Blockade of Platelet GPIIB-IIIA (Integrin αIIbβ3) in Flowing Human Blood Leads to Passivation of Prothrombotic Surfaces

Markus A. Riederer; Mark H. Ginsberg; Beat Steiner

doi:10.1055/s-0037-1613314

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 88(05): 858-864
DOI: 10.1055/s-0037-1613314

Review Article

Schattauer GmbH

Blockade of Platelet GPIIB-IIIA (Integrin α_IIbβ₃) in Flowing Human Blood Leads to Passivation of Prothrombotic Surfaces

Markus A. Riederer

²Department of Vascular and Metabolic Diseases, F. Hoffmann-La Roche Ltd., Basel, Switzerland

,

Mark H. Ginsberg

¹Departments of Vascular Biology and Cell Biology, The Scripps Research Institute, La Jolla, California, USA

,

Beat Steiner

²Department of Vascular and Metabolic Diseases, F. Hoffmann-La Roche Ltd., Basel, Switzerland

› Author Affiliations

Abstract

Summary

We examined the impact of platelet activation on platelet adhesion to collagen in flowing human blood. ADP activation of platelets in ex vivo flowing blood resulted in paradoxical inhibition of platelet deposition on collagen. Blockade of fibrinogen binding to platelets by Lamifiban, a competitive antagonist of GPIIb-IIIa (integrin α_IIbβ₃), reversed this inhibition, leading to a marked increase in integrin α₂β₁-dependent platelet adhesion. Analysis of integrin α₂β₁-dependent platelet adhesion to collagen indicated that ADP-induced suppression of platelet adhesion is the result of trans-dominant inhibition of integrin α₂β₁ caused by fibrinogen binding to integrin GPIIb-IIIa. Lamifiban blocked fibrinogen binding, reversing the trans-dominant inhibition of α₂β₁ dependent adhesion to collagen. The GPIIb-IIIa antagonist resulted in the formation of a nonthrombogenic, passivated surface comprised of an adherent platelet monolayer. This unexpected consequence of blocking fibrinogen binding to GPIIb-IIIa may explain the long-term benefits of short-term GPIIb-IIIa antagonist treatment of Acute Coronary Syndrome patients.

