Blood Coagulation Factor X Deficiency Causes Partial Embryonic Lethality and Fatal Neonatal Bleeding in Mice

Summary

Mice with a total deficiency in blood coagulation Factor X (FX) were generated by targeted replacement of an 18-kb fragment of the FX gene, comprising all exons encoding the mature FX protein, with a neo^r cassette. The genotype distribution among the offspring from heterozygous breeding pairs suggested that FX deficiency resulted in partial embryonic lethality, with approximately one-third of the FX ^−/− embryos dying around embryonic day (E) 11.5-12.5. Two of 44 non-resorbed FX ^−/− embryos analyzed at these stages showed signs of massive bleeding, one of which into the brain ventricles, but no histological defects in the vasculature of these embryos or their yolk sac were observed. The remainder of the FX ^−/− embryos appeared normal and survived to term, but the majority of neonates (90%) died within 5 days, most frequently from intraabdominal bleeding. The remaining FX ^−/− animals succumbed between postnatal day (P)5 and P20 with intraabdominal, subcutaneous, or intracranial bleeding or a combination thereof. The lethal phenotype of the FX ^−/− mice illustrates the importance of FX function in embryonic and postnatal survival and demonstrates that these mice serve as effective models of the bleeding disorders observed in severe FX deficiency in humans.

Keywords

Factor X - embryonic development - neonatal bleeding - gene deletion - coagulation

