Skip to main content

Advertisement

SpringerLink
Log in

A platelet target for venous thrombosis? P2Y1 deletion or antagonism protects mice from vena cava thrombosis

  • Published:
Journal of Thrombosis and Thrombolysis Aims and scope Submit manuscript

Abstract

A role for platelets in the pathogenesis of venous thrombosis was suggested by clinical and preclinical studies. However, examination of the platelet receptor, P2Y1, in this area has been limited. The goal of the current study was to examine effects of P2Y1 deletion, or selective antagonism with MRS2500, in oxidative venous thrombosis in mice. The P2Y12 antagonist, clopidogrel, was included as a reference agent. Anesthetized C57BL/6 or genetically modified mice underwent 3.5 or 5 % FeCl3-induced vena cava thrombosis. Pharmacokinetic properties of MRS2500 were defined for dose selection. Platelet aggregation and renal or tail bleeding times (BT) were measured to put antithrombotic effects into perspective. P2Y1 deletion significantly reduced (p < 0.001) venous thrombus weight by 74 % in 3.5 % FeCl3 injury compared to P2Y1+/+ littermates. MRS2500 (2 mg/kg, i.v.) significantly decreased (p < 0.001) thrombus weight 64 % in C57BL/6 mice. In the more severe 5 % FeCl3-induced injury model, thrombus weight significantly (p < 0.001) decreased 68 % in P2Y1−/− mice versus P2Y1+/+ mice, and MRS2500 (2 mg/kg) was also beneficial (54 % decrease, p < 0.01). Renal BT doubled in P2Y1−/− versus P2Y1+/+ mice, and increased threefold with MRS2500 compared to vehicle. Tail BT was markedly prolonged in P2Y1−/− mice (7.9X) and in C57BL/6 mice given MRS2500. The current study demonstrates that P2Y1 deletion or antagonism significantly reduced venous thrombosis in mice, suggesting that P2Y1 receptors play a role in the pathogenesis of venous thrombosis, at least in this species. However as with many antithrombotic agents the benefit comes at the potential price of an increase in provoked bleeding times.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

BT:

Bleeding time

ADP:

Adenosine 5′-diphosphate

cAMP:

Cyclic adenosine monophosphate

MeOH:

Methanol

LC/MS:

Liquid chromatography/mass spectrometry

i.v.:

Intravenous

i.p.:

Intraperitoneal

References

  1. Martinelli I, Bucciarelli P, Mannucci PM (2010) Thrombotic risk factors: basic pathophysiology. Crit Care Med 38(2):S3–S9

    Article  PubMed  Google Scholar 

  2. Brill A, Fuchs TA, Chauhan AK, Yang JJ, De Meyer SF, Kollnberger M, Wakefield TW, Lammle B, Massberg S, Wagner DD (2011) von Willebrand factor–mediated platelet adhesion is critical for deep vein thrombosis in mouse models. Blood 117:1400–1407

    Article  PubMed  CAS  Google Scholar 

  3. Schreijer AJM, Hoylaerts MF, Meijers JCM, Lijnen HR, Middeldorp S, Buller HR, Reitsma PH, Rosendaal FR, Cannegieter SC (2010) Explanations for coagulation activation after air travel. J Thromb Haemost 8:971–978

    PubMed  CAS  Google Scholar 

  4. Leoncini G, Signorello MG, Segantin A, Giacobbe E, Armani U, Piana A, Camicione P (2009) In retinal vein occlusion platelet response to thrombin is increased. Thromb Res 124:e48–e55

    Article  PubMed  CAS  Google Scholar 

  5. Kageyama K, Nakajima Y, Shibasaki M, Hashimoto S, Mizobe T (2007) Increased platelet, leukocyte, and endothelial cell activity are associated with increased coagulability in patients after total knee arthroplasty. J Thromb Haemost 5:738–745

    Article  PubMed  CAS  Google Scholar 

  6. Leon C, Alex M, Klocke A, Morgenstern E, Moosbauer C, Eckly A, Spannagl M, Gachet C, Engelmann B (2004) Platelet ADP receptors contribute to the initiation of intravascular coagulation. Blood 103:594–600

    Article  PubMed  CAS  Google Scholar 

  7. Gachet C, Hechler B (2005) The platelet P2 receptors in thrombosis. Semin Thromb Hemost 31:162–167

    Article  PubMed  CAS  Google Scholar 

  8. Hechler B, Nonne C, Roh EJ, Cattaneo M, Cazenave J-P, Lanza F, Jacobson KA, Gachet C (2006) MRS2500 [2-iodo-N6-methyl-(N)-methanocarba-2′-deoxyadenosine-3′,5′-bisphosphate], a potent, selective, and stable antagonist of the platelet P2Y1 receptor with strong antithrombotic activity in mice. J Pharmacol Exp Ther 316:556–563

    Article  PubMed  CAS  Google Scholar 

  9. Gachet C (2006) Regulation of platelet functions by P2 receptors. Annu Rev Pharmacol Toxicol 46:277–300

    Article  PubMed  CAS  Google Scholar 

  10. Fabre JE, Nguyen M, Latour A, Keifer JA, Audoly LP, Coffman TM, Koller BH (1999) Decreased platelet aggregation, increased bleeding time and resistance to thromboembolism in P2Y1-deficient mice. Nat Med 5:1199–1202

