Download PDF
Thromb Haemost 2007; 97(03): 435-443
DOI: 10.1160/TH06-07-0415
Platelets and Blood Cells
Schattauer GmbH

Glycoprotein Ibα inhibition and ADP receptor antagonists, but not aspirin, reduce platelet thrombus formation in flowing blood exposed to atherosclerotic plaques

Sandra M. Penz
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
,
Armin J. Reininger
2   Department of Transfusion Medicine and Hemostaseology, University of Munich, Munich, Germany
,
Orsolya Toth
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
3   1stDepartment of Medicine, University of Pécs, Pécs, Hungary
,
Hans Deckmyn
4   Laboratory for Thrombosis Research, IRC, KU Leuven Campus Kortrijk, Kortrijk, Belgium
,
Richard Brandi
5   Department of Vascular Surgery, Klinikum Munich-Schwabing, Munich, Germany
,
Wolfgang Siess
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
› Author AffiliationsFinancial support: The study was supported by the grants from the Deutsche Forschungsgemeinschaft (Graduate Program GK 438 „Vascular Biology in Medicine”; Si-274/9) (S.M.P.; W.S.), the Friedrich-Baur foundation and the August-Lenz foundation (O.T).
Further Information

Publication History

Received 29 July 2006

Accepted after resubmission 29 February 2006

Publication Date:
28 November 2017 (online)

Also available at
    Permissions and Reprints

Summary

Anti-platelet drugs are used to prevent intra-arterial thrombus formation after rupture of atherosclerotic plaques. Until now, the inhibitory effect of present and future anti-platelet drugs such as aspirin, ADP receptor P2Y1/P2Y12 antagonists and glycoprotein (GP) Ibα inhibitors on the interaction of platelets with human plaques is not known. To study those effects we obtained human atherosclerotic plaques by surgical endarterectomy. Plaques induced maximal platelet aggregation in hirudinized platelet-rich plasma (PRP) and blood that was effectively inhibited by aspirin, the P2Y1 antagonist MRS2179 and the P2Y12 antagonist AR-C69931 MX, but not by GPIbα blockade with the mAB 6B4. Inhibition of platelet aggregation by MRS2179 was 74 ± 37% and 68 ± 20%, by AR-C69931 MX 94 ± 7% and 80 ± 6%, and by aspirin 88 ± 19% and 64 ± 28%, in PRP and blood, respectively (mean ± SD; n6–12 with plaques from 6 patients). The combination of both ADP receptor antagonists completely inhibited plaque-induced platelet aggregation in hirudinized PRP and blood. Under arterial flow conditions (1,500s-1), blockade of platelet GPIbα– resulted in a strong decrease of plaque-stimulated platelet adhesion/aggregate formation of 77 ± 5% (mean ± SD; n=4). Furthermore, MRS2I79, AR-C6993IMX and their combination reduced plaque-dependent platelet aggregate formation by 35 ± 14%, 32 ± 13% and 58 ± 12% (mean ± SD; n=5), respectively. Aspirin was without significant effect. In conclusion, a GPIbα-blocking antibody, as well as P2Y1 and P2Y12 receptor antagonists, alone or in combination, reduce in contrast to aspirin human plaque-induced platelet thrombus formation under arterial flow. Although these new anti-platelet agents inhibit platelet thrombus formation after plaque rupture, more efficient platelet blockers are required.

