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Thromb Haemost 2014; 111(03): 438-446
DOI: 10.1160/TH13-05-0360
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Influence of single nucleotide polymorphisms on thrombin generation in factor V Leiden heterozygotes

Olivier Segers
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Centre, Maastricht, the Netherlands
,
Paolo Simioni
2   Department of Cardiologic, Thoracic and Vascular Sciences, 2nd Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Daniela Tormene
2   Department of Cardiologic, Thoracic and Vascular Sciences, 2nd Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Elisabetta Castoldi
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Centre, Maastricht, the Netherlands
› Author Affiliations Financial support:This work was supported by a VIDI grant (nr. 917–76–312, to E. Castoldi) from the Dutch Organisation for Scientific Research (NWO).
Further Information

Publication History

Received: 02 May 2013

Accepted after major revision: 08 October 2013

Publication Date:
22 November 2017 (online)

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Summary

Carriership of the factor V (FV) Leiden mutation increases the risk of venous thromboembolism (VTE) ~4-fold, but the individual risk of each FV Leiden carrier depends on several co-inherited risk and protective factors. Under the hypothesis that thrombin generation might serve as an intermediate phenotype to identify genetic modulators of VTE risk, we enrolled 188 FV Leiden heterozygotes (11 with VTE) and determined the following parameters: thrombin generation in the absence and presence of activated protein C (APC); plasma levels of prothrombin, factor X, antithrombin, protein S and tissue factor pathway inhibitor; and the genotypes of 24 SNPs located in the genes encoding these coagulation factors and inhibitors. Multiple regression analysis was subsequently applied to identify the (genetic) determinants of thrombin generation. The endogenous thrombin potential (ETP) showed a striking inter-individual variability among different FV Leiden carriers and, especially when measured in the presence of APC, correlated with VTE risk. Several SNPs in the F2 (rs1799963, rs3136516), F10 (rs693335), SERPINC1 (rs2227589), PROS1 (Heerlen polymorphism) and TFPI (rs5940) genes significantly affected the ETPAPC and/or the ETP+APC in FV Leiden carriers. Most of these SNPs have shown an association with VTE risk in conventional epidemiological studies, suggesting that the genetic dissection of thrombin generation leads to the detection of clinically relevant SNPs. In conclusion, we have identified several SNPs that modulate thrombin generation in FV Leiden heterozygotes. These SNPs may help explain the large variability in VTE risk observed among different FV Leiden carriers.

Keywords

Factor V Leiden - intermediate phenotype - single nucleotide polymorphism - thrombin generation - venous thromboembolism

 