    Article  PubMed  CAS  Google Scholar 

  11. Léon C, Hechler B, Freund M, Eckly A, Vial C, Ohlmann P, Dierich A, LeMeur M, Cazenave JP, Gachet C (1999) Defective platelet aggregation and increased resistance to thrombosis in purinergic P2Y(1) receptor-null mice. J Clin Invest 104:1731–1737

    Article  PubMed  Google Scholar 

  12. Léon C, Freund M, Ravanat C, Baurand A, Cazenave JP, Gachet C (2001) Key role of the P2Y(1) receptor in tissue factor-induced thrombin-dependent acute thromboembolism: studies in P2Y(1)-knockout mice and mice treated with a P2Y(1) antagonist. Circulation 103:718–723

    Article  PubMed  Google Scholar 

  13. Hechler B, Gachet C (2011) P2 receptors and platelet function. Purinergic Signal 7:293–303

    Article  PubMed  CAS  Google Scholar 

  14. Zerr M, Hechler B, Freund M, Magnenat S, Lanois I, Cazenave J-P, Leon C, Gachet C (2011) Major contribution of the P2Y1 receptor in purinergic regulation of TNFα-induced vascular inflammation. Circulation 123:2404–2413

    Article  PubMed  CAS  Google Scholar 

  15. Laplante M-A, Monassier L, Freund M, Bousquet P, Gachet C (2010) The purinergic P2Y1 receptor supports leptin secretion in adipose tissue. Endocrinology 151:2060–2070

    Article  PubMed  CAS  Google Scholar 

  16. Hechler B, Freund M, Ravanat C, Magnenat S, Cazenave J-P, Gachet C (2008) Reduced atherosclerotic lesions in P2Y1/apolipoprotein E double-knockout mice. Circulation 118:754–763

    Article  PubMed  CAS  Google Scholar 

  17. Lenain N, Freund M, Léon C, Cazenave JP, Gachet C (2003) Inhibition of localized thrombosis in P2Y1-deficient mice and rodents treated with MRS2179, a P2Y1 receptor antagonist. J Thromb Hemost 1:1144–1149

    Article  CAS  Google Scholar 

  18. Kim HS, Ohno M, Xu B, Kim HO, Choi Y, Ji XD, Maddileti S, Marquez VE, Harden TK, Jacobson KA (2003) 2-Substitution of adenine nucleotide analogues containing a bicyclo[3.1.0]hexane ring system locked in a northern conformation: enhanced potency as P2Y1 receptor antagonists. J Med Chem 46:4974–4987

    Article  PubMed  CAS  Google Scholar 

  19. Wang X, Smith PL, Hsu M-Y, Gailani D, Schumacher WA, Ogletree ML (2006) Effects of factor XI deficiency on ferric chloride-induced vena cava thrombosis in mice. J Thromb Haemost 4:1982–1988

    Article  PubMed  CAS  Google Scholar 

  20. Schumacher WA, Bostwick JS, Ogletree ML, Stewart A, Steinbacher TE, Hua J, Price L, Wong PC, Rehfuss R (2007) Biomarker optimization to track the antithrombotic and hemostatic effects of clopidogrel in rats. J Pharmacol Exp Ther 322:1–9

    Article  Google Scholar 

  21. Cattaneo M, Lecchi A, Ohno M, Joshi BV, Besada P, Tchilibon S, Lombardi R, Bischofberger N, Harden TK, Jacobson KA (2004) Antiaggregatory activity in human platelets of potent antagonists of the P2Y 1 receptor. Biochem Pharmacol 68:1995–2002

    Article  PubMed  CAS  Google Scholar 

  22. Ohlmann P, de Castro S, Brown GG, Gachet C, Jacobson KA, Harden TK (2010) Quantification of recombinant and platelet P2Y1 receptors utilizing a [125I]-labeled high-affinity antagonist 2-iodo-N6-methyl-(N)-methanocarba-2′-deoxyadenosine-3′,5′-bisphosphate ([125I]MRS2500. Pharmacol Res 62:344–351

    Article  PubMed  CAS  Google Scholar 

  23. Kahner BN, Dorsam RT, Kunapuli SP (2008) Role of P2Y receptor subtypes in platelet-derived microparticle generation. Front Biosci 13:433–439

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank George Psaltis and Amy Saari for breeding and caring for the P2Y1 knockout mice, Glenn Cantor for helping to establish congenic analysis of mice, and Jinwen Huang for genotyping of mice. The authors thank Carl Thibault and Rejean Ruel for synthesis of MRS2500, and Robert Rehfuss and Pancras Wong for helpful advice.

Conflict of interest

All authors were employees of Bristol-Myers Squibb at the time these studies were performed.

Author information

Authors and Affiliations

  1. Department of Thrombosis Biology, Bristol-Myers Squibb, 311 Pennington-Rocky Hill Road, Pennington, NJ, 08534, USA

    J. Eileen Bird, Patricia L. Smith, Frank Barbera, Christine Huang & William A. Schumacher

  2. Agennix, Inc, 101 College Road East, Princeton, NJ, 08540, USA

    Xinkang Wang

Authors
  1. J. Eileen Bird
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xinkang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Patricia L. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Frank Barbera
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christine Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. William A. Schumacher
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J. Eileen Bird.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bird, J.E., Wang, X., Smith, P.L. et al. A platelet target for venous thrombosis? P2Y1 deletion or antagonism protects mice from vena cava thrombosis. J Thromb Thrombolysis 34, 199–207 (2012). https://doi.org/10.1007/s11239-012-0745-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11239-012-0745-3

Keywords

Search

Navigation