Abstract
Understanding the neuropathology of multiple sclerosis (MS) is essential for improved therapies. Therefore, identification of targets specific to pathological types of MS may have therapeutic benefits. Here we identify, by laser-capture microdissection and proteomics, proteins unique to three major types of MS lesions: acute plaque, chronic active plaque and chronic plaque. Comparative proteomic profiles identified tissue factor and protein C inhibitor within chronic active plaque samples, suggesting dysregulation of molecules associated with coagulation. In vivo administration of hirudin or recombinant activated protein C reduced disease severity in experimental autoimmune encephalomyelitis and suppressed Th1 and Th17 cytokines in astrocytes and immune cells. Administration of mutant forms of recombinant activated protein C showed that both its anticoagulant and its signalling functions were essential for optimal amelioration of experimental autoimmune encephalomyelitis. A proteomic approach illuminated potential therapeutic targets selective for specific pathological stages of MS and implicated participation of the coagulation cascade.
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References
Lassmann, H., Bruck, W. & Lucchinetti, C. Heterogeneity of multiple sclerosis pathogenesis: implications for diagnosis and therapy. Trends Mol. Med. 7, 115–121 (2001)
Frohman, E. M., Racke, M. K. & Raine, C. S. Multiple sclerosis – the plaque and its pathogenesis. N. Engl. J. Med. 354, 942–955 (2006)
Wekerle, H. Immune pathogenesis of multiple sclerosis. Neurol. Sci. 26 (suppl. 1). S1–S2 (2005)
Benito, C. et al. Cannabinoid CB1 and CB2 receptors and fatty acid amide hydrolase are specific markers of plaque cell subtypes in human multiple sclerosis. J. Neurosci. 27, 2396–2402 (2007)
Kagitani-Shimono, K. et al. Lipocalin-type prostaglandin D synthase (beta-trace) is upregulated in the αB-crystallin-positive oligodendrocytes and astrocytes in the chronic multiple sclerosis. Neuropathol. Appl. Neurobiol. 32, 64–73 (2006)
Minn, A. J. et al. Genes that mediate breast cancer metastasis to lung. Nature 436, 518–524 (2005)
Rubinsztein, D. C. & Serra, H. G. Protein–protein interaction networks in the spinocerebellar ataxias. Genome Biol. 7, 229.1–229.3 (2006)
Sam-Yellowe, T. Y. et al. A Plasmodium gene family encoding Maurer’s cleft membrane proteins: structural properties and expression profiling. Genome Research 14, 1052–1059 (2004)
Chabas, D. et al. The influence of the proinflammatory cytokine, osteopontin, on autoimmune demyelinating disease. Science 294, 1731–1735 (2001)
John, G. R. et al. Multiple sclerosis: re-expression of a developmental pathway that restricts oligodendrocyte maturation. Nature Med. 8, 1115–1121 (2002)
Lock, C. et al. Gene-microarray analysis of multiple sclerosis lesions yields new targets validated in autoimmune encephalomyelitis. Nature Med. 8, 500–508 (2002)
Peng, J., Elias, J. E., Thoreen, C. C., Licklider, L. J. & Gygi, S. P. Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome. J. Proteome Res. 2, 43–50 (2003)
Hwang, S. I. et al. Systematic characterization of nuclear proteome during apoptosis: a quantitative proteomic study by differential extraction and stable isotope labeling. Mol. Cell. Proteomics 5, 1131–1145 (2006)
Lundgren, D. H., Eng, J., Wright, M. E. & Han, D. K. PROTEOME-3D: an interactive bioinformatics tool for large-scale data exploration and knowledge discovery. Mol. Cell. Proteomics 2, 1164–1176 (2003)
Esmon, C. T. Crosstalk between inflammation and thrombosis. Maturitas 47, 305–314 (2004)
Eddleston, M. et al. Astrocytes are the primary source of tissue factor in the murine central nervous system. A role for astrocytes in cerebral hemostasis. J. Clin. Invest. 92, 349–358 (1993)
Riewald, M. & Ruf, W. Orchestration of coagulation protease signaling by tissue factor. Trends Cardiovasc. Med. 12, 149–154 (2002)
Mosnier, L. O., Zlokovic, B. V. & Griffin, J. H. The cytoprotective protein C pathway. Blood 109, 3161–3172 (2007)
Song, X., Huhle, G., Wang, L., Hoffmann, U. & Harenberg, J. Generation of anti-hirudin antibodies in heparin-induced thrombocytopenic patients treated with r-hirudin. Circulation 100, 1528–1532 (1999)
de Kleijin, E. D. et al. Activation of protein C following infusion of protein C concentrate in children with severe meningococcal sepsis and purpura fulminans: a randomized, double-blinded, placebo-controlled, dose-finding study. Crit. Care Med. 31, 1839–1847 (2003)