• References

• 1 McMullen BA, Fujikawa K, Kisiel W, Sasagawa T, Howald WN, Kwa EW, Weinstein B. Complete amino acid sequence of the light chain of human blood coagulation factor X: Evidence for identification of residue 63 as β-hydroxyaspartic acid. Biochemistry 1983; 22: 2875-84.
• 2 Leytus SP, Chung DW, Kisiel W, Kurachi K, Davie EW. Characterization of a cDNA coding for human factor X. Proc Natl Acad Sci USA 1984; 81: 3699-702.
• 3 Broze GJ, Girard TJ, Novotny WF. The lipoprotein-associated coagulation inhibitor. Prog Hemost Thromb 1991; 10: 243-68.
• 4 Broze Jr GJ. Tissue factor pathway inhibitor and the revised theory of coagulation. Annu Rev Med 1995; 46: 103-12.
• 5 Fujikawa K, Coan MH, Legaz ME, Davie EW. The mechanism of activation of bovine factor X (Stuart factor) by intrinsic and extrinsic pathways. Biochemistry 1974; 13: 5290-9.
• 6 Davie EW. Biochemical and molecular aspects of the coagulation cascade. Thromb Haemost 1995; 74: 1-6.
• 7 Gasic GP, Arenas CP, Gasic TB, Gasic GJ. Coagulation factors X, Xa, and protein S as potent mitogens of cultured aortic smooth muscle cells. Proc Natl Acad Sci USA 1992; 89: 2317-20.
• 8 Ko FN, Yang YC, Huang SC, Ou JT. Coagulation factor Xa stimulates plateletderived growth factor release and mitogenesis in cultured vascular smooth muscle cells of rat. J Clin Invest 1996; 98: 1493-501.
• 9 Ragosta M, Gimple LW, Gertz SD, Dunwiddie CT, Vlasuk GP, Haber HL, Powers ER, Roberts WC, Sarembock IJ. Specific factor Xa inhibition reduces restenosis after balloon angioplasty of atherosclerotic femoral arteries in rabbits. Circulation 1994; 89: 1261-71.
• 10 Herbert J, Bono F, Herault J, Avril C, Dol F, Mares A, Schaeffer P. Effector protease receptor 1 mediates the mitogenic activity of factor Xa for vascular smooth muscle cells in vitro and in vivo. J Clin Invest 1998; 101: 993-1000.
• 11 Cirino G, Cicala C, Bucci M, Sorrentino L, Ambrosini G. Factor Xa as an interface between coagulation and inflammation. Molecular mimicry of factor Xa association with effector cell protease receptor-1 induces acute inflammation in vivo. J Clin Invest 1997; 99: 2446-51.
• 12 Senden NH, Jeunhomme TM, Heemskerk JW, Wagenvoord R, van’t Veer C, Hemker HC, Buurman WA. Factor Xa induces cytokine production and expression of adhesion molecules by human umbilical vein endothelial cells. J Immunol 1998; 161: 4318-24.
• 13 Leytus SP, Foster DC, Kurachi K, Davie EW. Gene for human factor X: A blood coagulation factor whose gene organization is essentially identical with that of factor IX and protein C. Biochemistry 1986; 25: 5098-102.
• 14 Cooper A, Liang Z, Castellino FJ, Rosen ED. Cloning and Characterization of the Murine Coagulation Factor X Gene. Thromb Haemost. 2000 in press.
• 15 Liang Z, Cooper A, Rosen ED, Castellino FJ. Chromosomal arrangement of the murine coagulation factor VII and factor X genes. Thromb Haemost 1998; 80: 524-5.
• 16 Furie B. Vitamin K-dependent blood coagulation proteins: normal function and clinical disorders. In Blood: Principles and Practice of Hematology. Handin RI, Lux SE, Stossel TP. eds. Philadelphia: JB Lippincott; 1995: 1181-217.
• 17 Rosen E, Chan JCY, 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.
• 18 Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian HH. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia a. Nat Genet 1995; 10: 119-21.
• 19 Lin HF, Maeda N, Smithies O, Straight DL, Stafford DW. A coagulation factor IXdeficient mouse model for human hemophilia B. Blood 1997; 90: 3962-6.
• 20 Kundu RK, Sangiorgi F, Wu LY, Kurachi K, Anderson WF, Maxson R, Gordon EM. Targeted inactivation of the coagulation factor IX gene causes hemophilia B in mice. Blood 1998; 92: 168-74.
• 21 Cui JS, Oshea KS, Purkayastha A, Saunders TL, Ginsburg D. Fatal haemorrhage and incomplete block to embryogenesis in mice lacking coagulation factor V. Nature 1996; 384: 66-8.
• 22 Xue JC, Wu QY, Wesrfield LA, Tuley EA, Lu DS, Zhang Q, Shim K, Zheng XL, Sadler JE. Incomplete embryonic lethality and fatal neonatal hemorrhage caused by prothrombin deficiency in mice. Proc Natl Acad Sci USA 1998; 95: 7603-7.
• 23 Sun WY, Witte DP, Degen JL, Colbert MC, Burkart MC, Holmback K, Xiao Q, Bugge TH, Degen SJF. Prothrombin deficiency results in embryonic and neonatal lethality in mice. Proc Natl Acad Sci USA 1998; 95: 7597-602.
• 24 Connolly AJ, Ishihara H, Kahn ML, Farese RVJ, Coughlin SR. Role of the thrombin receptor in development and evidence for a second receptor. Nature 1996; 381: 516-9.
• 25 Huang Z-F, Higuchi D, Lasky N, Broze Jr GJ. Tissue factor pathway inhibitor gene disruption produces intrauterine lethality in mice. Blood 1997; 90: 944-51.
• 26 Carmeliet P, Mackman N, Moons L, Luther T, Gressens P, Van Vlaenderen I, De Munck H, Kasper M, Breier G, Evrard P, Muller M, Risau W, Edgington T, Collen D. Role of tissue factor in embryonic blood vessel development. Nature 1996; 383: 73-5.
• 27 Liang Z, Cooper A, DeFord ME, Carmeliet P, Collen D, Castellino FJ, Rosen ED. Cloning and characterization of a cDNA encoding murine coagulation factor X. Thromb Haemost 1998; 80: 87-91.
• 28 Mullen CA, Kilstrup M, Blaese RM. Transfer of the bacterial gene for cytosine deaminase to mammalian cells confers lethal sensitivity to 5-fluorocytosine: a negative selection system. Proc Natl Acad Sci USA 1992; 89: 33-7.
• 29 Jalbert LR, Rosen ESD, 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.
• 30 Carmeliet P, Kiekens L, Schoonjans L, Ream B, Van Nuffelen A, Prendergast G, Cole M, Bronson R, Collen D, Mulligan RC. Plasminogen activator inhibitor-1 genedeficient. I. Generation by homologous recombination and characterizationmice. J Clin Invest 1993; 92: 2746-55.
• 31 Chen S, Evans G. Use of polymerase chain reaction for screening transgenic mice. In: Methods in Molecular Biology. PCR Protocols: Current methods and applications. White BA. ed. Totowa; Humana Press: 1993. 15 75-80.
• 32 Carmeliet P, Ng YS, Nuyens D, Theilmeier G, Brusselmans K, Cornelissen I, Ehler E, Kakkar VV, Stalmans I, Mattot V, Perriard JC, Dewerchin M, Flameng W, Nagy A, Lupu F, Moons L, Collen D, D’Amore PA, Shima DT. Impaired myocardial angiogenesis and ischemic cardiomyopathy in mice lacking the vascular endothelial growth factor isoforms VEGF164 and VEGF188. Nat Med 1999; 05: 495-502.
• 33 Chan JCY, Carmeliet P, Moons L, Rosen ED, Huang Z-F, Broze GJ, Collen D, Castellino FJ. Factor VII deficiency rescues the intrauterine lethality in mice associated with a tissue factor pathway inhibitor deficit. J Clin Invest 1999; 103: 475-82.
• 34 Xue JC, Kalafatis M, Silveira JR, Kung C, Mann KG. Determination of the disulfide bridges in factor Va heavy chain. Biochemistry 1994; 33: 13109-16.
• 35 Suh TT, Holmback K, Jensen NJ, Daugherty CC, Small K, Simon DI, Potter S, Degen J. Resolution of spontaneous bleeding events but failure of pregnancy in fibrinogen-deficient mice. Genes Dev 1995; 09: 2020-30.
• 36 Shivdasani RA, Orkin SH. Erythropoiesis and globin gene expression in mice lacking the transcription factor NF-E2. Proc Natl Acad Sci USA 1995; 92: 8690-4.
• 37 Healy AM, Rayburn HB, Rosenberg RD, Weiler H. Absence of the blood-clotting regulator thrombomodulin causes embryonic lethality in mice before development of a functional cardiovascular system. Proc Natl Acad Sci USA 1995; 92: 850-4.
• 38 Gailani D, Lasky NM, Broze Jr GJ. A murine model of factor XI deficiency. Blood Coagul Fibrinolysis 1997; 08: 134-44.