Intravascular coagulation activation in a murine model of thrombomodulin deficiency: effects of lesion size, age, and hypoxia on fibrin deposition.

We consecutively inactivated both alleles of the thrombomodulin (TM) gene in murine embryonic stem (ES) cells and generated TM-deficient (TM-/-) chimeric mice. Quantitation of an ES-cell marker and protein C cofactor activity indicates that up to 50% of pulmonary endothelial cells are ES-cell derived and therefore TM deficient. Infusions of 125I-fibrinogen into mice show a significant increase (fourfold, P <.005) in radiolabeled cross-linked fibrin in TM-/- chimeric mouse lung as compared with wild-type mice. However, only chimeric mice that exhibit at least a 30% reduction in protein C cofactor activity and are at least 15 months old display this phenotype. Immunocytochemical localization of TM in chimeras shows a mosaic pattern of expression in both large and small blood vessels. Colocalization of cross-linked fibrin and neo (used to replace TM) reveals that fibrin is deposited in TM-/- regions. However, the fibrin deposits were largely restricted to pulmonary vessels with a lumenal area greater than 100 micrometer2. The hypercoagulable phenotype can be induced in younger chimeric mice by exposure to hypoxia, which causes a fivefold increase in beta-fibrin levels in lung. Our findings show that TM chimerism results in spontaneous, intravascular fibrin deposition that is dependent on age and the magnitude of the TM deficiency.

[1]  H. Rayburn,et al.  A targeted point mutation in thrombomodulin generates viable mice with a prethrombotic state. , 1998, The Journal of clinical investigation.

[2]  W. Aird,et al.  Vascular Bed–specific Expression of an Endothelial Cell Gene Is Programmed by the Tissue Microenvironment , 1997, The Journal of cell biology.

[3]  Y. S. Zhou,et al.  Monocytes and tissue factor promote thrombosis in a murine model of oxygen deprivation. , 1997, The Journal of clinical investigation.

[4]  W. Aird,et al.  Developmentally regulated gene expression of thrombomodulin in postimplantation mouse embryos. , 1996, Development.

[5]  E J Boland,et al.  Age-specific regulation of clotting factor IX gene expression in normal and transgenic mice. , 1995, Blood.

[6]  P M Mannucci,et al.  Hypercoagulability in centenarians: the paradox of successful aging. , 1995, Blood.

[7]  D. Pinsky,et al.  Induction of interleukin 6 (IL-6) by hypoxia in vascular cells. Central role of the binding site for nuclear factor-IL-6 , 1995, The Journal of Biological Chemistry.

[8]  R. Rosenberg,et al.  The absence of the blood clotting regulator thrombomodulin causes embryonic lethality in mice before development of a functional cardiovascular system. , 1995, Thrombosis and haemostasis.

[9]  R. Marlar,et al.  THE FIRST MUTATION IDENTIFIED IN THE THROMBOMODULIN GENE IN A 45-YEAR-OLD MAN PRESENTING WITH THROMBOEMBOLIC DISEASE , 1995 .

[10]  P. Carmeliet,et al.  Physiological consequences of loss of plasminogen activator gene function in mice , 1994, Nature.

[11]  K. Kario,et al.  Lipid-related hemostatic abnormalities in the elderly: imbalance between coagulation and fibrinolysis , 1993 .

[12]  J. Griffin,et al.  Normalization of markers of coagulation activation with a purified protein C concentrate in adults with homozygous protein C deficiency , 1993 .

[13]  J. Miletich,et al.  Inherited predisposition to thrombosis , 1993, Cell.

[14]  R. Woychik,et al.  Molecular characterization of the mouse agouti locus , 1992, Cell.

[15]  J. Seidman,et al.  Production of homozygous mutant ES cells with a single targeting construct , 1992, Molecular and cellular biology.

[16]  S. Kennel,et al.  Thrombomodulin is preferentially expressed in Balb/c lung microvessels. , 1992, The Journal of biological chemistry.

[17]  V L Bautch,et al.  Embryonic stem cell-derived cystic embryoid bodies form vascular channels: an in vitro model of blood vessel development. , 1992, Development.

[18]  R. Jackman,et al.  Expression of thrombomodulin by smooth muscle cells in culture: different effects of tumor necrosis factor and cyclic adenosine monophosphate on thrombomodulin expression by endothelial cells and smooth muscle cells in culture. , 1991, Blood.

[19]  H. Yamaguchi,et al.  Differential expression of fetomodulin and tissue plasminogen activator to characterize parietal endoderm differentiation of F9 embryonal carcinoma cells. , 1990, Developmental biology.

[20]  P. Majerus,et al.  A role for thrombomodulin in the pathogenesis of thrombin-induced thromboembolism in mice , 1988 .

[21]  T. Doetschman,et al.  Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies. , 1988, Development.

[22]  J. Griffin,et al.  The Frequency of Type I Heterozygous Protein S and Protein C Deficiency in 141 Unrelated Young Patients with Venous Thrombosis , 1988, Thrombosis and Haemostasis.

[23]  P. Vokonas,et al.  Aging-associated changes in indices of thrombin generation and protein C activation in humans. Normative Aging Study. , 1987, The Journal of clinical investigation.

[24]  M. Capecchi,et al.  Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells , 1987, Cell.

[25]  C. Haudenschild,et al.  Preservation of thrombomodulin antigen on vascular and extravascular surfaces. , 1987, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[26]  D. Beeler,et al.  Human thrombomodulin gene is intron depleted: nucleic acid sequences of the cDNA and gene predict protein structure and suggest sites of regulatory control. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[27]  H. Yamaguchi,et al.  Fetomodulin: marker surface protein of fetal development which is modulatable by cyclic AMP. , 1987, Developmental biology.

[28]  H. Dvorak,et al.  125I-fibrin deposition in contact sensitivity reactions in the mouse. Sensitivity of the assay for quantitating reactions after active or passive sensitization. , 1986, Journal of immunology.

[29]  C. Bell,et al.  Thrombomodulin is found on endothelium of arteries, veins, capillaries, and lymphatics, and on syncytiotrophoblast of human placenta , 1985, The Journal of cell biology.

[30]  R Kemler,et al.  The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. , 1985, Journal of embryology and experimental morphology.

[31]  E. Manseau,et al.  Pathogenesis of desmoplasia. I. Immunofluorescence identification and localization of some structural proteins of line 1 and line 10 guinea pig tumors and of healing wounds. , 1984, Journal of the National Cancer Institute.

[32]  J. Mcdonagh,et al.  High Resolution Electrophoretic Analysis of Human Fibrinogen and Its Crosslinked Intermediates , 1983, Thrombosis and Haemostasis.

[33]  C. Esmon,et al.  Activation of protein C in vivo. , 1982, The Journal of clinical investigation.

[34]  C. Esmon,et al.  Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. , 1981, Proceedings of the National Academy of Sciences of the United States of America.