The influence of variation in the P2Y12 receptor gene on in vitro platelet inhibition with the direct P2Y12 antagonist cangrelor

Heleen J. Bouman; Jochem W. van Werkum; Goran Rude; Frank W. G. Leebeek; Adrian Kruit; Christian M. Hackeng; Jurriën M. ten Berg; Moniek P. M. de Maat; Henk J. T. Ruven

doi:10.1160/TH09-06-0367

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2010; 103(02): 379-386
DOI: 10.1160/TH09-06-0367

Platelets and Blood Cells

Schattauer GmbH

The influence of variation in the P2Y12 receptor gene on in vitro platelet inhibition with the direct P2Y12 antagonist cangrelor

Heleen J. Bouman

¹Department of Cardiology, St. Antonius Hospital Nieuwegein, the Netherlands

,

Jochem W. van Werkum

¹Department of Cardiology, St. Antonius Hospital Nieuwegein, the Netherlands

,

Goran Rude

²Department of Hematology, Erasmus University Medical Center Rotterdam, the Netherlands

,

Frank W. G. Leebeek

²Department of Hematology, Erasmus University Medical Center Rotterdam, the Netherlands

,

Adrian Kruit

³Department of Clinical Chemistry, St. Antonius Hospital Nieuwegein, the Netherlands

,

Christian M. Hackeng

³Department of Clinical Chemistry, St. Antonius Hospital Nieuwegein, the Netherlands

,

Jurriën M. ten Berg

¹Department of Cardiology, St. Antonius Hospital Nieuwegein, the Netherlands

,

Moniek P. M. de Maat

²Department of Hematology, Erasmus University Medical Center Rotterdam, the Netherlands

,

Henk J. T. Ruven

³Department of Clinical Chemistry, St. Antonius Hospital Nieuwegein, the Netherlands

› Author Affiliations

Abstract

Summary

Novel P2Y12 inhibitors are in development to overcome the occurrence of atherothrombotic events associated with poor responsiveness to the widely used P2Y12 inhibitor clopidogrel. Cangrelor is an intravenously administered P2Y12 inhibitor that does not need metabolic conversion to an active metabolite for its antiplatelet action, and as a consequence exhibits a more potent and consistent antiplatelet profile as compared to clopidogrel. It was the objective of this study to determine the contribution of variation in the P2Y12 receptor gene to platelet aggregation after in vitro partial P2Y12 receptor blockade with the direct antagonist cangrelor. Optical aggregometry was performed at baseline and after in vitro addition of 0.05 and 0.25 μM cangrelor to the platelet-rich plasma of 254 healthy subjects. Five haplotype-tagging (ht)-SNPs covering the entire P2Y12 receptor gene were genotyped (rs6798347C>t, rs6787801T>c, rs9859552C>a, rs6801273A>g and rs2046934T>c [T744C]) and haplotypes were inferred. The minor c allele of SNP rs6787801 was associated with a 5% lower 20 μM ADP-induced peak platelet aggregation (0.05 μM cangrelor, p<0.05). Aa homozygotes for SNP rs9859552 showed 20% and 17% less inhibition of platelet aggregation with cangrelor when compared to CC homozygotes (0.05 and 0.25 μM cangrelor respectively; p<0.05). Results of the haplotype analyses were consistent with those of the single SNPs. Polymorphisms of the P2Y12 receptor gene contribute significantly to the interindividual variability in platelet inhibition after partial in vitro blockade with the P2Y12 antagonist cangrelor.