Keywords

Platelet - flow - GPIIb-IIIa - α₂β₁ - trans-dominant inhibition

Full Text

References

References
1 Sixma JJ, Van Zanten GH, Saelmann EUM, Verkleij M, Lankhof H, Nieuwenhus HK, De Groot PG. Platelet adhesion. Thromb Haemost 1995; 74: 454.
2 Shattil SJ, Kashiwagi H, Pampori N. Integrin signaling: the platelet paradigm. Blood 1998; 91: 2645.
3 Gresham HD, Goodwin JL, Anderson DC, Brown EJ. A novel member of the integrin receptor family mediates Arg-Gly-Asp-stimulated neutrophil phagocytosis. J Cell Biol 1989; 108: 1935.
4 Chen YP, O’Toole TE, Shipley T, Forsyth J, LaFlamme SE, Yamada KM, Shattil SJ, Ginsberg MH. “Inside-out” signal transduction inhibited by isolated integrin cytoplasmic domains. J Biol Chem 1994; 269: 18307.
5 Pacifici R, Roman J, Kimble R, Civitelli R, Brownfield CM, Bizzarri C. Ligand binding to monocyte alpha5 beta 1 integrin activates the alpha 2 beta 1 receptor via the alpha 5 subunit cytoplasmic domain and protein kinase C. J Immunol 1994; 153: 2222.
6 Díaz-González F, Forsyth J, Steiner B, Ginsberg MH. Trans-dominant inhibition of integrin function. Mol Biol Cell 1996; 07: 1939.
7 Poter JC, Hogg N. Integrin cross talk: activation of lymphocyte functionassociated antigen-1 on human T cells alters alpha4 beta1and alpha5 beta1-mediated function. J Cell Biol 1997; 138: 1437.
8 Fitzgerald DJ, Roy L, Catella F, Fitzgerald GA. Platelet activation in unstable coronary disease. N Engl J Med 1986; 315: 983.
9 Theroux P, Latour JG, Leger-Gauthier C, DeLara J. Fibrinopeptide A and platelet factor levels in unstable angina pectoris. Circulation 1987; 75: 156.
10 Ault KA, Cannon CP, Mitchell J, McCahan J, Tracy RP, Novotny WF, Reimann JD, Braunwald E. Platelet activation in patients after an acute coronary syndrome: results from the TIMI-12 trial. Thrombolysis in myocardial infarction. J Am Coll Cardiol 1999; 33: 634.
11 Becker RC, Bovill EG, Corrao JM, Ball SP, Ault K, Mann K, Tracy RP. Platelet activation determined by flow cytometry persists despite antithrombotic therapy in patients with unstable angina and Non-Q-wave myocardial infarction. J Thromb Thrombolysis 1994; 01: 95.
12 Paragon, and Investigators. International, randomized, controlled trial of lamifiban (a platelet glycoprotein IIb/IIIa antagonism for the reduction of acute coronary syndrome events in a gobal organization net work. Circulation 1998; 97: 2386.
13 Zhao XQ, Theroux P, Snapinn SM, Sax FL. Intracoronary thrombus and platelet glycoprotein IIb/IIIa receptor blockade with tirofiban in unstable angina or non-Q-wave myocardial infarction. Angiographic results from the PRISM-PLUS trial (Platelet receptor inhibition for ischemic syndrome management in patients limited by unstable signs and symptoms). Circulation 1999; 100: 16009.
14 Mahaffey KH, Harrington RA, Simoons ML, Granger CB, Graffagnino C, Alberts MJ, Laskowitz DT, Miller JM, Sloan MA, Berdan LG, MacAulay CM, Lincoff AM, Deckers J, Topol EJ, Califf RM. Stroke in patients with acute coronary syndromes: incidence and outcomes in the platelet glycoprotein IIb/IIIa in unstable angina. Receptor suppression using integrilin therapy (PURSUIT) trial. Circulation 1999; 99: 2371.
15 Kouns WC, Hadvary P, Haering P, Steiner B. Conformational modulation of purified glycoprotein (GP) IIb-IIIa allows proteolytic generation of active fragments from either active or inactive GP IIb-IIIa. J Biol Chem 1992; 267: 18844.
16 Steiner B, Haering P, Jennings L, Kouns WC. Five independent neoepitopes on GPIIb-IIIa are differentially exposed by two potent peptidomimetic platelet inhibitors. Thromb Haemost 1993; 69: 782.
17 Goding JW. Fragmentation of monoclonal antibodies. Monoclonal antibodies: Principles and Practice. Academic Press; 2nd edition. 1986: 125.
18 Alig L, Edenhofer A, Hadváry P, Hürzeler M, Knopp D, Müller M, Steiner B, Trzeciak A, Weller T. Low molecular weight, non-peptide fibrinogen receptor antagonists. J Med Chem 1992; 35: 4393.
19 Hilpert K, Ackermann J, Banner DW, Gast A, Gubernator K, Hadváry P, Labler L, Müller K, Schmid G, Tschopp TB, Van de Waterbeemd H. Design and synthesis of potent and highly selective thrombin inhibitors. J Med Chem 1994; 37: 3889.
20 Kirchhofer D, Tschopp TB, Steiner B, Baumgartner HR. Role of collagenadherent platelets in mediating fibrin formation in flowing whole blood. Blood 1995; 86: 3815.
21 Santoro SA, Zutter MM. The α₂β₁ integrin: a collagen receptor on platelets and other cells. Thromb Haemost 1995; 74: 813.
22 Saelman EUM, Nieuwenhius HK, Hese KM, De Groot PG, Jeijnene HFG, Sage EH, Wiliams S, McKeown L, Gralnick HR, Sixma JJ. Platelet adhesion to collagen types I through VIII under conditions of stasis and flow is mediated by GPIa/IIa (α₂β₁-integrin). Blood 1994; 83: 1244.
23 Holme S, Sixma JJ, Wester J, Holmsen H. ADP-induced refractory state of platelets in vitro. II. Functional and ultra studies of gel filtered platelets. Scand J Haematol 1977; 18: 267.
24 Frelinger ALI, Lam SC, Plow EF, Smith MA, Loftus JC, Ginsberg MH. Occupancy of an adhesive glycoprotein receptor modulates expression of an antigenic site involved in cell adhesion. J Biol Chem 1988; 263: 12397.
25 Blystone SD, Graham IL, Lindberg FP, Brown EJ. Integrin α_vβ₃ differentially regulates adhesive and phagocytic functions of the fibronectin receptor α₅β₁ . J Cell Biol 1994; 127: 1129.
26 Nakamura T, Kambayashi J, Okuma M, Tandon NN. Activation of the GPIIb-IIIa complex induced by platelet adhesion to collagen is mediated by both α₂β₁ integrin and GP VI. J Biol Chem 1999; 274: 11897.
27 Remijn JA, Wu YP, Ijsseldijk JW, Zwaginga JJ, Sixma JJ, de Groot PG. Absence of Fibrinogen in Afibrinogenemia Results in Large but Loosely Packed Thrombi under Flow Conditions. Thromb Haemost 2001; 85: 736.
28 Sakariassen KS, Nievelstein PFEM, Coller BS, Sixma JJ. The role of platelet membrane glycoproteins Ib and IIb-IIa in platelet adherence to human artery subendothelium. Br J Haematol 1996; 63: 681-91.
29 Roald HE, Sakariassen KS. Axial dependence of collagen-induced thrombus formation in flowing non-anticoagulated human blood. Anti-platelet drugs impair thrombus growth and increase platelet-collagen adhesion. Thromb Haemost 1995; 73: 126.