Blood coagulation at the site of microvascular injury in healthy and coumadin-treated subjects heterogenous for factor V Leiden mutation

 

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Summary

There is little knowledge regarding the in-vivo course of coagulation reactions in subjects with factor (F)V Leiden (G1691A, R506Q). The aim of the current study was to evaluate the effect of FV Leiden on coagulant reactions triggered by vascular injury. At the site of microvascular injury, prothrombin activation and FVa formation and inactivation have been evaluated in 16 apparently healthy subjects and 16 patients on long-term anticoagulation with acenocoumarol, with eight heterogenous carriers of FV Leiden in the healthy and in the anticoagulated group. Thrombin formation, measured by levels of thrombin-antithrombin complexes (TAT) and thrombin B-chain, proceeded at similar rates in the bleeding-time blood in carriers and non-carriers of FV Leiden. The onset and rate of FV activation did not differ significantly among the four groups. When healthy carriers of FV Leiden were compared to healthy non-carriers, the onset of FVa inactivation by activated protein C (APC) was delayed by 60 to 90 seconds (p=0.01). Anticoagulated individuals also showed the same pattern of FV Leiden associated differences in the time of occurrence of the 30 kD FVa heavy-chain fragment (p=0.021). Our results indicate that when blood clotting is triggered by microvascular injury, the heterozygous expression of FV Leiden has no effect on thrombin generation in healthy or coumadintreated subjects. Inactivation of FVa by theAPC mechanism is attenuated significantly in carriers of FV Leiden but the magnitude of this effect is smaller than that observed in most purified systems.