Keywords

Cangrelor - P2Y12 receptor - single nucleotide polymorphism - haplotypes - platelet aggregation

Full Text

References

References
1 Serebruany VL, Steinhubl SR, Berger PB. et al. Variability in platelet responsiveness to clopidogrel among 544 individuals. J Am Coll Cardiol 2005; 45: 246-251.
2 Gurbel PA, Bliden KP, Hiatt BL. et al. Clopidogrel for coronary stenting: response variability, drug resistance, and the effect of pretreatment platelet reactivity. Circulation 2003; 107: 2908-2913.
3 Taubert D, Kastrati A, Harlfinger S. et al. Pharmacokinetics of clopidogrel after administration of a high loading dose. Thromb Haemost 2004; 92: 311-316.
4 Taubert D, von Beckerath N, Grimberg G. et al. Impact of P-glycoprotein on clopidogrel absorption. J Pharmacol Exp Ther 2006; 80: 486-501.
5 Michelson AD, Linden MD, Furman MI. et al. Evidence that pre-existent variability in platelet response to ADP accounts for “clopidogrel resistance“. J Thromb Haemost 2007; 5: 75-81.
6 Trenk D, Hochholzer W, Fromm MF. et al. Cytochrome P450 2C19 681G>A polymorphism and high on-clopidogrel platelet reactivity associated with adverse 1-year clinical outcome of elective percutaneous coronary intervention with drug-eluting or bare-metal stents. J Am Coll Cardiol 2008; 51: 1925-1934.
7 Mega JL, Close SL, Wiviott SD. et al. Cytochrome p-450 polymorphisms and response to clopidogrel. N Engl J Med 2009; 360: 354-362.
8 Hulot JS, Bura A, Villard E. et al. Cytochrome P450 2C19 loss-of-function polymorphism is a major determinant of clopidogrel responsiveness in healthy subjects. Blood 2006; 108: 2244-2247.
9 van Werkum JW, Heestermans AACM, Deneer VHM. et al. Clopidogrel resistance: Fact and Fiction. Future Cardiology 2006; 2: 215-228.
10 Fontana P, Dupont A, Gandrille S. et al. Adenosine diphosphate-induced platelet aggregation is associated with P2Y₁₂ gene sequence variations in healthy subjects. Circulation 2003; 108: 989-995.
11 Staritz P, Kurz K, Stoll M. et al. Platelet reactivity and clopidogrel resistance are associated with the H2 haplotype of the P2Y(12)-ADP receptor gene. Int J Cardiol 2009; 133: 341-345.
12 Evans DJ, Jackman LE, Chamberlain J. et al. Platelet P2Y(12) receptor influences the vessel wall response to arterial injury and thrombosis. Circulation 2009; 119: 116-122.
13 Michelson AD. P2Y₁₂ Antagonism. Promises and Challenges. Arterioscler Thromb Vasc Biol 2008; 28: s33-38.
14 Storey RF, Wilcox RG, Heptinstall S. Comparison of the pharmacodynamic effects of the platelet ADP receptor antagonists clopidogrel and AR-C69931MX in patients with ischaemic heart disease. Platelets 2002; 13: 407-413.
15 Fugate SE, Cudd LA. Cangrelor for treatment of coronary thrombosis. Ann Pharmacother 2006; 40: 925-930.
16 A Clinical Trial to Demonstrate the Efficacy of Cangrelor (CHAMPION-PCI). Available at http://www.clinicaltrials.gov/ct2/show/NCT00305162?term=cangrelor&rank=4. Accessed April 2, 2009.
17 van Werkum JW, Kleibeuker M, Mieremet N. et al. Evaluation of the platelet response to clopidogrel with light transmittance aggregometry: peak aggregation or late aggregation?. J Thromb Haemost 2007; 5: 884-886.
18 Rudez G, Pons D, Leebeek F. Platelet receptor P2RY12 haplotypes predict restenosis after percutaneous coronary interventions. Hum Mutat 2008; 29: 375-380.
19 Livak KJ. Allelic discrimination using fluorogenic probes and the 5’ nuclease assay. Genet Anal 1999; 14: 143-149.
20 Hetherington SL, Singh RK, Lodwick D. et al. Dimorphism in the P2Y1 ADP receptor gene is associated with increased platelet activation response to ADP. Arterioscler Thromb Vasc Biol 2005; 25: 252-257.
