Anti-thrombotic action of clopidogrel and PlA1/A2 polymorphism of β3 integrin in patients with coronary artery disease not being treated with aspirin

Jerzy Dropinski; Jacek Musial; Bogdan Jakiela; Wojciech Wegrzyn; Marek Sanak; Andrew Szczeklik

doi:10.1160/TH05-05-0359

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2005; 94(06): 1300-1305
DOI: 10.1160/TH05-05-0359

Cell Signalling and Vessel Remodelling

Schattauer GmbH

Anti-thrombotic action of clopidogrel and Pl^A1/A2 polymorphism of β₃ integrin in patients with coronary artery disease not being treated with aspirin

Jerzy Dropinski

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

,

Jacek Musial

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

,

Bogdan Jakiela

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

,

Wojciech Wegrzyn

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

,

Marek Sanak

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

,

Andrew Szczeklik

¹Department of Medicine, Jagiellonian University School of Medicine, Krakow, Poland

› Institutsangaben

Abstract

Summary

Individual variability in response to clopidogrel is known but its mechanism is poorly understood. We examined the relationship between glycoprotein IIIa polymorphism Pl^A1/A2 and anti-thrombotic actions of clopidogrel. Clopidogrel (75 mg/d; 2 weeks) was administered to 48 normolipemic patients with coronary artery disease. Bleeding time, thrombin generation at the site of microvascular injury, platelet function under high shear, using PFA-100 with ADP cartridge, and platelet surface activation markers (P-selectin and fibrinogen binding sites on GPIIb/IIIa complex detected by PAC-1 antibody), were studied both before and after clopidogrel treatment. Both unstimulated and low-dose (0.02 μM and 1 μM) in vitro ADP-stimulated platelets were examined. GP IIIa polymorphism was assessed by polymerase chain reaction and restriction fragment length polymorphism analysis. We identified 32 Pl^A1/A1 homozygotes, 15 Pl^A1/A2 heterozygotes and one Pl^A2/A2 homozygote. Clopidogrel significantly prolonged bleeding time in all subjects, but this effect was greater in Pl^A2 carriers (p<0.01). Furthermore, clopidogrel only depressed thrombin generation at the site of microvascular injury (p<0.01) in Pl^A2 patients and prolonged closure time measured in vitro by PFA-100 (p<0.05). At baseline spontaneous expression of PAC-1 and P-selectin was higher in Pl^A2 subjects as compared to Pl^A1 homozygotes (p<0.05 for both antigens). Clopidogrel lowered the expression of both markers affecting more Pl^A2 carriers, so that the difference in binding PAC-1 antibody between platelets from Pl^A1 and Pl^A2 carriers disappeared, while the difference in P-selectin expression slightly diminished. Anti-thrombotic effects of clopidogrel are more pronounced in CAD patients carrying the Pl^A2 allele than in Pl^A1 homozygotes.

Keywords

Clopidogrel - coronary artery disease - glycoprotein IIIa polymorphism - platelet activation - thrombin generation

