/genetics ; Female ; Male ; Middle Aged ; *Channelopathies/genetics ; Genetic Predisposition to Disease ; Adult ; Genetic Variation ; Aged ; }, abstract = {Amyotrophic lateral sclerosis (ALS) is a multisystem progressive neurodegenerative disease. A recent theory of ALS onsetting pathogenesis proposed that the initiating primary damage is an acquired irreversible intrafusal proprioceptive terminal PIEZO2 channelopathy with underlying genetic and environmental risk factors. This Piezo2 channelopathy may also disrupt the ultrafast proton-based oscillatory signaling to motor neurons through vesicular transporter 1 (VGLUT1) and to the hippocampus through VGLUT2. As a result, it may gradually degenerate motor neurons in which process Kv1.2 ion channels are gradually depleted. It also gradually depletes heat shock transcription factor-1 (HSF-1) in the hippocampus, hence negatively affecting adult hippocampal neurogenesis. Syndecans, especially syndecan-3 (SDC3) in the nervous system, may act as critical players in the maintenance of the crosstalk between Piezo ion channels. Hence, our goal was to reanalyze the potential pathogenic gene variants from the cohort of our previous ALS study with a special focus on the aforementioned genes. Reanalysis of data formerly acquired by whole-exome sequencing of 21 non-related adult ALS patients was carried out with a focus on 28 genes. Accordingly, we identified charge-altering variants of SDC3 in 13 patients out of 21 that may contribute to the impairment of the Piezo crosstalk, and the progressive loss of the proposed proton-based signaling to motor neurons and to the hippocampus. A variant of uncertain significance was identified in the KCNA2 gene that may facilitate the faster loss of Kv1.2 ion function on motor neurons when Piezo2 channelopathy prevails. Not to mention that one variant was identified in the potassium current rectifying ion channels encoding KCNK1 and KCNK16 genes that may also propel the ALS disease process and provide the autoimmune-like pathogenic background. Moreover, Piezo2 channelopathy likely promotes diminishing HSF1 function in the hippocampus in the presence of the identified HSF1 variant. The current findings may support the ALS onsetting acquired irreversible Piezo2 channelopathy-induced pathogenesis. However, the preliminary nature of these findings needs validation and further functional studies on cohorts with a larger sample size in the future.}, }
@article {pmid41188870, year = {2025}, author = {Keerie, AF and Martins, RR and Allen, CF and Bowden, K and Al Mashhadi, S and Marlow, T and Myszczynska, M and Thakur, N and Beal, SN and Shaw, A and Suresh, S and McKinnon, SN and Cooper-Knock, J and West, RJH and Bonsall, S and Daniel, A and Wells, T and Kumar, V and Ellis, BCS and Higgins, M and Dinkova-Kostova, AT and Shelkovnikova, TA and Kalfus, IN and Shan, N and Shaw, PJ and Ferraiuolo, L and Mead, RJ}, title = {M102 activates both NRF2 and HSF1 transcription factor pathways and is neuroprotective in cell and animal models of amyotrophic lateral sclerosis.}, journal = {Molecular neurodegeneration}, volume = {20}, number = {1}, pages = {118}, pmid = {41188870}, issn = {1750-1326}, mesh = {Animals ; *Amyotrophic Lateral Sclerosis/metabolism/drug therapy ; *NF-E2-Related Factor 2/metabolism ; Mice, Transgenic ; Disease Models, Animal ; Mice ; Motor Neurons/drug effects/metabolism ; *Neuroprotective Agents/pharmacology ; *Heat Shock Transcription Factors/metabolism ; Humans ; Signal Transduction/drug effects ; }, abstract = {M102 is a central nervous system (CNS) penetrant small molecule electrophile which activates in vivo the NF-E2 p45-related factor 2-antioxidant response element (NRF2-ARE) pathway, as well as transcription of heat-shock element (HSE) associated genes. In the TDP-43[Q331K] transgenic mouse model of ALS dosed subcutaneously at 5 mg/kg OD or 2.5 mg/kg BD with M102, significant improvements in compound muscle action potential (CMAP) amplitude of hind limb muscles and gait parameters were observed at 6 months of age, with associated target engagement. An oral dose response study