Laterre, P. F. et al. Severe community-acquired pneumonia as a cause of severe sepsis: data from the PROWESS study. Crit. Care Med. 33, 952–961 (2005)
Bernard, G. R. et al. Efficacy and safety of recombinant human activated protein C for severe sepsis. N. Engl. J. Med. 344, 699–709 (2001)
Kirichuk, V. F. & Streknev, A. G. Rol’ sistemy gemostaza v patogeneze i techenii rasseiannogo skleroza. Zh. Nevrol. Psikhiatr. Im. S S Korsakova (special issue 2). 103, 34–38 (2003)
Gerlitz, B. G. Mutation of protease domain residues Lys37–39 in human protein C inhibits activation by the thrombomodulin-thrombin complex without affecting activation by free thrombin. J. Biol. Chem. 271, 22285–22288 (1996)
Grinnell, B. W. et al. Differentiation of cytoprotective vs. anticoagulant function with variants of activated protein C in LPS-induced renal microvascular dysfunction. Crit. Care Med. 35, (suppl.) abstr. 42 (2007)
Preston, R. J. et al. Multifunctional specificity of the protein C/activated protein C Gla domain. J. Biol. Chem. 281, 28850–28857 (2006)
Cheng, T. et al. Activated protein C inhibits tissue plasminogen activator-induced brain haemorrhage. Nature Med. 12, 1278–1285 (2006)
Steinman, L. Blocking adhesion molecules as therapy for multiple sclerosis: natalizumab. Nature Rev. Drug Discov. 4, 510–518 (2005)
Maschmeyer, J., Shearer, R., Lonser, E. & Spindle, D. Heparin potassium in the treatment of chronic multiple sclerosis. Bull. Los Angel. Neuro. Soc. 26, 165–171 (1961)
Lider, O. et al. Suppression of experimental autoimmune diseases and prolongation of allograft survival by treatment of animals with low doses of heparins. J. Clin. Invest. 83, 752–756 (1989)
Bagnato, C. et al. Proteomics analysis of human coronary athrosclerosis plaque: a feasibility study of direct tissue proteomics by liquid chromatography and tandem mass spectrometry. Mol. Cell. Proteomics 6, 1088–1102 (2007)
Yates, J. R., Eng, J. K., McCormack, A. L. & Schieltz, D. Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal. Chem. 67, 1426–1436 (1995)
Han, D. K., Eng, J., Zhou, H. & Aebersold, R. Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry. Nature Biotechnol. 19, 946–951 (2001)
Liu, H., Sadygov, R. G. & Yates, J. R. A model for random sampling and estimation of relative protein abundance in shotgun proteomics. Anal. Chem. 76, 4193–4201 (2004)
Peng, J. et al. A proteomics approach to understanding protein ubiquitination. Nature Biotechnol. 21, 921–926 (2003)
Platten, M. et al. Treatment of autoimmune neuroinflammation with a synthetic tryptophan metabolite. Science 310, 850–855 (2005); erratum 311, 954 (2006)
Greenfield, E. A. et al. Monoclonal antibodies to distinct regions of human myelin proteolipid protein simultaneously recognize central nervous system myelin and neurons of many vertebrate species. J. Neurosci. Res. 83, 415–431 (2006)
Ousman, S. S. et al. Protective and therapeutic role for αB-crystallin in autoimmune demyelination. Nature 448, 474–479 (2007)
Acknowledgements
We thank Mary Jane Eaton for help with histopathology and Jian Luo for help with imaging microscopy. This work was funded by the National Institutes of Health and the National Multiple Sclerosis Society to L.S., the National Institutes of Health to D.H., and Ruth L. Kirschstein National Research Service Award and T32 Adult and Pediatric Rheumatology and Immunology Fellowship awards to M.H.H. B.W.G. and B.G. are employed by Lilly Research Laboratories, a division of Eli Lilly & Co.
Author Contributions M.H.H. and L.S. formulated the hypothesis and designed all the experiments. D.K.H., S.-I.H. and D.H.L. contributed the proteomic studies. D.B.R. performed the EAE experiment with hirudin treatment. R.A.S. and C.S.R. contributed to the histopathological analysis. B.G. and B.W.G. provided the recombinant aPC proteins. J.V.P. performed studies on NF-κB signalling. S.S.O. performed the in vitro assays with astrocytes. D.K.H. and L.S., the senior authors, contributed equally to this work.
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Brian W. Grinnell and Bruce Gerlitz are employees of Eli Lilly.
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Han, M., Hwang, SI., Roy, D. et al. Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets. Nature 451, 1076–1081 (2008). https://doi.org/10.1038/nature06559
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DOI: https://doi.org/10.1038/nature06559
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