Intracellular matrix metalloproteinase-2 (MMP-2) regulates human platelet activation via hydrolysis of talin

Summary

Matrix metalloproteinase (MMP) activity is generally associated with normal or pathological extracellular processes such as tissue remodeling in growth and development or in tumor metastasis and angiogenesis. Platelets contain at least three MMPs, 1, 2 and 9 that have been reported to stimulate or inhibit agonist-induced platelet aggregation via extracellular signals. The non-selective Zn⁺² chelating MMP inhibitor, 1,10-phenanthroline, and the serine protease inhibitor, AEBSF, were found to inhibit all tested agonist-induced platelet aggregation reactions. In vitro analysis demonstrated that 1,10-phenanthroline completely inhibited MMP-1,2,and 9 but had little to no effect on calpain activity while the converse was true with AEBSF. We now demonstrate that MMP-2 functions intracellularly to regulate agonistinduced platelet aggregations via the hydrolytic activation of talin, the presumed final activating factor of glycoprotein (GP)IIb/IIIa integrin (the inside-out signal). Once activated GPIIb/IIIa binds the dimeric fibrinogen molecule required for platelet aggregation. The active intracellular MMP-2 molecule is complexed with JAK 2/STAT 3, as demonstrated by the fact that all three proteins are co-immunoprecipitated with either anti-JAK 2, or anti-STAT 3 antibodies and by immunofluorescence studies. The MMP-2 platelet activation pathway can be synergistically inhibited with the non-selective MMP inhibitor, 1,10-phenanthroline, plus a JAK 2 inhibitor. This activation pathway is distinct from the previously reported calpain-talin activating pathway. The identification of a new central pathway for platelet aggregation presents new potential targets for drug regulation and furthers our understanding of the complexity of platelet activation mechanisms.