of M102 in SOD1[G93A] transgenic mice showed a dose-dependent improvement in CMAP of hindlimb muscles which correlated with preservation of lumbar spinal motor neurons at the same time point. These data enabled prediction of human efficacious exposures and doses, which were well within the safety margin predicted from Good Laboratory Practice (GLP) toxicology studies. A parallel program of work in vitro showed that M102 rescued motor neuron survival in co-culture with patient-derived astrocytes from sporadic, C9orf72 and SOD1 ALS cases. Markers of oxidative stress, as well as indices of TDP-43 proteinopathy were also reduced by exposure to M102 in these in vitro models. This comprehensive package of preclinical efficacy data across two mouse models as well as patient-derived astrocyte toxicity assays, provides a strong rationale for clinical evaluation of M102 in ALS patients. Combined with the development of target engagement biomarkers and the completed preclinical toxicology package, a clear translational pathway to testing in ALS patients has been developed.}, }
@article {pmid41294911, year = {2025}, author = {Abreu, MM and Hosseine-Farid, M and Silverman, DG}, title = {Total Reversal of ALS Confirmed by EMG Normalization, Structural Reconstitution, and Neuromuscular-Molecular Restoration Achieved Through Computerized Brain-Guided Reengineering of the 1927 Nobel Prize Fever Therapy: A Case Report.}, journal = {Diseases (Basel, Switzerland)}, volume = {13}, number = {11}, pages = {}, pmid = {41294911}, issn = {2079-9721}, abstract = {BACKGROUND: Neurological disorders are the leading cause of disability, affecting over three billion people worldwide. Amyotrophic lateral sclerosis (ALS) is among the most feared and uniformly fatal neurodegenerative diseases, with no therapy capable of restoring lost function.<br><br>METHODS: We report the first application of therapeutic fever to ALS using Computerized Brain-Guided Intelligent Thermofebrile Therapy (CBIT[2]). This fully noninvasive treatment, delivered through an FDA-approved computerized platform, digitally reengineers the 1927 Nobel Prize-recognized malarial fever therapy into a modern treatment guided by the Brain-Eyelid Thermoregulatory Tunnel. CBIT[2] induces therapeutic fever through synchronized hypothalamic feedback, activating heat shock proteins, which are known to restore proteostasis and neuronal function.<br><br>CASE PRESENTATION: A 56-year-old woman was diagnosed with progressive ALS at the Mayo Clinic, with electromyography (EMG) demonstrating fibrillation and fasciculation indicative of denervation corroborated by neurological and MRI findings; the patient was informed that she had an expected survival of three to five years. A neurologist from Northwestern University confirmed the diagnosis and thus maintained the patient on FDA-approved ALS drugs (riluzole and edaravone). Her condition rapidly worsened despite pharmacological treatment, and she underwent CBIT[2], resulting in (i) electrophysiological reversal with complete disappearance of denervation; (ii) biomarker correction, including reductions in neurofilament and homocysteine, IL-10 normalization (previously linked to mortality), and robust HSP70 induction; (iii) restoration of gait, swallowing, respiration, speech, and cognition; (iv) reconstitution of tongue structure; and (v) return to complex motor tasks, including golf, pickleball, and swimming.<br><br>DISCUSSION: This case provides the first documented evidence that ALS can be reversed through digitally reengineered fever therapy aligned with thermoregulation, which induces heat shock response and upregulates heat shock proteins, resulting in the patient no longer meeting diagnostic criteria for ALS and discontinuation of ALS-specific medications. Beyond ALS, shared protein-misfolding pathology suggests that CBIT[2] may extend to Alzheimer's, Parkinson's, and related disorders. By modernizing this Nobel Prize-recognized therapeutic principle with computerized precision, CBIT[2] establishes a framework for large-scale clinical trials. A century after fever therapy restored lost brain function and so decisively reversed dementia paralytica such that it earned the 1927 Nobel Prize in Medicine, CBIT[2] now safely harnesses the therapeutic power of fever through noninvasive, intelligent, brain-guided thermal modulation. Amid a global brain health crisis, fever-based therapies may offer a path to preserve thought, memory, movement, and independence for the more than one-third of humanity currently affected by neurological disorders.}, }