  • References

  • 1 Bertina RM, Koeleman BP, Koster T. et al. Mutation in blood coagulation factor V associated with resistance to activated protein C.. Nature 1994; 369: 64-67.
    CrossrefPubMedGoogle Scholar
  • 2 Simone B, De Stefano V, Leoncini E. et al. Risk of venous thromboembolism associated with single and combined effects of Factor V Leiden, Prothrombin 20210A and Methylenetethraydrofolate reductase C677T: a meta-analysis involving over 11,000 cases and 21,000 controls.. Eur J Epidemiol 2013; 28: 621-647.
    CrossrefPubMedGoogle Scholar
  • 3 Kalafatis M, Bertina RM, Rand MD. et al. Characterization of the molecular defect in factor VR506Q.. J Biol Chem 1995; 270: 4053-4057.
    CrossrefPubMedGoogle Scholar
  • 4 Nicolaes GA, Tans G, Thomassen MC. et al. Peptide bond cleavages and loss of functional activity during inactivation of factor Va and factor VaR506Q by activated protein C.. J Biol Chem 1995; 270: 21158-21166.
    CrossrefPubMedGoogle Scholar
  • 5 Thorelli E, Kaufman RJ, Dahlbäck B. Cleavage of factor V at Arg 506 by activated protein C and the expression of anticoagulant activity of factor V.. Blood 1999; 93: 2552-2558.
    PubMedGoogle Scholar
  • 6 Castoldi E, Brugge JM, Nicolaes GA. et al. Impaired APC cofactor activity of factor V plays a major role in the APC resistance associated with the factor V Leiden (R506Q) and R2 (H1299R) mutations.. Blood 2004; 103: 4173-4179.
    CrossrefPubMedGoogle Scholar
  • 7 Dahlbäck B, Carlsson M, Svensson PJ. Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C.. Proc Natl Acad Sci USA 1993; 90: 1004-1008.
    CrossrefPubMedGoogle Scholar
  • 8 Rodeghiero F, Tosetto A. Activated protein C resistance and factor V Leiden mutation are independent risk factors for venous thromboembolism.. Ann Intern Med 1999; 130: 643-650.
    CrossrefPubMedGoogle Scholar
  • 9 Heit JA, Sobell JL, Li H. et al. The incidence of venous thromboembolism among Factor V Leiden carriers: a community-based cohort study.. J Thromb Haemost 2005; 3: 305-311.
    CrossrefPubMedGoogle Scholar
  • 10 Rosendaal FR. Venous thrombosis: a multicausal disease.. Lancet 1999; 353: 1167-1173.
    CrossrefPubMedGoogle Scholar
  • 11 Simioni P, Castoldi E, Lunghi B. et al. An underestimated combination of oppo-sites resulting in enhanced thrombotic tendency.. Blood 2005; 106: 2363-2365.
    CrossrefPubMedGoogle Scholar
  • 12 Mingozzi F, Legnani C, Lunghi B. et al. A FV multiallelic marker detects genetic components of APC resistance contributing to venous thromboembolism in FV Leiden carriers.. Thromb Haemost 2003; 89: 983-989.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Segers O, Simioni P, Tormene D. et al. Genetic modulation of the FV Leiden/ normal FV ratio and risk of venous thrombosis in FV Leiden heterozygotes.. J Thromb Haemost 2012; 10: 73-80.
    CrossrefPubMedGoogle Scholar
  • 14 Tosetto A, Rodeghiero F, Martinelli I. et al. Additional genetic risk factors for venous thromboembolism in carriers of the factor V Leiden mutation.. Br J Hae-matol 1998; 103: 871-876.
    CrossrefPubMedGoogle Scholar
  • 15 Meinardi JR, Middeldorp S, de Kam PJ. et al. Risk of venous thromboembolism in carriers of factor V Leiden with a concomitant inherited thrombophilic defect: a retrospective analysis.. Blood Coagul Fibrinolysis 2001; 12: 713-720.
    CrossrefPubMedGoogle Scholar
  • 16 Medina P, Navarro S, Estellés A. et al. Influence of the 4600A/G and 4678G/C polymorphisms in the endothelial protein C receptor (EPCR) gene on the risk of venous thromboembolism in carriers of factor V Leiden.. Thromb Haemost 2005; 94: 389-394.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Miñano A, Ordóñez A, España F. et al. AB0 blood group and risk of venous or arterial thrombosis in carriers of factor V Leiden or prothrombin G20210A polymorphisms.. Haematologica 2008; 93: 729-734.