Keywords

Matrix metalloproteinase - platelet physiology - thrombin - talin

• References

• 1 Hadler-Olsen E, Fadnes B, Sylte I. et al. Regulation of matrix metalloproteinases activity in health and disease. FEBS J 2011; 278: 28-45.
• 2 Gialeli C, Theocharis AD. Karamanos NK. Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J 2011; 278: 16-27.
• 3 Back M, Ketelhuth DF, Agewall S.. Matrix Metalloproteinases in atherothrombosis. Progress in Cardiovascular Diseases 2010; 52: 410-428.
• 4 Cunningham LA, Wezel M, Rosenberg GA. Multiple roles for MMPs and TIMPs in cerebral ischemia. Glia 2005; 50: 329-339.
• 5 Candelario-Jalil E, Yang Y, Rosenberg GA. Diverse roles of matrix metalloproteinases and tissue inhibitors or metalloproteinases in neuroinflammation and cerebral ischemia. Neuroscience 2009; 158: 983-994.
• 6 Sawicki G, Salas E, Murat J. et al. Release of gelatinase A during platelet activation mediates aggregation. Nature 1997; 386: 616-619.
• 7 Fernandez-Patron C, Martinez-Cuesta MA, Salas E. et al. Differential regualtion of platelet aggregation by matrix metalloproteinases-9 and -2.. Thromb Haemost 1999; 82: 1730-1735.
• 8 Kazes I, Elalamy I, Sraer J-D. et al. Platelet release of trimolecular complex components MT1-MMP/TIMP2/MMP2 involvement in MMP2 activation and platelet aggregation. Blood 2000; 96: 3064-3069.
• 9 Galt SW, Lindemann S, Allen L. et al. Outside-In signals delivered by matrix metalloproteinase-1 to regulate platelet function. Circ Res 2002; 90: 1093-1099.
• 10 Falcinelli E, Gugliemini G, Torti M. et al. Intraplatelet signalling mechanisms of the matrix metalloproteinase-2 on platelet aggregation. J Thromb Haemost 2005; 03: 2526-2535.
• 11 Lee YM, Lee JJ, Shen MY. et al. Inhibitory mechanisms of activated matrix metalloproteinases-9 on platelet activation. Eur Pharmacol 2006; 537: 52-58.
• 12 Choi W S, Jeon O H, Kim H. et al. MMP-2 regulates human platelet activation by interacting with integrin (IIb(3. J Thromb Haemost 2008; 06: 517-523.
• 13 Santos-Martinez MJ, Medina C, Jurasz P. et al. Role of metalloproteinases in platelet function Thromb Res 2008; 121: 535-542.
• 14 Santos-Martinez MJ, Medina C, Glimer JF. et al. Matrix metalloproteinases in platelet function: coming of age. J Thromb Haemost 2008; 06: 514-516.
• 15 Juarasz P, Chung AWY, Radomski A. et al. Nonremodelling properties properties of matrix metalloproteinases. The platelet connection. Circ Res 2002; 90: 1041-1043.
• 16 Sheu JR, Fong TH, Liu CM. et al. Expression of matrix metalloproteinase-9 in human platelets: regulation of platelet activation in in vitro and in vivo studies. Pharmacol 2004; 143: 191-201.
• 17 Galt SW, Lindemann S, Allen L. et al. Outside-in signals delivered by matrix metalloproteinase-1 regulate platelet function. Circ Res 2002; 90: 1093-1099.
• 18 Falcinelli E, Bury L, Tolley N. et al. MMP-9 in platelets: maybe, maybe not. Blood 2011; 118: 6471-6473.
• 19 Kalvegren H, Jonsson S, Jonasson L.. Release of matrix metalloproteinases-1 and-2, but not -9, from activated platelets measured by enzyme-linked immunosorbent assay. Platelets 2011; 22: 572-578.
• 20 Trivedi V, Boire A, Tchernychev B. et al. Platelet matrix metalloproteinase-1 mediates thrombogenesis by activating PAR1 at a cryptic ligand site. Cell 2009; 137: 332-343.
• 21 Shattil SJ, Kashiwagi H, Pampori N. Integrin signalling: the platelet paradigm. Blood 1998; 91: 2645-2657.
• 22 Petrich BG, Marchese P, Ruggeri ZM. et al. Talin is required for integrin-mediated platelet function in hemostasis and thrombosis. JEM 2007; 204: 3103-3111.
• 23 Azam M, Andrabi SS, Sahr KE. et al. Disruption of the mouse μ-calpain gene reveals an essential role in platelet function. Molec Cell Biol 2001; 21: 2213-2220
• 24 Hayashi M, Suzuki H, Kawashima S. et al. The behavior of calpain-generated Nand C-terminal fragments of talin in integrin-mediated signalling pathways. Arch Biochem Biophys 1999; 371: 133-141.
• 25 Schoenwaelder SM, Yuan Y, Cooray P. et al. Calpain cleavage of focal adhesion proteins regulates the cytoskeletal attachment of integrin alphaIIbbeta 3 (platelet glycoprotein IIb/IIIa) and the cellular retraction of fibrin clots. J Biol Chem 1997; 272: 1694-1702.
• 26 Yan B, Calderwood DA, Yaspan B. et al. Calpain cleavage promotes talin binding to the β3 integrin cytoplasmic domain. J Biol Chem 2001; 276: 2864-2870.
• 27 Schulz R. Intracellular targets of matrix metalloproteinase-2 in cardiac disease: rationale and therapeutic approaches. Ann Rev Pharmacol Toxicol 2007; 47: 211-242.
• 28 Moshal KS, Tipparaju SM, Vacek TP. et al. Mitochandrial matrix metalloproteinase activation decreases myocyte contractility in hyperhomocysteinemia. Am J Physiol Heart Circ Physiol 2008; 295: H890-H897