@article {pmid41309196, year = {2025}, author = {Jamwal, RS and Sharma, B and Minerva,  and Gupta, A and Misri, S and Shankaryan, R and Shah, R and Kumar, R}, title = {Protein misfolding and its dual role in neurodegeneration and cancer progression.}, journal = {Advances in protein chemistry and structural biology}, volume = {148}, number = {}, pages = {355-377}, doi = {10.1016/bs.apcsb.2025.10.001}, pmid = {41309196}, issn = {1876-1631}, mesh = {Humans ; *Neoplasms/metabolism/pathology ; *Protein Folding ; *Neurodegenerative Diseases/metabolism/pathology ; Animals ; Disease Progression ; Unfolded Protein Response ; }, abstract = {Protein misfolding is a fundamental biological process with profound implications for human health and disease. Typically, proteins assume precise three-dimensional structures to perform their functions, a process safeguarded by the proteostasis network, which comprises molecular chaperones, the ubiquitin-proteasome system (UPS), and autophagy. However, genetic mutations, oxidative stress, and environmental insults can disrupt folding, leading to the accumulation of non-functional or toxic conformations. In neurodegenerative diseases such as Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), Amyotrophic lateral Sclerosis (ALS), chronic misfolding results in toxic protein aggregates like amyloid-β, tau, and α-synuclein. These disrupt synaptic function, induce oxidative and nitrosative stress, and trigger apoptosis, ultimately leading to progressive neuronal loss. Dysregulation of the unfolded protein response (UPR) and weakened proteostasis with aging exacerbate disease pathology. In contrast, cancer cells utilize protein misfolding to enhance their survival and progression. Misfolded oncoproteins, such as mutant p53, not only evade degradation but also acquire oncogenic properties. Tumor cells hijack the UPR and chaperone networks, upregulate heat shock proteins, and manipulate oxidative stress responses to withstand hypoxia, nutrient deprivation, and rapid proliferation. Cancer stem cells (CSCs) further adapt to proteotoxic stress, contributing to tumor heterogeneity, therapy resistance, and immune evasion. The dual role of protein misfolding, driving degeneration in neurons while supporting proliferation in tumors, underscores its centrality in disease biology. Future research should focus on identifying early biomarkers of proteostasis imbalance and exploiting shared molecular pathways for the development of novel therapeutic interventions.}, }
@article {pmid41463293, year = {2025}, author = {Xie, V and Franco, MC and Martin, LJ}, title = {Human Mutant Dynactin Subunit 1 Causes Profound Motor Neuron Disease Consistent with Possible Mechanisms Involving Axonopathy, Mitochondriopathy, Protein Nitration, and T-Cell-Mediated Cytolysis.}, journal = {Biomolecules}, volume = {15}, number = {12}, pages = {}, pmid = {41463293}, issn = {2218-273X}, support = {R01 NS102479/NS/NINDS NIH HHS/United States ; 1R01NS107417-04A1/NH/NIH HHS/United States ; }, mesh = {Animals ; Humans ; Mice ; Anterior Horn Cells/metabolism/pathology ; *Axons/pathology ; *Dynactin Complex/genetics ; Mice, Transgenic ; *Mitochondrial Diseases/genetics ; *Motor Neuron Disease/genetics ; *Motor Neurons/metabolism/pathology ; Mutation ; *Protein Processing, Post-Translational/genetics ; Quinazolinones/pharmacology ; Spinal Cord Diseases ; T-Lymphocytes/immunology ; *Tyrosine/metabolism/analogs &amp; derivatives ; Mitochondrial Dynamics/drug effects ; Male ; Female ; }, abstract = {Mutations in the gene encoding the p150 subunit of the dynactin complex (DCTN1) are linked to amyotrophic lateral sclerosis, spinal