Download PDF
Thromb Haemost 2015; 113(04): 870-880
DOI: 10.1160/TH14-06-0519
Stroke, Systemic or Venous Thromboembolism
Schattauer GmbH

Defective thrombus formation in mice lacking endogenous factor VII activating protease (FSAP)

Saravanan Subramaniam
1   Institute of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Ina Thielmann
2   Rudolf Virchow Center, University of Würzburg, Germany
,
Martina Morowski
2   Rudolf Virchow Center, University of Würzburg, Germany
,
Ingo Pragst
3   CSL Behring GmbH, Marburg, Germany
,
Per Morten Sandset
4   University of Oslo and Oslo University Hospital, Oslo, Norway
,
Bernhard Nieswandt
2   Rudolf Virchow Center, University of Würzburg, Germany
,
Michael Etscheid
5   Paul Ehrlich Institute, Langen, Germany
,
Sandip M. Kanse
1   Institute of Biochemistry, Justus-Liebig-University, Giessen, Germany
4   University of Oslo and Oslo University Hospital, Oslo, Norway
› Author Affiliations Financial Support: Funding from the Deutsche Forschungsgemeinschaft (DFG), Behring Roentgen Stiftung (BRS) and Helse Sør-Øst to SMK is acknowledged.
Further Information

Publication History

Received: 16 June 2014

Accepted after major revision: 20 October 2014

Publication Date:
24 November 2017 (online)

Also available at
    Permissions and Reprints

Summary

Factor VII (FVII) activating protease (FSAP) is a circulating protease with a putative function in blood coagulation and fibrinolysis. Genetic epidemiological studies have implied a role for FSAP in carotid stenosis, stroke and thrombosis. To date, no in vivo evidence is available to support these claims. We have, for the first time, used FSAP-/- mice to define its role in thrombosis and haemostasis in vivo and to characterise the molecular mechanisms involved. FeCl3–induced arterial thrombosis in carotid and mesenteric artery revealed that the occlusion time was significantly increased in FSAP-/- mice (p< 0.01) and that some FSAP-/- mice did not occlude at all. FSAP-/- mice were protected from lethal pulmonary thromboembolism induced by collagen/ epinephrine infusion (p< 0.01). Although no spontaneous bleeding was evident, in the tail bleeding assay a re-bleeding pattern was observed in FSAP-/- mice. To explain these observations at a mechanistic level we then determined how haemostasis factors and putative FSAP substrates were altered in FSAP-/- mice. Tissue factor pathway inhibitor (TFPI) levels were higher in FSAP-/- mice compared to WT mice whereas FVIIa levels were unchanged. Other coagulation factors as well as markers of platelet activation and function revealed no significant differences between WT and FSAP-/- mice. This phenotype of FSAP-/- mice could be reversed by application of exogenous FSAP. In conclusion, a lack of endogenous FSAP impaired the formation of stable, occlusive thrombi in mice. The underlying in vivo effect of FSAP is more likely to be related to the modulation of TFPI rather than FVIIa.

Keywords

Coagulation - fibrinolysis - thrombosis - haemostasis - FSAP - HABP2

 

