Cystatin C colocalizes with amyloid‐β and coimmunoprecipitates with amyloid‐β precursor protein in sporadic inclusion‐body myositis muscles

Cystatin C (CC), an endogenous cysteine protease inhibitor, is accumulated within amyloid‐β (Aβ) amyloid deposits in Alzheimer's disease (AD) brain and was proposed to play a role in the AD pathogenesis. Because the chemo‐morphologic muscle phenotype of sporadic inclusion‐body myositis (s‐IBM) has several similarities with the phenotype of AD brain, including abnormal accumulation of Aβ deposits, we studied expression and localization of CC in muscle biopsies of 10 s‐IBM, and 16 disease‐ and five normal‐control muscle biopsies. Physical interaction of CC with amyloid‐β precursor protein (AβPP) was studied by a combined immunoprecipitation/immunoblotting technique in the s‐IBM muscle biopsies and in AβPP‐overexpressing cultured human muscle fibers. In all s‐IBM muscle biopsies, CC‐immunoreactivity either colocalized with, or was adjacent to, the Aβ‐immunoreactive inclusions in 80–90% of the vacuolated muscle fibers, mostly in non‐vacuolated regions of their cytoplasm. Ultrastructurally, CC immunoreactivity‐colocalized with Aβ on 6–10 nm amyloid‐like fibrils and floccular material. By immunoblotting, CC expression was strongly increased in IBM muscle as compared to the controls. By immunoprecipitation/immunoblotting experiments, CC coimmunoprecipitated with AβPP, both in s‐IBM muscle and in AβPP‐overexpressing cultured normal human muscle fibers. Our studies (i) demonstrate for the first time that CC physically associates with AβPP, and (ii) suggest that CC may play a novel role in the s‐IBM pathogenesis, possibly by influencing AβPP processing and Aβ deposition.

[1]  T. Morgan,et al.  Vaccination with soluble Aβ oligomers generates toxicity‐neutralizing antibodies , 2001, Journal of neurochemistry.

[2]  J. Growdon,et al.  Elevation of cystatin C in susceptible neurons in Alzheimer's disease. , 2001, The American journal of pathology.

[3]  D. Selkoe.  Alzheimer's disease: genes, proteins, and therapy. , 2001, Physiological reviews.

[4]  W. Engel,et al.  Difference in Expression of Phosphorylated Tau Epitopes between Sporadic Inclusion‐body Myositis and Hereditary Inclusion‐body Myopathies , 1996, Journal of neuropathology and experimental neurology.

[5]  W. Engel,et al.  Expression of muscle-specific isozymes of phosphorylase and creatine kinase in human muscle fibers cultured aneurally in serum-free, hormonally/chemically enriched medium , 1990, International Journal of Developmental Neuroscience.

[6]  Philippe Taupin,et al.  FGF-2-Responsive Neural Stem Cell Proliferation Requires CCg, a Novel Autocrine/Paracrine Cofactor , 2000, Neuron.

[7]  Z. Simmons,et al.  Sporadic inclusion body myositis and hereditary inclusion body myopathy. , 2002, Journal of clinical neuromuscular disease.

[8]  B. Ghetti,et al.  Codeposition of Cystatin C with Amyloid‐β Protein in the Brain of Alzheimer Disease Patients , 2001, Journal of neuropathology and experimental neurology.

[9]  D. Small,et al.  Regulation of APP cleavage by alpha-, beta- and gamma-secretases. , 2000, FEBS letters.

[10]  J. Buxbaum,et al.  Presence of BACE1 and BACE2 in muscle fibres of patients with sporadic inclusion-body myositis , 2001, The Lancet.

[11]  W. Engel,et al.  Use of anti‐neurofilament antibody to identify paired‐helical filaments in inclusion‐body myositis , 1996, Annals of neurology.

[12]  W. Engel,et al.  β‐Amyloid precursor epitopes in muscle fibers of inclusion body myositis , 1993 .

[13]  W. Engel,et al.  Strong immunoreactivity of β-amyloid precursor protein, including the β-amyloid protein sequence, at human neuromuscular junctions , 1992, Neuroscience Letters.

[14]  R. Nitsch,et al.  Increased Cystatin C in Astrocytes of Transgenic Mice Expressing the K670N-M671L Mutation of the Amyloid Precursor Protein and Deposition in Brain Amyloid Plaques , 2001, Neurobiology of Disease.