and bulbar muscular atrophy, and Perry syndrome. These neurodegenerative diseases can cause muscle weakness and atrophy, parkinsonian-like symptoms, and paralysis. To examine the evolution of neuropathology caused by a mutation in DCTN1 and cellular mechanisms of disease for therapeutic discovery, we characterized mice expressing either human wildtype or mutant (G59S) DCTN1. Neuron-specific expression of mutant, but not wildtype, DCTN1 caused fatal age-related paralytic disease and motor neuron (MN) degeneration in the spinal cord with axonopathy and chromatolysis without apoptotic morphology. MNs became positive for cleaved caspase-3, cleaved caspase-8, and nitrated Hsp90. Mitochondria accumulated and appeared fragmented and dysmorphic and then were lost. This pathology was accompanied by invasion of CD95- and CD8-positive mononuclear T cells into the ventral horn and accumulation of TNFα and IL9. Administration of the mitochondrial division inhibitor-1 (Mdivi-1) protected MNs and extended the lifespan of G59S-DCTN1 mice. A mitochondrial permeability transition pore inhibitor also extended lifespan. Thus, mutant DCTN1 causes degeneration of MNs associated with axonopathy, mitochondriopathy, nitrative stress, and caspase activation. It appears as retrograde neurodegeneration and inflammatory T-cell-like cytolysis. Mitochondria are possible therapeutic targets in DCTN1-linked neurodegenerative disorders.}, }
@article {pmid41572777, year = {2026}, author = {Liu, X and Lv, Z and Xu, G and Chen, Y and Liu, H and Xu, P}, title = {Investigating the Multiple Regulatory Mechanisms and Therapeutic Targets of PHLDA1 in Neurological Diseases.}, journal = {Current neuropharmacology}, volume = {}, number = {}, pages = {}, doi = {10.2174/011570159X413561251125064110}, pmid = {41572777}, issn = {1875-6190}, abstract = {PHLDA1 (pleckstrin homology-like domain family A member 1) is a pleiotropic regulatory protein that affects key biological processes such as apoptosis, pyroptosis, immune inflammation, autophagy, metabolism, and oxidative stress. PHLDA1 plays a significant role in the pathological mechanisms of neurological diseases. This article systematically reviews the molecular characteristics of PHLDA1 and its core role in cerebrovascular diseases such as cerebral ischemia/ reperfusion injury, cerebral hemorrhage, subarachnoid hemorrhage, epilepsy, amyotrophic lateral sclerosis (ALS), and Parkinson's disease (PD). Studies have shown that PHLDA1 promotes disease progression by regulating signalling pathways such as the NF-κB, MAPK, NLRP3 inflammasome, PPARγ, and Nrf2 pathways, thereby exacerbating neuroinflammation, mitochondrial dysfunction, endoplasmic reticulum stress, and pyroptosis in neurons. Its expression is regulated by the dynamic balance of miRNAs (such as miR-194 and miR-101), transcription factors (Egr1 and BHLHE40), and heat shock proteins (HSPs/HSF1). In addition, PHLDA1 has become a potential target for intervention in neurodegenerative and ischemic injuries by inhibiting FundC1-mediated mitochondrial autophagy, regulating microglial polarization, and activating TRAF6-dependent neuroinflammation. This article not only clarifies the pathogenic mechanism of PHLDA1 but also summarizes the relevant intervention strategies targeting PHLDA1, hoping to provide a corresponding theoretical basis and reference for the development of precision therapies for neurological diseases.}, }
@article {pmid41767843, year = {2026}, author = {Ben Khalaf, N}, title = {Heat shock proteins (Hsp70 and Hsp90) in neurodegeneration: pathogenic roles and therapeutic potential.