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DOI: 10.1160/TH04-12-0783
Fibrillar type I collagens enhance platelet-dependent thrombin generation via glycoprotein VI with direct support of α2β1 but not αIIbβ3 integrin
Publication History
Received
07 December 2004
Accepted after resubmission
28 April 2004
Publication Date:
05 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200501/lookinside/thumbnails/10-1160-th04-12-0783-1.jpg)
Summary
The role of collagens and collagen receptors was investigated in stimulating platelet-dependent thrombin generation. Fibrillar type-I collagens, including collagen from human heart, were most potent in enhancing thrombin generation, in a way dependent on exposure of phosphatidylserine (PS) at the platelet surface. Soluble, non-fibrillar type-I collagen required pre-activation of integrin α2β1 with Mn2+ for enhancement of thrombin generation. With all preparations, blocking of glycoprotein VI (GPVI) with 9O12 antibody abrogated the collagen-enhanced thrombin generation, regardless of the α2β1 activation state. Blockade of α2β1 alone or antagonism of autocrine thromboxane A2 and ADP were less effective. Blockade of αIIbβ3 with abciximab suppressed thrombin generation in platelet-rich plasma, but this did not abolish the enhancing effect of collagens. The high activity of type-I fibrillar collagens in stimulating GPVI-dependent procoagulant activity was confirmed in whole-blood flow studies, showing that these collagens induced relatively high expression of PS. Together, these results indicate that: i) fibrillar type-I collagen greatly enhances thrombin generation, ii) GPVI-induced platelet activation is principally responsible for the procoagulant activity of fibrillar and non-fibrillar collagens, iii) α2β1 and signaling via autocrine mediators facilitate and amplify this GPVI activity, and iv) αIIbβ3 is not directly involved in the collagen effect.
* Authors contributed equally to this work
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References
- 1 Ruggeri ZM. Platelets in atherothrombosis. Nat Med 2002; 8: 1227-34.
- 2 Nieswandt B, Watson SP. Platelet-collagen interaction: is GPVI the central receptor?. Blood 2003; 102: 449-61.
- 3 Nieswandt B, Brakebusch C, Bergmeier W. et al. Glycoprotein VI but not α2β1 integrin is essential for platelet interaction with collagen. EMBO J 2001; 20: 2120-30.
- 4 Atkinson BT, Jarvis GE, Watson SP. Activation of GPVI by collagen is regulated by α2β1 and secondary mediators. J Thromb Haemost 2003; 1: 1278-87.
- 5 Chen H, Kahn ML. Reciprocal signaling by integrin and nonintegrin receptors during collagen activation of platelets. Mol Cell Biol 2003; 23: 4764-77.
- 6 Siljander PR, Munnix ICA, Smethurst PA. et al. Platelet receptor interplay regulates collagen-induced thrombus formation in flowing human blood. Blood 2004; 103: 1333-41.
- 7 Savage B, Almus-Jacobs F, Ruggeri ZM. Specific synergy of multiple substrate-receptor interactions in platelet thrombus formation under flow. Cell 1998; 94: 657-66.
- 8 Savage B, Ginsberg MH, Ruggeri ZM. Influence of fibrillar collagen structure on the mechanisms of platelet thrombus formation under flow. Blood 1999; 94: 2704-15.
- 9 Jung SM, Moroi M. Platelet collagen receptor integrin α2β1 activation involves differential participation of ADP-receptor subtypes P2Y1 and P2Y12 but not intracellular calcium change. Eur J Biochem 2001; 268: 3513-22.
- 10 Lecut C, Schoolmeester A, Kuijpers MJ. et al. Principal role of glycoprotein VI in α2β1 and αIIbβ3 activation during collagen-induced thrombus formation. Arterioscler Thromb Vasc Biol 2004; 24: 1727-33.
- 11 Heemskerk JWM, Bevers EM, Lindhout T. Platelet activation and blood coagulation. Thromb Haemost 2002; 88: 186-93.
- 12 Monroe DM, Hoffman M, Roberts HR. Platelets and thrombin generation. Arterioscler Thromb Vasc Biol 2002; 22: 1381-9.
- 13 Heemskerk JWM, Siljander P, Vuist WMJ. et al. Function of glycoprotein VI and integrin α2β1 in the procoagulant response of single, collagen-adherent platelets. Thromb Haemost 1999; 81: 782-92.
- 14 Alberio L, Safa O, Clemetson KJ. et al. Surface expression and functional characterization of α-granule factor V in human platelets. Blood 2000; 95: 1694-702.
- 15 Furihata K, Clemetson KJ, Deguchi H. et al. Variation in human platelet glycoprotein VI content modulates glycoprotein VI-specific prothrombinase activity. Arterioscler Thromb Vasc Biol 2001; 21: 1857-63.
- 16 Bevers EM, Comfurius P, van Rijn JLML. et al. Generation of prothrombin-converting activity and the exposure of phosphatidylserine at the outer surface of platelets. Eur J Biochem 1982; 122: 429-36.