Volltext

Referenzen

References
1 Fuster V, Badimon L, Badimon JJ. et al The pathogenesis of coronary artery disease and the acute coronary syndromes. N Engl J Med 1992; 326: 310-8.
2 Trip MD, Cats VM, van Capelle FJ. et al Platelet hyperactivity and prognosis in survivors of myocardial infarction. N Eng J Med 1990; 322: 1549-54.
3 Hirsh J, Colman RW, Marder VJ. et al Overview of thrombosis and its treatment. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN. Hemostasis and thrombosis: basic principles and clinical practise. Philadelphia: Lippincott Williams and Wilkins; 2001: 1071-84.
4 Calvete JJ. Clues for understanding the structure and function of a prototypic human integrin: the platelet glycoprotein IIb/IIIa complex. Thromb Haemost 1994; 72: 1-15.
5 Coller BS. Anti-GPIIb/IIIa drugs: current strategies and future directions. Thromb Haemost 2001; 86: 427-43.
6 Newman PJ, Derbes RS, Aster RH. The human platelet alloantigens, P1A1 and PlA2, are associated with a leucine33/proline33 amino acid polymorphism in membrane glycoprotein IIIa, and are distinguishable by DNA typing. J Clin Invest 1989; 83: 1778-81.
7 von dem Borne AE, Decary F. Nomenclature of platelet-specific antigens. Transfusion 1990; 30: 477.
8 Di Castelnuovo A, de Gaetano G, Donati MB. et al Platelet glycoprotein receptor IIIa polymorphism Pl^A1/Pl^A2 and coronary risk: a meta-analysis. Thromb Haemost 2001; 85: 626-33.
9 Zhu MM, Weedon J, Clark LT. Meta-analysis of the association of platelet glycoprotein IIIa PlA1/A2 polymorphism with myocardial infarction. Am J Cardiol 2000; 86: 1000-5.
10 Dorsam RT, Kim S, Jin J. et al Coordinated signaling through both G12/13 and Gi pathways is sufficient to activate GPIIb/IIIa in human platelets. J Biol Chem 2002; 277: 47588-95.
11 CAPRIE Steering Committee. A randomized, blinded, triala of clopidogrel versus aspirin in patients at risk of ischaemic events (CAPRIE). Lancet 1996; 348: 1329-39.
12 Vorchheimer D, Badimon J, Fuster V. Platelet glycoprotein IIb/IIIa receptor antagonists in cardiovascular disease. JAMA 1999; 281: 1407-14.
13 Moussa I, Oetgen M, Roubin G. et al Effectivness of clopidogrel and aspirin versus ticlopidine and aspirin in preventing stent thrombosis after coronary stent implantation. Circulation 1999; 99: 2364-6.
14 Mishkel GJ, Aguirre FV, Ligon RW. et al Clopidogrel as adjunctive antiplatelet therapy during coronary stenting. J Am Coll Cardiol 1999; 34: 1884-90.
15 Lau WC, Gurbel PA, Watkins PB. et al Contribution of hepatic cytochrome P450 3A4 metabolic activity to the phenomenon of clopidogrel resistance. Circulation 2004; 109: 166-71.
16 Matetzky S, Shenkman B, Guetta V. et al Clopidogrel resistance is associated with increased risk of recurrent atherothrombotic events in patients with acute myocardial infarction. Circulation 2004; 109: 3171-5.
17 Nurden AT. Platelet glycoprotein IIIa polymorphism and coronary thrombosis. Lancet 1997; 350: 1189-91.
18 Michelson AD, Furman MI, Goldschmidt-Clermont P. et al Platelet GP IIIa Pl(A) polymorphisms display different sensitivities to agonists. Circulation 2000; 101: 1013-8.
19 Feng D, Lindpaintner K, Larson MG. et al Increased platelet aggregability associated with platelet GPIIIa PlA2 polymorphism: the Framingham Offspring Study. Arterioscler Thromb Vasc Biol 1999; 19: 1142-7.
20 Undas A, Sanak M, Musial J. et al Platelet glycoprotein IIIa polymorphism, aspirin, and thrombin generation. Lancet 1999; 353: 982-3.
21 Szczeklik A, Undas A, Sanak M. et al Relationship between bleeding time, aspirin, and the PlA1/A2 polymorphism of platelet glycoprotein IIIa. Br J Haematol 2000; 110: 965-7.
22 Szczeklik A, Krzanowski M, Góra P. et al Antiplatelet drugs and generation of thrombin in clotting blood. Blood 1992; 80: 2006-11.
