Download PDF
Thromb Haemost 2002; 88(03): 462-472
DOI: 10.1055/s-0037-1613239
Review Article
Schattauer GmbH

Mice with a Severe Deficiency of the Endothelial Protein C Receptor Gene Develop, Survive, and Reproduce Normally, and Do not Present with Enhanced Arterial Thrombosis after Challenge[*]

Francis J. Castellino
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
Zhong Liang
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
S. Patrick Volkir
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
Erijka Haalboom
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
J. Andrew Martin
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
Mayra J. Sandoval-Cooper
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
,
Elliot D. Rosen
1   W. M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA
› Author Affiliations
Further Information

Publication History

Received 14 March 2002

Accepted after revision 30 April 2002

Publication Date:
27 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

The endothelial cell Protein C receptor (EPCR) functions to enhance activation of anticoagulant Protein C (PC) by the thrombin/ thrombomodulin (Tm) complex on the surface of the endothelium. This overall system functions in anticoagulation, profibrinolytic, and antiinflammatory responses. Mice with a severe targeted deficiency of this receptor have been generated by integration of exogenous DNA elements into the 5’-untranslated region of the EPCR gene. Despite the retention of the entire endogenous EPCR coding sequence in the altered EPCR gene locus, only very low EPCR message contents were detected in mice by quantitative RT-PCR during embryogenesis and up to at least early adulthood. Immunohistochemical analysis of various regions of the arterial tree of mice up to 4 months of age, employing an anti-murine EPCR antibody, confirmed that undetectable levels of this protein were present in arterial regions during these periods. Despite this, these mice are not more prone to arterial thrombosis after challenge in a FeCl3 carotid artery thrombosis model. Small amounts (<10% of wild-type) of this protein were found in other tissues. Matings of mice homozygous for this deficiency led to normal births and survival of the offspring, in contrast to results by others demonstrating early embryonic lethality of a total EPCR deficiency. These data further show that minimal levels of EPCR are able to support male and female virility, as well as embryonic development, birth, and survival to adulthood.

Keywords

EPCR - gene deficiency - embryogenesis - anticoagulation - reproduction

* This work was supported by grants HL-19982 (to F. J. C.), HL-65241 (to E. D. R./F. J. C.) and HL60081 (to E. D. R.) from the National Institutes of Health, and the Kleiderer-Pezold endowed chair (to F. J. C.).


 