    CrossrefPubMedGoogle Scholar
  • 18 Smith NL, Hindorff LA, Heckbert SR. et al. Association of genetic variations with nonfatal venous thrombosis in postmenopausal women.. J Am Med Assoc 2007; 297: 489-498.
    CrossrefPubMedGoogle Scholar
  • 19 Bezemer ID, Bare LA, Doggen CJ. et al. Gene variants associated with deep vein thrombosis.. Jama 2008; 299: 1306-1314.
    CrossrefPubMedGoogle Scholar
  • 20 Germain M, Saut N, Greliche N. et al. Genetics of venous thrombosis: insights from a new genome wide association study.. PLoS One 2011; 6: e25581
    CrossrefPubMedGoogle Scholar
  • 21 Plomin R, Haworth CM, Davis OS. Common disorders are quantitative traits.. Nat Rev Genet 2009; 10: 872-878.
    CrossrefPubMedGoogle Scholar
  • 22 Wichers IM, Tanck MW, Meijers JC. et al. Assessment of coagulation and fibri-nolysis in families with unexplained thrombophilia.. Thromb Haemost 2009; 101: 465-470.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Segers O, van Oerle R, ten Cate H. et al. Thrombin generation as an intermediate phenotype for venous thrombosis.. Thromb Haemost 2010; 103: 114-122.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Brandts A, van Hylckama Vlieg A, Rosing J. et al. The risk of venous thrombosis associated with a high endogenous thrombin potential in the absence and presence of activated protein C.. J Thromb Haemost 2007; 5: 416-418.
    CrossrefPubMedGoogle Scholar
  • 25 Dargaud Y, Trzeciak MC, Bordet JC. et al. Use of calibrated automated thrombi-nography +/- thrombomodulin to recognise the prothrombotic phenotype.. Thromb Haemost 2006; 96: 562-567.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 van Hylckama Vlieg A, Christiansen SC, Luddington R. et al. Elevated endogenous thrombin potential is associated with an increased risk of a first deep venous thrombosis but not with the risk of recurrence.. Br J Haematol 2007; 138: 769-774.
    CrossrefPubMedGoogle Scholar
  • 27 Tripodi A, Martinelli I, Chantarangkul V. et al. The endogenous thrombin potential and the risk of venous thromboembolism.. Thromb Res 2007; 121: 353-359.
    CrossrefPubMedGoogle Scholar
  • 28 ten Cate-Hoek AJ, Dielis AW, Spronk HM. et al. Thrombin generation in patients after acute deep-vein thrombosis.. Thromb Haemost 2008; 100: 240-245.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Hron G, Kollars M, Binder BR. et al. Identification of patients at low risk for recurrent venous thromboembolism by measuring thrombin generation.. J Am Med Assoc 2006; 296: 397-402.
    CrossrefPubMedGoogle Scholar
  • 30 Eichinger S, Hron G, Kollars M. et al. Prediction of recurrent venous throm-boembolism by endogenous thrombin potential and D-dimer.. Clin Chem 2008; 54: 2042-2048.
    CrossrefPubMedGoogle Scholar
  • 31 Besser M, Baglin C, Luddington R. et al. High rate of unprovoked recurrent venous thrombosis is associated with a high thrombin generating potential in a prospective cohort study.. J Thromb Haemost 2008; 6: 1720-1725.
    CrossrefPubMedGoogle Scholar
  • 32 Tripodi A, Legnani C, Chantarangkul V. et al. High thrombin generation measured in the presence of thrombomodulin is associated with an increased risk of recurrent venous thromboembolism.. J Thromb Haemost 2008; 6: 1327-1333.
    CrossrefPubMedGoogle Scholar
  • 33 Lutsey PL, Folsom AR, Heckbert SR. et al. Peak thrombin generation and subsequent venous thromboembolism: the Longitudinal Investigation of Throm-boembolism Etiology (LITE).. J Thromb Haemost 2009; 7: 1639-1648.
    CrossrefPubMedGoogle Scholar
  • 34 Hemker HC, Giesen P, AlDieri R. et al. The calibrated automated thrombogram (CAT): a universal routine test for hyper- and hypocoagulability.. Pathophysiol Haemost Thromb 2002; 32: 249-253.