Keywords

ADP receptor antagonists - aspirin - GPIb - plaque - platelet throm bus formation
 

  • References

  • 1 Ruggeri ZM. Platelets in atherothrombosis. Nat Med 2002; 8: 1227-1234.
    CrossrefPubMedGoogle Scholar
  • 2 Corti R, Farkouh ME, Badimon JJ. The vulnerable plaque and acute coronary syndromes. Am J Med 2002; 113: 668-680.
    CrossrefPubMedGoogle Scholar
  • 3 Fernandez-Ortiz A, Badimon JJ, Falk E. et al. Characterization of the relative thrombogenicity of atherosclerotic plaque components: implications for consequences of plaque rupture. J Am Coll Cardiol 1994; 23: 1562-1569.
    CrossrefPubMedGoogle Scholar
  • 4 van Zanten GH, de Graaf S, Slootweg PJ. et al. Increased platelet deposition on atherosclerotic coronary arteries. J Clin Invest 1994; 93: 615-632.
    CrossrefPubMedGoogle Scholar
  • 5 Libby P. Inflammation in atherosclerosis. Nature 2002; 420: 868-874.
    CrossrefPubMedGoogle Scholar
  • 6 Siess W, Zangl KJ, Essler M. et al. Lysophosphatidic acid mediates the rapid activation of platelets and endothelial cells by mildly oxidized low density lipoprotein and accumulates in human atherosclerotic lesions. Proc Natl Acad Sci USA 1999; 96: 6931-6936.
    CrossrefPubMedGoogle Scholar
  • 7 Rother E, Brandl R, Baker DL. et al. Subtype-selective antagonists of lysophosphatidic Acid receptors inhibit platelet activation triggered by the lipid core of atherosclerotic plaques. Circulation 2003; 108: 741-747.
    CrossrefPubMedGoogle Scholar
  • 8 Penz S, Reininger AJ, Brandl R. et al. Human atheromatous plaques stimulate thrombus formation by activating platelet glycoprotein VI. FASEB J 2005; 19: 898-909.
    CrossrefPubMedGoogle Scholar
  • 9 Guyton JR, Klemp KF. Development of the lipidrich core in human atherosclerosis. Arterioscler Thromb Vasc Biol 1996; 16: 4-11.
    CrossrefPubMedGoogle Scholar
  • 10 Katsuda S, Kaji T. Atherosclerosis and extracellular matrix. J Atheroscler Thromb 2003; 10: 267-274.
    CrossrefPubMedGoogle Scholar
  • 11 De Meyer GR, Hoylaerts MF, Kockx MM. et al. Intimal deposition of functional von Willebrand factor in atherogenesis. Arterioscler Thromb Vasc Biol 1999; 19: 2524-2534.
    CrossrefPubMedGoogle Scholar
  • 12 Barnes MJ, Farndale RW. Collagens and atherosclerosis. Exp Gerontol 1999; 34: 513-525.
    CrossrefPubMedGoogle Scholar
  • 13 Abi-Younes S, Sauty A, Mach F. et al. The stromal cell-derived factor-1 chemokine is a potent platelet agonist highly expressed in atherosclerotic plaques. Circ Res 2000; 86: 131-138.
    CrossrefPubMedGoogle Scholar
  • 14 Weidtmann A, Scheithe R, Hrboticky N. et al. Mildly oxidized LDL induces platelet aggregation through activation of phospholipase A2. Arterioscler Thromb Vasc Biol 1995; 15: 1131-1138.
    CrossrefPubMedGoogle Scholar
  • 15 Merten M, Dong JF, Lopez JA. et al. Cholesterol sulfate: a new adhesive molecule for platelets. Circulation 2001; 103: 2032-2034.
    CrossrefPubMedGoogle Scholar
  • 16 Haseruck N, Erl W, Pandey D. et al. The plaque lipid lysophosphatidic acid stimulates platelet activation and platelet-monocyte aggregate formation in whole blood: involvement of P2Y1 and P2Y12 receptors. Blood 2004; 103: 2585-2592.
    CrossrefPubMedGoogle Scholar
  • 17 Ross R. Atherosclerosis--an inflammatory disease. N Engl J Med 1999; 340: 115-126.