21 Giusti B, Gori AM, Marcucci R. et al. Cytochrome P450 2C19 loss-of-function polymorphism, but not CYP3A4 IVS10 + 12G/A and P2Y₁₂ T744C polymorphisms, is associated with response variability to dual antiplatelet treatment in high-risk vascular patients. Pharmacogenet Genomics 2007; 17: 1057-1064.
22 von Beckerath N, von Beckerath O, Koch W. et al. P2Y₁₂ gene H2 haplotype is not associated with increased adenosine diphosphate-induced platelet aggregation after initiation of clopidogrel therapy with a high loading dose. Blood Coagul Fibrinolysis 2005; 16: 199-204.
23 Angiolillo DJ, Fernandez-Ortiz A, Bernardo E. et al. Lack of association between the P2Y₁₂ receptor gene polymorphism and platelet response to clopidogrel in patients with coronary artery disease. Thromb Res 2005; 116: 491-497.
24 Lev EI, Patel RT, Guthikonda S. et al. Genetic polymorphisms of the platelet receptors P2Y(12), P2Y(1) and GP IIIa and response to aspirin and clopidogrel. Thromb Res 2007; 119: 355-360.
25 Bura A, Bachelot-Loza C, Ali FD. et al. Role of the P2Y₁₂ gene polymorphism in platelet responsiveness to clopidogrel in healthy subjects. J Thromb Haemost 2006; 4: 2096-2097.
26 Bernardo E, Angiolillo DJ, Ramirez C. et al. Lack of association between gene sequence variations of platelet membrane receptors and aspirin responsiveness detected by the PFA-100 system in patients with coronary artery disease. Platelets 2006; 17: 586-590.
27 Cuisset T, Frere C, Quilici J. et al. Role of the T744C polymorphism of the P2Y₁₂ gene on platelet response to a 600-mg loading dose of clopidogrel in 597 patients with non-ST-segment elevation acute coronary syndrome. Thromb Res 2007; 120: 893-899.
28 De Miguel A, Ibanez B, Badimon JJ. Clinical implications of clopidogrel resistance. Thromb Haemost 2008; 100: 196-203.
29 Elsenberg EHAM, van Werkum JW, van de Wal RMA. et al. The influence of clinical characteristics, laboratory and inflammatory markers on ‘high on-treatment platelet reactivity’ as measured with different platelet function tests. Thromb Haemost 2009; 102: 719-727.
30 Geisler T, Langer H, Wydymus M. et al. Low response to clopidogrel is associated with cardiovascular outcome after coronary stent implantation. Eur Heart J 2006; 27: 2420-2425.
31 Hochholzer W, Trenk D, Bestehorn HP. et al. Impact of the degree of peri-inter -ventional platelet inhibition after loading with clopidogrel on early clinical outcome of elective coronary stent placement. J Am Coll Cardiol 2006; 48: 1742-1750.
32 Buonamici P, Marcucci R, Migliorini A. et al. Impact of platelet reactivity after clopidogrel administration on drug-eluting stent thrombosis. J Am Coll Cardiol 2007; 49: 2312-2317.
33 Price MJ, Endemann S, Gollapudi RR. et al. Prognostic significance of post-clopidogrel platelet reactivity assessed by a point-of-care assay on thrombotic events after drug-eluting stent implantation. Eur Heart J 2009; 29: 992-1000.
34 Wiviott SD, Braunwald E, McCabe CH. et al. Prasugrel versus clopidogrel in patients with acute coronary syndromes. N Engl J Med 2007; 357: 2001-2015.
35 Wiviott SD, Trenk D, Frelinger AL. et al. Prasugrel compared with high loading-and maintenance-dose clopidogrel in patients with planned percutaneous coronary intervention: the Prasugrel in Comparison to Clopidogrel for Inhibition of Platelet Activation and Aggregation-Thrombolysis in Myocardial Infarction 44 trial. Circulation 2007; 116: 2923-2932.
36 den Dunnen JT, Antonarakis SE. Mutation nomenclature extensions and suggestions to describe complex mutations: a discussion. Hum Mutat 2000; 15: 7-12.