• 29 McCawley LJ, Matrisian LM. Matrix metalloproteinases: they’re not just for matrix anymore!. Curr Opinion in Cell Biol 2001; 13: 534-540.
• 30 Shapiro D, Senior RM. Matrix Metalloproteinase. Matrix degradation and more. AM J Respir Cell Molec Biol 1999; 20: 1100-1102.
• 31 Duncan ME, Richardson JP, Murray GI. et al. Human matrix metalloproteinase-9 Activation by limited trypsin treatment and generation of monoclonal antibodies specific for the activated form. Eur J Biochem 1998; 258: 37-43.
• 32 Grant GA, Eisen AZ, Marmer BL. et al. The activation of human skin fibroblast procollagenase. Sequence identification of the major conversion products. J Biol Chem 1987; 262: 5886-5889.
• 33 Galis ZS, Kranzhofer R, Fenton II JW. et al. Thrombin promotes activation of matrix metalloproteinase-2 produced by cultured vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1997; 17: 483-489.
• 34 Tompa P, Buzder-Lantos P, Tantos A. et al. On the sequential determinants of calpain, cleavage. J Biol Chem 2004; 279: 20775-20785.
• 35 Rosenfeldt MT, Valentino M, Labruzzo S. et al. The organomecurial 4-aminophenylmercuric acetate, independent of matrix metalloproteinases induces dosedependent activation/inhibition of platelet aggregation. Thromb Haemost 2005; 93: 326-330.
• 36 Soslau G, Class R, Morgan DA. et al. Unique pathway of thrombin-induced platelet aggregation mediated by glycoprotein Ib. J Biol Chem 2001; 276: 21173-21183.
• 37 Soslau G, Pastorino MB, Morgan DA. et al. Herpes simplex virus replication and protein synthesis in a human blood-derived cell line. J Gen Virol 1987; 68: 2079-2092.
• 38 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685.
• 39 Soslau G, Wallace B, Vicente C. et al. Comparison of functional aspects of the coagulation cascade in human and sea turtle plasmas. Comp Biochem Physiol, Part B 2004; 138: 399-406.
• 40 Soslau G, Giles J. The loss of sialic acid and its prevention in stored human platelets. Thrombosis Res 1982; 26: 443-455.
• 41 Soslau G, Goldenberg SJ, Class R. et al. Differential activation and inhibition of human platelet thrombin receptors by structurally distinct α-, β- and γ-thrombin. Platelets 2004; 15: 155-166.
• 42 Nieswandt B, Moser M, Pleines I. et al. Loss of talin 1 in platelets abrogates integrin activation platelet aggregation, and thrombus formation in vitro and in vivo. JEM 2007; 204: 3113-3118.
• 43 Ezumi Y, Takayama H, Okuma M. Thrombopoietin, c-Mpl ligand, induces tyrosine phosphorylation of TyK2, JAK2, and STAT3, and enhances agonist-induced aggregation in vitro. FEBS Lett 1995; 374: 48-52.
• 44 Chen K, Rondina MT, Weyrich AS. A sticky story for signal transducer and activator of transcription 3 in platelets. Circulation 2013; 127: 421-423.
• 45 Brodsky I, Siegel NH, Kahn SB. et al. Simultaneous fibrinogen and platelet survival with [75 Se] selenomethionine in man. Br J Haematol 1970; 18: 341-355.
• 46 Soslau G, Rybicki A. In vitro incorporation of fucose and methionine into human platelet proteins. Biochem Biophys Res Commun 1982; 109: 1256-1263.
• 47 Soslau G, Morgan DA, Jaffe JS. et al. Cytokine mRNA expression in human platelets and a megakaryocyte cell line and cytokine modulation of platelet function. Cytokine 1997; 9: 405-411.
• 48 Brass LF, Manning DR, Cichowski K. et al. Signalling through G proteins in platelets: to the integrins and beyond. Thromb Haemost 1997; 78: 5810589.
• 49 Yang J, Wu J, Jiang H. et al. Signalling through Gi family members in platelets. Redundancy and specificity in the regulation of adenylyl cyclase and other effectors. J Biol Chem 2002; 277: 46035-46042.
• 50 Zhang L, Brass LF, Manning DR. The Gq and G12 families of heterotrimeric G proteins report functional selectivity. Mol Pharmacol 2009; 75: 235-241.
• 51 Malinin NL, Plow EF. Byzova TV Kindlins in FERM adhesion. Blood 2010; 115: 4011-4017.
• 52 Cecchetti L, Tolley ND, Mitchetti N. et al. Megakaryocytes differentially sort mRNAs for matrix metalloproteinases and their inhibitors into platelets: a mechanism for regulating synthetic events. Blood 2011; 118: 1903-1911.
• 53 Wrzyszcz A, Wozniak M. On the origin of matrix metalloproteinase-2 and -9 in blood platelets. Platelets 2012; 23: 467-474.
• 54 Weyrich AS, Schwertz H, Kraiss LW, Zimmerman GA. Protein synthesis by platelets: Historical and new perspectives. J Thromb Haemost 2009; 7: 241-246.
• 55 Schulze H, Ballmaier M, Welte K. et al. Thrombopoietin induces the generation of distinct Stat 1, Stat 3, Stat 5a and Stat 5b homo- and heterodimeric complexes with different kinetics in human platelets. Exp Hematol 2000; 28: 294-304.
• 56 Momi S, Falcinelli E, Giannini S. et al. Loss of matrix metalloproteinase-2 in platelets reduces arterial thrombosis in vivo. JEM 2009; 206: 2365-2379.