23 Sramek R, Sramek A, Koster T. et al A randomized and blinded comparison of three bleeding time techniques: the Ivy method, and the Simplate II method in two directions. Thromb Haemost 1992; 67: 514-8.
24 Schmitz G, Rothe G, Ruf A. et al European Working Group on Clinical Cell Analysis: Consensus protocol for the flow cytometric characterisation of platelet function. Thromb Haemost 1998; 79: 885-96.
25 Michelson AD, Bernard MR, Krueger LA. et al Evaluation of platelet function by flow cytometry. Methods 2000; 21: 259-70.
26 Angiolillo DJ, Fernandez-Ortiz A, Bernardo E. et al PlA polymorphism and platelet reactivity following clopidogrel loading dose in patients undergoing coronary stent implantation. Blood Coagul Fibrinolysis 2004; 15: 89-93.
27 Goodall AH, Curzen N, Panesar M. et al Increased binding of fibrinogen to glycoprotein IIIa-proline33 (HPA-1b, PlA2, Zwb) positive platelets in patients with cardiovascular disease. Eur Heart J 1999; 20: 742-7.
28 Vijayan KV, Goldschmidt-Clermont PJ, Roos C. et al The PlA2 polymorphism of integrin beta 3 enhances outside-in signaling and adhesive functions. J Clin Invest 2000; 105: 793-802.
29 Honda S, Honda Y, Bauer B. et al The impact of three-dimentional structure on the expression of PlA alloantigens on human integrin beta 3. Blood 1995; 86: 234-42.
30 Phillips JE, Lord ST, Gilbert GE. Fibrin stimulates platelets to increase factor VIIIa binding site expression. J Thromb Haemost 2004; 2: 1806-15.
31 Schober JM, Lam C-T, Wencel-Drake JD. Effect of cellular and receptor activation on the extent of integrin α_IIbβ₃ internalization. J Thromb Haemost 2003; 1: 2404-10.
32 Herault JP, Dol F, Gaich C. et al Effect of clopidogrel on thrombin generation in platelet-rich plasma in rat. Thromb Haemost 1999; 81: 957-60.
33 Roald HE, Barstad RM, Kierulf P. et al Clopidogrel-a platelet inhibitor which inhibits thrombogenesis in non-anticoagulated human blood independently of the blood flow conditions. Thromb Haemost 1994; 71: 655-62.
34 Shattil SJ, Kashiwagi H, Pampori M. Integrin signaling: the platelet paradigm. Blood 1998; 91: 2645-57.
35 Geiger J, Brich J, Hönig-Liedl P. et al Specific impairment of human platelet P2Yac ADP receptor-mediated signaling by the antiplatelet drug clopidogrel. Arterioscler Thromb Vasc Biol 1999; 19: 2007-11.
36 Furman MI, Krueger LA, Frelinger III AL. et al GPIIb-IIIa antagonist-induced reduction in platelet surface factor V/Va binding and phosphatydylserine expresion in whole blood. Thromb Haemost 2000; 84: 492-8.
37 Eikelboom JW, Weitz JI, Budaj A. et al Clopidogrel in unstable angina to prevent Recurrent Events (CURE) Study Investigators. Clopidogrel does not suppress blood markers of coagulation activation in aspirintreated patients with non-ST-elevation acute coronary syndromes. Eur Heart J 2002; 23: 1771-9.
38 Xiao T, Takagi J, Coller BS. et al Structural basis for allostery in integrins and binding to fibrinogen-mimetic therapeutics. Nature 2004; 432: 59-67.
39 Harrington RA, Becker BC, Ezekowitz M. et al Antithrombotic therapy for coronary artery disease. The seventh ACCP conference on thrombolytic therapy. Chest 2004; 126: 513S-548S.
40 Müller I, Besta F, Schulz C. et al Prevalence of clopidogrel non-responders among patients with stable angina pectoris scheduled for elective coronary stent placement. Thromb Haemost 2003; 89: 783-7.
41 CURE Trial Investigators. Effects of clopidogrel in addition to aspirin in patients with acute coronary syndromes without ST elevation. N Engl J Med 2001; 345: 494-502.
42 Undas A, Brummel K, Musial J. et al Pl^A2 Polymorphism of β₃ integrins is associated with enhanced thrombin generation and impaired antithrombotic action of aspirin at the site of microvascular injury. Circulation 2001; 104: 2666-72.
43 Hennekens CH, Schror K, Weisman S. et al Terms and conditions: semantic complexity and aspirin resistance. Circulation 2004; 110: 1706-8.