  • References

  • 1 Castellino FJ. Human protein C and activated Protein C. Components of the human anticoagulation system. Trends Cardiovasc Med 1995; 05: 55-62.
    CrossrefPubMedGoogle Scholar
  • 2 Kisiel W, Davie EW. Protein C. Meth Enzymol 1981; 80: 320-32.
    PubMedGoogle Scholar
  • 3 Vehar GA, Davie EW. Preparation and properties of bovine factor VIII (antihemophilic factor). Biochemistry 1980; 19: 401-10.
    CrossrefPubMedGoogle Scholar
  • 4 Taylor FB, Lockhart MS. A new function for activated Protein C: Activated protein C prevents inhibition of plasminogen activators by releasate from mononuclear leukocytes-platelet suspensions stimulated by phorbol diester. Thromb Res 1985; 37: 639-49.
    CrossrefPubMedGoogle Scholar
  • 5 Bajzar L, Kalafatis M, Simioni P, Tracy PB. An antifibrinolytic mechanism describing the prothrombotic effect associated with factor V-Leiden. J Biol Chem 1996; 271: 22949-52.
    CrossrefPubMedGoogle Scholar
  • 6 Castellino FJ. Gene targeting in hemostasis: Protein C. Front. Biosci 2001; 06: D807-19.
    PubMedGoogle Scholar
  • 7 Jalbert LR, Rosen ED, Moons L, Chan JCY, Carmeliet P, Collen D, Castellino FJ. Inactivation of the gene for anticoagulant Protein C causes lethal perinatal consumptive coagulopathy in mice. J Clin Invest 1998; 102: 1481-8.
    CrossrefPubMedGoogle Scholar
  • 8 Fukudome K, Esmon CT. Identification, cloning, and regulation of a novel endothelial cell Protein C/activated Protein C receptor. J Biol Chem 1994; 269: 26486-91.
    PubMedGoogle Scholar
  • 9 Stearns-Kurosawa DJ, Kurosawa S, Mollica JS, Ferrell GL, Esmon CT. The endothelial cell protein C receptor augments protein C activation by the thrombin-thrombomodulin complex. Proc Natl Acad Sci USA 1996; 93: 10212-6.
    CrossrefPubMedGoogle Scholar
  • 10 Fukudome K, Ye XF, Tsuneyoshi N, Tokunaga O, Sugawara K, Mizokami H, Kimoto M. Activation mechanism of anticoagulant Protein C in large blood vessels involving the endothelial cell protein C receptor. J Exp Med 1998; 187: 1029-35.
    CrossrefPubMedGoogle Scholar
  • 11 Hayashi T, Nakamura H, Okada A, Takebayashi S, Wakita T, Yuasa H, Okumura K, Suzuki K. Organization and chromosomal localization of the human endothelial Protein C receptor gene. Gene 1999; 238: 367-73.
    CrossrefPubMedGoogle Scholar
  • 12 Liang Z, Rosen ED, Castellino FJ. Nucleotide structure and characterization of the murine gene encoding the endothelial cell Protein C receptor. Thromb Haemost 1999; 81: 585-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Xu J, Qu DF, Esmon NL, Esmon CT. Metalloproteolytic release of endothelial cell Protein C receptor. J Biol Chem 2000; 275: 6038-44.
    CrossrefPubMedGoogle Scholar
  • 14 Liaw PCY, Neuenschwander PF, Smirnov MD, Esmon CT. Mechanisms by which soluble endothelial cell Protein C receptor modulates Protein C and activated Protein C function. J Biol Chem 2000; 275: 5447-52.
    CrossrefPubMedGoogle Scholar
  • 15 Taylor FB, Chang A, Esmon CT, D’Angelo A, Vigano-D’Angelo S, Blick KE. Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J Clin Invest 1987; 79: 918-25.
    CrossrefPubMedGoogle Scholar
  • 16 Kurosawa S, Stearns-Kurosawa DJ, Carson CW, DAngelo A, DellaValle P, Esmon CT. C4b-binding protein exacerbates the host response to Escherichia coli. Blood 1988; 91: 725-7.
    PubMedGoogle Scholar
  • 17 Taylor FB, Stearns-Kurosawa DJ, Kurosawa S, Ferrell G, Chang ACK, Laszik Z, Kosanke S, Peer G, Esmon CT. The endothelial cell Protein C receptor aids in host defense against Escherichia coli sepsis. Blood 2000; 95: 1680-6.
    PubMedGoogle Scholar
  • 18 Kurosawa S, Esmon CT, Stearns-Kurosawa DJ. Plasma levels of endothelial cell Protein C receptor are elevated in patients with sepsis and systemic lupus erythematosus: Lack of correlation with thrombomodulin suggests involvement of different pathological processes. J Immunol 1998; 165: 4697-703.
    PubMedGoogle Scholar
  • 19 Gu JM, Ferrell G, Esmon CT. The soluble endothelial protein C receptor binds to activated neutrophils: Involvement of proteinase-3 and CD11b/CD18. Blood 2001; 96: 814a.
    PubMedGoogle Scholar
  • 20 Rosen E, Chan JC, Idusogie E, Clotman F, Vlasuk G, Luther T, Jalbert L, Albrecht S, Zhong L, Lissens A, Schoonjans L, Moons L, Collen D, Castellino FJ, Carmeliet P. Mice lacking factor VII develop normally but suffer fatal perinatal bleeding. Nature 1997; 390: 290-4.
    CrossrefPubMedGoogle Scholar
  • 21 Dewerchin M, Liang Z, Moons L, Carmeliet P, Castellino FJ, Collen D, Rosen ED. Blood coagulation factor X deficiency causes partial embryonic lethality and fatal neonatal bleeding in mice. Thromb Haemost 2000; 83: 185-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Ploplis VA, Wilberding J, McLennan L, Liang Z, Cornelissen I, DeFord ME, Rosen ED, Castellino FJ. A total fibrinogen deficiency is compatible with the development of pulmonary fibrosis in mice. Am J Pathol 2000; 157: 703-8.
    CrossrefPubMedGoogle Scholar
  • 23 Fay W, Parker AC, Ansari MN, Zheng X, Ginsburg D. Vitronectin inhibits the thrombotic response to arterial injury in mice. Blood 1999; 93: 1825-30.
    PubMedGoogle Scholar
  • 24 Rothbarth K, Dabaghian ARH, Stammer H, Werner D. One single mRNA encodes the centrosomal protein CCD41 and the endothelial cell Protein C receptor (EPCR). FEBS Lett 1999; 458: 77-80.
    CrossrefPubMedGoogle Scholar
  • 25 Parry GC, Erlich JH, Carmeliet P, Luther T, Mackman N. Low levels of tissue factor are compatible with development and hemostasis in mice. J Clin Invest 1998; 101: 560-9.
    CrossrefPubMedGoogle Scholar
  • 26 Merati G, Biguzzi E, Oganesyan N, Fetiveau R, Qu D, Bucciarelli P, Stearns-Kurosawa DJ, Mannucci PM, Esmon CT, Fainoni EM. A 23bp insertion in the endothelial Protein C receptor (EPCR) gene in patients with myocardial infarction and deep vein thrombosis. Thromb Heamost 1999; Supp. August 1999: 507.
    PubMedGoogle Scholar
  • 27 Van de Water NS, French JK, McDowell J, Browett PJ. The endothelial Protein C receptor (EPCR) 23bp insert in patients with myocardial infarction. Thromb Haemost 2001; 85: 749-51.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 von Depka M, Czwalinna A, Eisert R, Wermes C, Scharrer I, Ganser A, Ehrenforth S. Prevalence of a 23bp insertion in exon 3 of the endothelial cell Protein C receptor gene in venous thrombophilia. Thromb. Haemost 2001; 86: 1360-2.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Isermann B, Hendrickson SB, Hutley K, Wing M, Weiler H. Tissue-restricted expression of thrombomodulin in the placenta rescues thrombomodulindeficient mice from early lethality and reveals a secondary developmental block. Development 2001; 128: 827-38.
    PubMedGoogle Scholar
  • 30 Shu F, Kobayashi H, Fukudome K, Tsuneyoshi N, Kimoto M, Terao T. Activated Protein C suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner. FEBS Lett 2001; 47: 208-12.
    PubMedGoogle Scholar
  • 31 Murakami K, Okajima K, Uchiba M, Johno M, Nakagaki T, Okabe H, Takatsuki K. Activated Protein C prevents LPS-induced pulmonary vascular injury by inhibiting cytokine production. Am J Physiol 1997; 16: L197-02.
    PubMedGoogle Scholar
  • 32 Hancock WW, Grey ST, Hau L, Akalin E, Orthner C, Sayegh MH, Salem HH. Binding of activated Protein C to a specific receptor on human mononuclear phagocytes inhibits intracellular calcium signaling and monocyte-dependent proliferative responses. Transplantation 1995; 60: 1525-32.
    CrossrefPubMedGoogle Scholar