  • References

  • 1 Romisch J. Factor VII activating protease (FSAP): a novel protease in hemostasis. Biol Chem 2002; 383: 1119-1124.
    PubMedGoogle Scholar
  • 2 Kanse SM, Parahuleva M, Muhl L. et al. Factor VII-activating protease (FSAP): vascular functions and role in atherosclerosis. Thromb Haemost 2008; 99: 286-289.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Etscheid M, Kress J, Seitz R. et al. The hyaluronic acid-binding protease: a novel vascular and inflammatory mediator?. Int Immunopharmacol 2008; 08: 166-170.
    CrossrefPubMedGoogle Scholar
  • 4 Stephan F, Hazelzet JA, Bulder I. et al. Activation of factor VII-activating protease in human inflammation: a sensor for cell death. Crit Care 2011; 15: R110.
    CrossrefPubMedGoogle Scholar
  • 5 Zeerleder S, Zwart B, te Velthuis H. et al. Nucleosome-releasing factor: a new role for factor VII-activating protease (FSAP). FASEB J 2008; 22: 4077-4084.
    CrossrefPubMedGoogle Scholar
  • 6 Yamamichi S, Fujiwara Y, Kikuchi T. et al. Extracellular histone induces plasma hyaluronan-binding protein (factor VII activating protease) activation in vivo. Biochem Biophys Res Commun 2011; 409: 483-488.
    CrossrefPubMedGoogle Scholar
  • 7 Kanse SM, Gallenmueller A, Zeerleder S. et al. Factor VII-activating protease is activated in multiple trauma patients and generates anaphylatoxin C5a. J Immunol 2012; 188: 2858-2865.
    CrossrefPubMedGoogle Scholar
  • 8 Etscheid M, Muhl L, Pons D. et al. The Marburg I polymorphism of factor VII activating protease is associated with low proteolytic and low pro-coagulant activity. Thromb Res 2012; 130: 935-941.
    CrossrefPubMedGoogle Scholar
  • 9 Stavenuiter F, Dienava-Verdoold I, Boon-Spijker MG. et al. Factor seven activating protease (FSAP): does it activate factor VII?. J Thromb Haemost 2012; 10: 859-866.
    CrossrefPubMedGoogle Scholar
  • 10 Kanse SM, Declerck PJ, Ruf W. et al. Factor VII-activating protease promotes the proteolysis and inhibition of tissue factor pathway inhibitor. Arterioscl Thromb Vasc Biol 2012; 32: 427-433.
    CrossrefPubMedGoogle Scholar
  • 11 Stephan F, Dienava I, Bulder I. et al. Tissue factor pathway inhibitor is an inhibitor of Factor VII-activating protease. J Thromb Haemost 2012; 10: 7.
    PubMedGoogle Scholar
  • 12 Roemisch J, Feussner A, Nerlich C. et al. The frequent Marburg I polymorphism impairs the pro-urokinase activating potency of the factor VII activating protease (FSAP). Blood Coagul Fibrinol 2002; 13: 433-441.
    CrossrefPubMedGoogle Scholar
  • 13 Sedding D, Daniel JM, Muhl L. et al. The G534E polymorphism of the gene encoding the factor VII-activating protease is associated with cardiovascular risk due to increased neointima formation. J Exp Med 2006; 203: 2801-2807.
    CrossrefPubMedGoogle Scholar
  • 14 Willeit J, Kiechl S, Weimer T. et al. Marburg I polymorphism of factor VII--activating protease: a prominent risk predictor of carotid stenosis. Circulation 2003; 107: 667-670.
    CrossrefPubMedGoogle Scholar
  • 15 Wasmuth HE, Tag CG, Van de Leur E. et al. The Marburg I variant (G534E) of the factor VII-activating protease determines liver fibrosis in hepatitis C infection by reduced proteolysis of platelet-derived growth factor BB. Hepatology 2009; 49: 775-780.
    CrossrefPubMedGoogle Scholar
  • 16 Trompet S, Pons D, Kanse SM. et al. Factor VII Activating Protease Polymorphism (G534E) Is Associated with Increased Risk for Stroke and Mortality. Stroke Res Treatm 2011; 2011: 424759.
    PubMedGoogle Scholar
  • 17 Hoppe B, Tolou F, Radtke H. et al. Marburg I polymorphism of factor VII-activating protease is associated with idiopathic venous thromboembolism. Blood 2005; 105: 1549-1551.
    CrossrefPubMedGoogle Scholar
  • 18 Ahmad-Nejad P, Dempfle CE, Weiss C. et al. The G534E-polymorphism of the gene encoding the factor VII-activating protease is a risk factor for venous thrombosis and recurrent events. Thromb Res 2012; 130: 441-444.
    CrossrefPubMedGoogle Scholar
  • 19 van Minkelen R, de Visser MC, Vos HL. et al. The Marburg I polymorphism of factor VII-activating protease is not associated with venous thrombosis. Blood 2005; 105: 4898.
    CrossrefPubMedGoogle Scholar
  • 20 Gulesserian T, Hron G, Endler G. et al. Marburg I polymorphism of factor VIIactivating protease and risk of recurrent venous thromboembolism. Thromb Haemost 2006; 95: 65-67.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Franchi F, Martinelli I, Biguzzi E. et al. Marburg I polymorphism of factor VIIactivating protease and risk of venous thromboembolism. Blood 2006; 107: 1731.
    CrossrefPubMedGoogle Scholar
  • 22 Pecheniuk NM, Elias DJ, Xu X. et al. Failure to validate association of gene polymorphisms in EPCR, PAR-1, FSAP and protein S Tokushima with venous thromboembolism among Californians of European ancestry. Thromb Haemost 2008; 99: 453-455.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Weisbach V, Ruppel R, Eckstein R. The Marburg I polymorphism of factor VIIactivating protease and the risk of venous thromboembolism. Thromb Haemost 2007; 97: 870-872.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Borkham-Kamphorst E, Zimmermann HW, Gassler N. et al. Factor VII activating protease (FSAP) exerts anti-inflammatory and anti-fibrotic effects in liver fibrosis in mice and men. J Hepatol 2013; 58: 104-111.
    CrossrefPubMedGoogle Scholar
  • 25 Maroney SA, Ferrel JP, Pan S. et al. Temporal expression of alternatively spliced forms of tissue factor pathway inhibitor in mice. J Thromb Haemost 2009; 07: 1106-1113.
    CrossrefPubMedGoogle Scholar
  • 26 Girard TJ, Tuley E, Broze Jr. GJ. TFPIbeta is the GPI-anchored TFPI isoform on human endothelial cells and placental microsomes. Blood 2012; 119: 1256-1262.
    CrossrefPubMedGoogle Scholar
  • 27 Broze Jr. GJ, Girard TJ. Tissue factor pathway inhibitor: structure-function. Front Biosci 2012; 17: 262-280.
    CrossrefPubMedGoogle Scholar
  • 28 Massberg S, Grahl L, von Bruehl ML. et al. Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases. Nature Med 2010; 16: 887-896.
    CrossrefPubMedGoogle Scholar
  • 29 Vincent LM, Tran S, Livaja R. et al. Coagulation factor V(A2440G) causes east Texas bleeding disorder via TFPIalpha. J Clin Invest 2013; 123: 3777-3787.
    CrossrefPubMedGoogle Scholar
  • 30 Wood JP, Bunce MW, Maroney SA. et al. Tissue factor pathway inhibitor-alpha inhibits prothrombinase during the initiation of blood coagulation. Proc Natl Acad Sci USA 2013; 110: 17838-17843.
    CrossrefPubMedGoogle Scholar
  • 31 Westrick RJ, Winn ME, Eitzman DT. Murine models of vascular thrombosis (Eitzman series). Arterioscl Thromb Vasc Biol 2007; 27: 2079-2093.
    CrossrefPubMedGoogle Scholar