    CrossrefPubMedGoogle Scholar
  • 35 Castoldi E, Maurissen LF, Tormene D. et al. Similar hypercoagulable state and thrombosis risk in type I and type III protein S-deficient individuals from families with mixed type I/III protein S deficiency.. Haematologica 2010; 95: 1563-1571.
    CrossrefPubMedGoogle Scholar
  • 36 Dielis AW, Castoldi E, Spronk HM. et al. Coagulation factors and the protein C system as determinants of thrombin generation in a normal population.. J Thromb Haemost 2008; 6: 125-131.
    PubMedGoogle Scholar
  • 37 Stephens M, Smith NJ, Donnelly P. A new statistical method for haplotype reconstruction from population data.. Am J Hum Genet 2001; 68: 978-989.
    CrossrefPubMedGoogle Scholar
  • 38 Poort SR, Rosendaal FR, Reitsma PH. et al. A common genetic variation in the 3’-untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increase in venous thrombosis.. Blood 1996; 88: 3698-3703.
    PubMedGoogle Scholar
  • 39 Ceelie H, Bertina RM, van Hylckama Vlieg A. et al. Polymorphisms in the pro-thrombin gene and their association with plasma prothrombin levels.. Thromb Haemost 2001; 85: 1066-1070.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 40 Castoldi E, Simioni P, Tormene D. et al. Differential effects of high prothrombin levels on thrombin generation depending on the cause of the hyperprothrom-binemia.. J Thromb Haemost 2007; 5: 971-979.
    CrossrefPubMedGoogle Scholar
  • 41 Martinelli I, Battaglioli T, Tosetto A. et al. Prothrombin A19911G polymorphism and the risk of venous thromboembolism.. J Thromb Haemost 2006; 4: 2582-2586.
    CrossrefPubMedGoogle Scholar
  • 42 Chinthammitr Y, Vos HL, Rosendaal FR. et al. The association of prothrombin A19911G polymorphism with plasma prothrombin activity and venous thrombosis: results of the MEGA study, a large population-based case-control study.. J Thromb Haemost 2006; 4: 2587-2592.
    CrossrefPubMedGoogle Scholar
  • 43 Antón AI, Teruel R, Corral J. et al. Functional consequences of the prothrom-botic SERPINC1 rs2227589 polymorphism on antithrombin levels. Haemato- logica 2009; 94: 589-592.
    PubMedGoogle Scholar
  • 44 Bertina RM, Ploos van Amstel HK, van Wijngaarden A. et al. Heerlen polymorphism of protein S, an immunologic polymorphism due to dimorphism of residue 460. Blood 1990; 76: 538-548.
    PubMedGoogle Scholar
  • 45 Moatti D, Seknadji P, Galand C. et al. Polymorphisms of the tissue factor pathway inhibitor (TFPI) gene in patients with acute coronary syndromes and in healthy subjects : impact of the V264M substitution on plasma levels of TFPI. Arterioscler Thromb Vasc Biol 1999; 19: 862-869.
    CrossrefPubMedGoogle Scholar
  • 46 Giri TK, Yamazaki T, Sala N. et al. Deficient APC-cofactor activity of protein S Heerlen in degradation of factor Va Leiden: a possible mechanism of synergism between thrombophilic risk factors. Blood 2000; 96: 523-531.
    PubMedGoogle Scholar
  • 47 Arnaud E, Moatti D, Emmerich J. et al. No link between the TFPI V264M mutation and venous thromboembolic disease. Thromb Haemost 1999; 82: 159-160.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 48 Amini-Nekoo A, Futers TS, Moia M. et al. Analysis of the tissue factor pathway inhibitor gene and antigen levels in relation to venous thrombosis. Br J Haema-tol 2001; 113: 537-543.
    CrossrefPubMedGoogle Scholar
  • 49 Van Dreden P, Grosley M, Cost H. Total and free levels of tissue factor pathway inhibitor: a risk factor in patients with factor V Leiden?. Blood Coagul Fibrinolysis 1999; 10: 115-116.
    CrossrefPubMedGoogle Scholar
  • 50 Rosing J, Hoekema L, Nicolaes GA. et al. Effects of protein S and factor Xa on peptide bond cleavages during inactivation of factor Va and factor VaR506Q by activated protein C. J Biol Chem 1995; 270: 27852-27858.
    CrossrefPubMedGoogle Scholar