- 17 Andersen H, Greenberg DL, Fujikawa K. et al. Protease- activated receptor 1 is the primary mediator of thrombin-stimulated platelet procoagulant activity. Proc Natl Acad Sci USA 1999; 96: 11189-93.
- 18 Siljander P, Farndale RW, Feijge MAH. et al. Platelet adhesion enhances the glycoprotein VI-dependent procoagulant response. Arterioscler Thromb Vasc Biol 2001; 21: 618-27.
- 19 Farndale RW, Sixma JJ, Barnes MJ. et al. The role of collagen in thrombosis and hemostasis. J Thromb Haemost 2004; 2: 561-73.
- 20 Siljander P, Lassila R. Studies of adhesion-dependent platelet activation. Arterioscler Thromb Vasc Biol 1999; 19: 3033-43.
- 21 Lecut C, Feeney LA, Kingsbury G. et al. Human platelet glycoprotein VI function is antagonized by monoclonal antibody-derived Fab fragments. J Thromb Haemost 2003; 1: 2653-62.
- 22 Miller EJ, Rhodes RK. Preparation and characterization of the different types of collagen. Meth Enzymol 1982; 82: 33-64.
- 23 Hemker HC, Giesen PLA, Ramjee M. et al. The thrombogram: monitoring thrombin generation in platelet-rich plasma. Thromb Haemost 2000; 83: 589-91.
- 24 Vanschoonbeek K, Feijge MAH, van Kampen RJ. et al. Initiating and potentiating role of platelets in tissue factor-induced thrombin generation in the presence of plasma. J Thromb Haemost 2004; 2: 476-84.
- 25 Kuijpers MJE, Schulte V, Bergmeier W. et al. Complementary roles of glycoprotein VI and α2β1 integrin in collagen-induced thrombus formation in flowing whole blood ex vivo . FASEB J 2003; 17: 685-7.
- 26 Andree HA, Reutelingsperger CP, Hauptmann R. et al. Binding of vascular anticoagulant α (VACα) to planar phospholipid bilayers. J Biol Chem 1990; 265: 4923-8.
- 27 Léon C, Ravanat C, Freund M. et al. Differential involvement of the P2Y1 and P2Y12 receptor in platelet procoagulant activity. Arterioscler Thromb Vasc Biol 2003; 23: 1941-7.
- 28 Dorsam RT, Tuluc M, Kunapuli SP. Role of protease- activated and ADP receptor subtypes in thrombin generation on human platelets. J Thromb Haemost 2004; 2: 804-12.
- 29 Bazzoni G, Shih DT, Buck CA. et al. Monoclonal antibody 9EG7 defines a novel β1 integrin epitope induced by soluble ligand and manganese, but inhibited by calcium. J Biol Chem 1995; 270: 25570-77.
- 30 Humphries MJ. Integrin structure. Biochem Soc Trans 2000; 28: 311-39.
- 31 Jung SM, Moroi M. Platelets interact with soluble and insoluble collagens through characteristically different reactions. J Biol Chem 1998; 273: 14827-37.
- 32 Jandrot-Perrus M, Lagrue AH, Okuma M. et al. Adhesion and activation of human platelets induced by convulxin involve glycoprotein VI and integrin α2β1. J Biol Chem 1997; 272: 27035-41.
- 33 Reverter JC, Béguin S, Kessels H. et al. Inhibition of platelet-mediated, tissue-factor-induced thrombin generation by the mouse/human chimeric 7E3 antibody. J Clin Invest 1996; 98: 863-74.
- 34 Béguin S, Kumar R, Keularts I. et al. Fibrin-dependent platelet procoagulant activity requires GPIb receptors and von Willebrand factor. Blood 1999; 93: 564-70.
- 35 Butenas S, Cawthern KM, van 't Veer C. et al. Antiplatelet agents in tissue factor-induced blood coagulation. Blood 2001; 97: 2314-22.
- 36 Ilveskero S, Lassila R. Abciximab inhibits procoagulant activity but not the release reaction upon collagen- or clot-adherent platelets. J Thromb Haemost 2003; 1: 805-13.
- 37 Weber KT, Sun Y, Tyagi SC. et al. Collagen network of the myocardium. J Mol Cell Cardiol 1994; 26: 279-92.
- 38 Cosemans JMEM, Kuijpers MJE, Lecut C. et al. Contribution of platelet glycoprotein VI to the thrombogenic effect of collagens in fibrous atherosclerotic lesions. Atherosclerosis, in press. doi: 10.1016/j.atherosclerosis. 2004.12.037.
- 39 Kunapuli SP, Dorsam RT, Kim S. et al. Platelet purinergic receptors. Curr Opin Pharmacol 2003; 3: 175-80.
- 40 Auger JM, Kuijpers MJE, Senis YA. et al. Adhesion of human and mouse platelets to collagen under shear: a unifying model. FASEB J 2005; 19: 825-7.