    CrossrefPubMedGoogle Scholar
  • 18 Kariyazono H, Nakamura K, Arima J. et al. Evaluation of anti-platelet aggregatory effects of aspirin, cilostazol and ramatroban on platelet-rich plasma and whole blood. Blood Coagul Fibrinolysis 2004; 15: 157-167.
    CrossrefPubMedGoogle Scholar
  • 19 Phillips DR, Conley PB, Sinha U. et al. Therapeutic approaches in arterial thrombosis. J Thromb Haemost 2005; 3: 1577-1589.
    CrossrefPubMedGoogle Scholar
  • 20 Barstad RM, Orvim U, Hamers MJ. et al. Reduced effect of aspirin on thrombus formation at high shear and disturbed laminar blood flow. Thromb Haemost 1996; 75: 827-832.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Eikelboom JW, Hankey GJ. Failure of aspirin to prevent atherothrombosis: potential mechanisms and implications for clinical practice. Am J Cardiovasc Drugs 2004; 4: 57-67.
    CrossrefPubMedGoogle Scholar
  • 22 Hanson SR, Sakariassen KS. Blood flow and antithrombotic drug effects. Am Heart J 1998; 135: S132-145.
    CrossrefPubMedGoogle Scholar
  • 23 Cosemans JM, Kuijpers MJ, Lecut C. et al. Contribution of platelet glycoprotein VI to the thrombogenic effect of collagens in fibrous atherosclerotic lesions. Atherosclerosis 2005; 181: 19-27.
    CrossrefPubMedGoogle Scholar
  • 24 Toth O, Calatzis A, Penz S. et al. Multiple electrode aggregometry: A new device to measure platelet aggregation in whole blood. Thromb Haemost 2006; 96: 781-788.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Cauwenberghs N, Vanhoorelbeke K, Vauterin S. et al. Epitope mapping of inhibitory antibodies against platelet glycoprotein Ibalpha reveals interaction between the leucine-rich repeat N-terminal and C-terminal flanking domains of glycoprotein Ibalpha. Blood 2001; 98: 652-660.
    CrossrefPubMedGoogle Scholar
  • 26 Brandl R, Richter T, Haug K. et al. Topographic analysis of proliferative activity in carotid endarterectomy specimens by immunocytochemical detection of the cell cycle-related antigen Ki-67. Circulation 1997; 96: 3360-3368.
    CrossrefPubMedGoogle Scholar
  • 27 Savi P, Bernat A, Lale A. et al. Effect of aspirin on platelet desaggregation induced by SR121566, a potent GP-IIb/IIIa antagonist. Platelets 2000; 11: 43-48.
    CrossrefPubMedGoogle Scholar
  • 28 Cullen P, Baetta R, Bellosta S. et al. Rupture of the atherosclerotic plaque: Does a good animal model exist?. Arterioscler Thromb Vasc Biol 2003; 23: 529-534.
    CrossrefPubMedGoogle Scholar
  • 29 Schumacher KA, Classen HG, Spath M. Platelet aggregation evoked in vitro and in vivo by phosphatidic acids and lysoderivatives: identity with substances in aged serum (DAS). Thromb Haemost 1979; 42: 631-640.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Gerrard JM, Kindom SE, Peterson DA. et al. Lysophosphatidic acids. II. Interaction of the effects of adenosine diphosphate and lysophosphatidic acids in dog, rabbit, and human platelets. Am J Pathol 1979; 97: 531-547.
    PubMedGoogle Scholar
  • 31 Tokamura A, Fukuzawa K, Isobe J. et al. Lysophosphatidic acid-induced aggregation of human and feline platelets: structure-activity relationship. Biochem Biophys Res Commun 1981; 99: 391-398.
    CrossrefPubMedGoogle Scholar
  • 32 Belton OA, Duffy A, Toomey S. et al. Cyclooxygenase isoforms and platelet vessel wall interactions in the apolipoprotein E knockout mouse model of atherosclerosis. Circulation 2003; 108: 3017-3023.