[15]  N. Robakis,et al.  Distinct Secretases, a Cysteine Protease and a Serine Protease, Generate the C Termini of Amyloid β‐Proteins Aβ1‐40 and Aβ1‐42, Respectively , 1999 .

[16]  I. Ekiel,et al.  Folding-related Dimerization of Human Cystatin C (*) , 1996, The Journal of Biological Chemistry.

[17]  C. Tovar,et al.  Control and expression of cystatin C by mouse decidual cultures , 2002, Molecular reproduction and development.

[18]  D. Small,et al.  Regulation of APP cleavage by α‐, β‐ and γ‐secretases , 2000 .

[19]  A. Grubb.  Cystatin C-Properties and use as diagnostic marker , 2001, Advances in Clinical Chemistry.

[20]  I. Mellman,et al.  Developmental Regulation of Invariant Chain Proteolysis Controls MHC Class II Trafficking in Mouse Dendritic Cells , 1998, Cell.

[21]  Zbigniew Grzonka,et al.  Human cystatin C, an amyloidogenic protein, dimerizes through three-dimensional domain swapping , 2001, Nature Structural Biology.

[22]  D. Selkoe,et al.  Evidence that the 42- and 40-amino acid forms of amyloid beta protein are generated from the beta-amyloid precursor protein by different protease activities. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[23]  W. Engel,et al.  βAPP gene transfer into cultured human muscle induces inclusion‐body myositis aspects , 1997 .

[24]  J. Ghiso,et al.  Amyloid fibrils in hereditary cerebral hemorrhage with amyloidosis of Icelandic type is a variant of gamma-trace basic protein (cystatin C). , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[25]  W. Engel,et al.  beta-Amyloid precursor epitopes in muscle fibers of inclusion body myositis. , 1993, Annals of neurology.

[26]  G. Lynch,et al.  Lysosomal Protease Inhibitors Induce Meganeurites and Tangle-like Structures in Entorhinohippocampal Regions Vulnerable to Alzheimer's Disease , 1999, Experimental Neurology.

[27]  W. Engel,et al.  Enhanced detection of Congo‐red‐positive amyloid deposits in muscle fibers of inclusion body myositis and brain of Alzheimer's disease using fluorescence technique , 1993, Neurology.

[28]  Pilar Latorre,et al.  Alzheimer’s disease and the cystatin C gene polymorphism: an association study , 2001, Neuroscience Letters.

[29]  W. Engel,et al.  Light and electron microscopic immunolocalization of presenilin 1 in abnormal muscle fibers of patients with sporadic inclusion-body myositis and autosomal-recessive inclusion-body myopathy. , 1998, The American journal of pathology.

[30]  Masashi Yamada,et al.  Involvement of cystatin C in oxidative stress-induced apoptosis of cultured rat CNS neurons , 2000, Brain Research.

[31]  W. Engel,et al.  beta-Amyloid precursor protein mRNA is increased in inclusion-body myositis muscle. , 1993, Neuroreport.

[32]  INTERNATIONAL SOCIETY FOR NEUROCHEMISTRY , 1976 .

[33]  N. Robakis,et al.  Distinct secretases, a cysteine protease and a serine protease, generate the C termini of amyloid beta-proteins Abeta1-40 and Abeta1-42, respectively. , 1999, Journal of neurochemistry.

[34]  R. Roos,et al.  Comparison between the Icelandic and Dutch forms of hereditary cerebral amyloid angiopathy , 1992, Clinical Neurology and Neurosurgery.

[35]  W. Engel,et al.  Transfer of beta-amyloid precursor protein gene using adenovirus vector causes mitochondrial abnormalities in cultured normal human muscle. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[36]  W. Engel,et al.  Inclusion-body myositis and myopathies: different etiologies, possibly similar pathogenic mechanisms , 2002, Current opinion in neurology.

[37]  B T Hyman,et al.  Genetic association of a cystatin C gene polymorphism with late-onset Alzheimer disease. , 2000, Archives of neurology.

[38]  M. J. Conroy,et al.  Three‐dimensional domain swapping in the folded and molten‐globule states of cystatins, an amyloid‐forming structural superfamily , 2001, The EMBO journal.

[39]  W. Engel,et al.  Novel Immunolocalization of α‐Synuclein in Human Muscle of Inclusion‐Body Myositis, Regenerating and Necrotic Muscle Fibers, and at Neuromuscular Junctions , 2000, Journal of neuropathology and experimental neurology.