}, journal = {Frontiers in aging neuroscience}, volume = {18}, number = {}, pages = {1711422}, pmid = {41767843}, issn = {1663-4365}, abstract = {The maintenance of protein homeostasis is essential for neuronal survival and function; however, it progressively declines with age, predisposing the brain to neurodegenerative diseases. Molecular chaperones Hsp70 and Hsp90 are key guardians of proteostasis, pivotally regulating protein folding, refolding, and degradation under both physiological and stress conditions. This review integrates an overview of the structural features, isoforms, and mechanistic interactions of Hsp70 and Hsp90. It highlights how their dysfunction contributes to the pathogenesis of major neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and Huntington's disease. We first examine the architecture and ATP-driven chaperone cycles of Hsp70 and Hsp90, their co-chaperone networks, and the feedback regulation by the Heat Shock Factor-1 pathway. We then discuss evidence linking age-related declines in chaperone expression and HSF-1 activity to proteostasis collapse and neuronal vulnerability. The review particularly examines how Hsp70 and Hsp90 differentially influence pathogenic protein aggregation (e.g., tau, α-synuclein, TDP-43, and mutant huntingtin) and how this balance is altered in the aging brain. Regarding therapeutic approaches, we summarize current strategies targeting these chaperones, including small-molecule modulators of Hsp70 and Hsp90, co-chaperone inhibitors, and recombinant chaperone therapy, which has shown to restore proteostasis and cognitive function in experimental models. These emerging interventions underscore the dual nature of Hsp70/Hsp90 systems, acting as both protectors and potential contributors to neurodegeneration, depending on their regulation and interaction context. By linking molecular chaperone biology to aging and translational therapeutics, this review establishes a framework for developing precision approaches that enhance proteostasis capacity, delay age-associated neurodegeneration, and promote healthy brain aging.}, }
@article {pmid41854301, year = {2026}, author = {Walker, TB and Trowbridge, JW and McMahon, S and Marzano, NR and Rice, L and Yerbury, JJ and Ecroyd, H and McAlary, L}, title = {Small heat shock proteins HspB1 and HspB5 differentially alter the condensation and aggregation of the TDP-43 low-complexity domain.}, journal = {Protein science : a publication of the Protein Society}, volume = {35}, number = {4}, pages = {e70539}, pmid = {41854301}, issn = {1469-896X}, support = {//Motor Neurone Disease Australia/ ; //FightMND/ ; APP1194872//National Health and Medical Research Council/ ; }, mesh = {Humans ; *DNA-Binding Proteins/chemistry/metabolism/genetics ; Protein Aggregates ; *HSP27 Heat-Shock Proteins/metabolism/chemistry/genetics ; *alpha-Crystallin B Chain/metabolism/chemistry/genetics ; Molecular Chaperones/metabolism ; Protein Domains ; Phosphorylation ; Heat-Shock Proteins ; }, abstract = {TAR DNA-binding protein 43 (TDP-43) is a nucleic acid-binding protein that regulates processes of mRNA metabolism, during which it undergoes condensation mediated by its C-terminal low-complexity domain (TDP-43[LCD]). TDP-43 aggregation and condensation are associated with neurodegenerative disease. However, the proteostasis mechanisms that regulate these processes remain elusive. Some evidence has shown that the molecular chaperone small heat shock protein HspB1 binds to and regulates the cytoplasmic phase separation of TDP-43, indicating that other small heat shock proteins may have similar effects. Here, we demonstrate divergent behaviors for HspB1 and its homolog HspB5 on TDP-43[LCD] condensation and aggregation. In addition to inhibiting TDP-43[LCD] aggregation, HspB1 partitions into TDP-43[LCD] condensates and increases the dynamic exchange of TDP-43[LCD] within condensates and with the surrounding solution. Phosphorylation-mimicking mutations within HspB1 enhance these effects. HspB5 inhibits TDP-43[LCD] aggregation more effectively than HspB1 and partitions into TDP-43[LCD] condensates, where it delays the pathological transition of the condensate to a gel/solid. We identify the N- and C-terminal regions of HspB1 and HspB5 to be crucial for the chaperone effects, and highlight the role of sequence diversity within these regions in defining small heat shock protein function. These findings demonstrate that HspB1 and HspB5 are regulators of TDP-43 phase separation and aggregation and may be potential therapeutic targets in mitigating toxic TDP-43 aggregation in neurodegenerative disease.}, }