    CrossrefPubMedGoogle Scholar
  • 33 Gawaz M. Platelets in the onset of atherosclerosis. Blood Cells Mol Dis 2006; 36: 206-210.
    CrossrefPubMedGoogle Scholar
  • 34 Savage B, Saldivar E, Ruggeri ZM. Initiation of platelet adhesion by arrest onto fibrinogen or translocation on von Willebrand factor. Cell 1996; 84: 289-297.
    CrossrefPubMedGoogle Scholar
  • 35 Savage B, Almus-Jacobs F, Ruggeri ZM. Specific synergy of multiple substrate-receptor interactions in platelet thrombus formation under flow. Cell 1998; 94: 657-666.
    CrossrefPubMedGoogle Scholar
  • 36 Sugimoto M, Tsuji S, Kuwahara M. et al. Shear-dependent functions of the interaction between soluble von Willebrand factor and platelet glycoprotein Ib in mural thrombus formation on collagen surface. Int J Hematol 1999; 69: 48-53.
    PubMedGoogle Scholar
  • 37 Maxwell MJ, Westein E, Nesbitt WS. et al. Identification of a two-stage platelet aggregation process mediating shear-dependent thrombus formation. Blood. 2006 epub ahead of print
    PubMedGoogle Scholar
  • 38 Ruggeri ZM, Dent JA, Saldivar E. Contribution of distinct adhesive interactions to platelet aggregation in flowing blood. Blood 1999; 94: 172-178.
    PubMedGoogle Scholar
  • 39 Ruggeri ZM, Orje JN, Habermann R. et al. Activation- independent platelet adhesion and aggregation under elevated shear stress. Blood 2006; 108: 1903-1910.
    CrossrefPubMedGoogle Scholar
  • 40 Kunapuli SP, Dorsam RT, Kim S. et al. Platelet purinergic receptors. Curr Opin Pharmacol 2003; 3: 175-180.
    CrossrefPubMedGoogle Scholar
  • 41 Turner NA, Moake JL, McIntire LV. Blockade of adenosine diphosphate receptors P2Y(12) and P2Y(1) is required to inhibit platelet aggregation in whole blood under flow. Blood 2001; 98: 3340-3345.
    CrossrefPubMedGoogle Scholar
  • 42 Moake JL, Turner NA, Stathopoulos NA. et al. Shear-induced platelet aggregation can be mediated by vWF released from platelets, as well as by exogenous large or unusually large vWF multimers, requires adenosine diphosphate, and is resistant to aspirin. Blood 1988; 71: 1366-1374.
    PubMedGoogle Scholar
  • 43 Gachet C. The platelet P2 receptors as molecular targets for old and new antiplatelet drugs. Pharmacol Ther 2005; 108: 180-192.
    CrossrefPubMedGoogle Scholar
  • 44 Vilahur G, Segales E, Salas E. et al. Effects of a novel platelet nitric oxide donor (LA816), aspirin, clopidogrel, and combined therapy in inhibiting flowand lesion-dependent thrombosis in the porcine ex vivo model. Circulation 2004; 110: 1686-1693.
    CrossrefPubMedGoogle Scholar
  • 45 Remijn JA, Wu YP, Jeninga EH. et al. Role of ADP receptor P2Y(12) in platelet adhesion and thrombus formation in flowing blood. Arterioscler Thromb Vasc Biol 2002; 22: 686-691.
    CrossrefPubMedGoogle Scholar
  • 46 Goto S, Tamura N, Ishida H. et al. Dependence of platelet thrombus stability on sustained glycoprotein IIb/IIIa activation through adenosine 5'-diphosphate receptor stimulation and cyclic calcium signaling. J Am Coll Cardiol 2006; 47: 155-162.
    CrossrefPubMedGoogle Scholar
  • 47 Goto S, Tamura N, Handa S. Effects of adenosine 5'-diphosphate (ADP) receptor blockade on platelet aggregation under flow. Blood 2002; 99: 4644-4645.
    CrossrefPubMedGoogle Scholar