• References

• 1 Kalafatis M, Mann KG. Factor V: a combination of Dr Jeckyll and Mr Hyde. Blood 2003; 101: 20-30.
  CrossrefPubMedGoogle Scholar
• 2 Nicolaes GAF, Dahlbäck B. Factor V and thrombotic disease. Description of a Janus-face protein. Arterioscler Thromb Vasc Biol 2002; 22: 530-538.
  CrossrefPubMedGoogle Scholar
• 3 Kalafatis M, Rand MD, Mann KG. The mechanism of inactivation of human factor V and human factor Va by activated protein C. J Biol Chem 1994; 269: 31869-31880.
  PubMedGoogle Scholar
• 4 Bertina RM, Koelman BP, Rosendaal FR. et al. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature 1994; 369: 64-67.
  CrossrefPubMedGoogle Scholar
• 5 Voorberg J, Roelse J, Koopman R. et al. Association of idiopathic venous thromboembolism with single point-mutation at Arg506 of factor V. Lancet 1994; 343: 1535-1536.
  CrossrefPubMedGoogle Scholar
• 6 Martinelli I. Risk factors in venous thromboembolism. Thromb Haemost 2001; 86: 395-403.
  Article in Thieme ConnectPubMedGoogle Scholar
• 7 Kalafatis M, Bertina RM, Rand MD. et al. Characterization of the molecular defect in factor V506Q. J Biol Chem 1995; 270: 4053-4057.
  CrossrefPubMedGoogle Scholar
• 8 Heeb MJ, Koijma Y, Greengard JS. et al. Activated protein C resistance: Molecular mechanisms based on studies using purified Gln506-factor V. Blood 1995; 85: 3405-3411.
  PubMedGoogle Scholar
• 9 Van ’t Veer C, Kalafatis M, Bertina RM. et al. Increased tissue factor-initiated prothrombin activation as a result of the Arg506→Gln mutation in factor V Leiden. J Biol Chem 1997; 272: 20721-20729.
  CrossrefPubMedGoogle Scholar
• 10 Camire RM, Kalafatis M, Simioni P. et al. Plateletderived factor V/Va Leiden cofactor activities are sustained on the surface of activated platelets despite the presence of activated protein C. Blood 1998; 91: 2818-2829.
  PubMedGoogle Scholar
• 11 Greengard JS, Eichinger S, Griffin JH. et al. Variability of thrombosis among homozygous siblings with resistance to activated protein C due to Arg→Gln mutation in the gene for factor V. N Engl J Med 1994; 331: 1559-1562.
  CrossrefPubMedGoogle Scholar
• 12 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
• 13 Nicolaes GAF, 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
• 14 Martinelli I, Bottasso B, Duca F. et al. Heightened thrombin generation in individuals with resistance to activated protein C. Thromb Haemost 1996; 75: 703-705.
  Article in Thieme ConnectPubMedGoogle Scholar
• 15 Zöller B, Holm J, Svensson P. et al. Elevated levels of prothrombin activation fragment 1+2 in plasma from patients with heterozygous Arg506 to Gln mutation in the factor V gene (APC-resistance) and/or inherited protein S deficiency. Thromb Haemost 1996; 75: 270-274.
  Article in Thieme ConnectPubMedGoogle Scholar
• 16 Nicolaes GAF, Thomassen MC, Tans G. et al. Effect of activated protein C on thrombin generation and on thrombin potential in plasma of normal and APC-resistant individuals. Blood Coagul Fibrinolysis 1997; 8: 28-38.
  CrossrefPubMedGoogle Scholar
• 17 Kyrle PA, Eichinger S, Pabinger I. et al. Prothrombin fragment F1+2 is not predictive for recurrent venous thromboembolism. Thromb Haemost 1997; 77: 829-833.
  Article in Thieme ConnectPubMedGoogle Scholar
• 18 Petaja J, Hakala L, Rasi V. et al. Circulating activated protein C in subjects with heterozygous Gln506-factor V. Haemostasis 1998; 28: 31-36.
  PubMedGoogle Scholar
• 19 Undas A, Brummel K, Musial J, Mann KG, Szczeklik A. Blood coagulation at the site of microvascular injury: effects of low-dose aspirin. Blood 2001; 98: 2423-2431.
  CrossrefPubMedGoogle Scholar
• 20 Weiss JH, Lages B. Evidence for tissue factor-dependent activation of the classic extrinsic coagulation mechanism in blood obtained from bleeding time wounds. Blood 1988; 71: 629-635.
  PubMedGoogle Scholar
• 21 Szczeklik A, Krzanowski M, Góra P. et al. Antiplatelet drugs and generation of thrombin in clotting blood. Blood 1992; 80: 2006-2011.
  PubMedGoogle Scholar
• 22 Eichinger S, Wolz M, Nieszpaur-Los M. et al. Effects of a low molecular weight heparin (Fragmin) and of unfractionated heparin on coagulation activation at the site of plug formation in vivo. Thromb Haemost 1994; 72: 831-835.
  Article in Thieme ConnectPubMedGoogle Scholar
• 23 Undas A, Brummel K, Musial J. et al. Simvastatin impairs blood coagulation by inhibiting activation of prothrombin, factor V, factor XIII, and by enhancing factor Va inactivation. Circulation 2001; 103: 2248-2253.
  CrossrefPubMedGoogle Scholar
• 24 Kalafatis M, Bernardi F, Simioni P. et al. Phenotype and genotype expression in pseudohomozygous factor VLeiden. The need for phenotype analysis. Arterioscler Thromb Vasc Biol 1999; 19: 336-342.
  CrossrefPubMedGoogle Scholar
• 25 Rand MD, Lock JB, van’t Veer C. et al. Blood clotting in minimally altered whole blood. Blood 1996; 88: 3432-3445.
  PubMedGoogle Scholar
• 26 Hockin MF, Kalafatis M, Shatos M. et al. Protein C activation and factor Va inactivation on human umbilical vein endothelial cells. Arterioscler Thromb Vasc Biol 1997; 17: 2765-2775.
  CrossrefPubMedGoogle Scholar
• 27 Brummel KE, Paradis SG, Butenas S. et al. Thrombin functions during tissue factor-induced blood coagulation. Blood 2002; 100: 148-152.
  CrossrefPubMedGoogle Scholar
• 28 Catto A, Carter A, Ireland H. et al. Factor V Leiden gene mutation and thrombin generation in relation to the development of acute stroke. Arterioscler Thromb Vasc Biol 1995; 15: 783-785.
  CrossrefPubMedGoogle Scholar
• 29 Juul K, Tybjaerg-Hansen A, Steffensen R. et al. Factor V Leiden: The Copenhagen City Heart Study and 2 meta-analyses. Blood 2002; 100: 3-10.
  CrossrefPubMedGoogle Scholar
• 30 Slungaard A, Fernandez JA, Griffin JH. et al. Platelet factor 4 enhances generation of activated protein C in vitro and vivo. Blood 2003; 102: 146-151.
  CrossrefPubMedGoogle Scholar
• 31 Tracy PB, Nesheim ME, Mann KG. Proteolytic alterations of factor Va bound to platelets. J Biol Chem 1983; 258: 662-669.
  PubMedGoogle Scholar
• 32 Esmon CT. Molecular mechanisms that control the protein C anticoagulant pathway. Thromb Haemost 1993; 70: 29-35.
  PubMedGoogle Scholar