Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 May 15;97(10):2213–2224. doi: 10.1172/JCI118662

Intravenous somatic gene transfer with antisense tissue factor restores blood flow by reducing tumor necrosis factor-induced tissue factor expression and fibrin deposition in mouse meth-A sarcoma.

Y Zhang 1, Y Deng 1, T Wendt 1, B Liliensiek 1, A Bierhaus 1, J Greten 1, W He 1, B Chen 1, V Hach-Wunderle 1, R Waldherr 1, R Ziegler 1, D Männel 1, D M Stern 1, P P Nawroth 1
PMCID: PMC507300  PMID: 8636400

Abstract

Fibrin is deposited on the endothelial cell surface in the vasculature of murine methylcholanthrene A-induced sarcomas after injection of tumor necrosis factor (TNF). Capillary endothelial cells of the tumor vascular bed become positive for tissue factor after TNF injection, based on immunocytochemistry and in situ hybridization. Intravascular clot formation was not dependent on tissue factor derived from tumor cells, since in vessels of tumors not expressing tissue factor, TNF also induced fibrin/fibrinogen deposition. However, the time course of fibrin/fibrinogen deposition after TNF differed in tumors expressing no, little, or greater amounts of tissue factor. Fibrin/fibrinogen deposition was more rapid in tumors in which the neoplastic cells expressed tissue factor than in tumors not expressing tissue factor. In the tumors not expressing tissue factor, activation of coagulation was dependent on TNF-induced synthesis of tissue factor by host cells, i.e., endothelium or monocytes/macrophages. Intravenous somatic gene transfer with tissue factor cDNA in the antisense orientation (but not sense or vector alone) reduced intravascular fibrin/fibrinogen deposition and restored blood flow to the tumor, showing that de novo tissue factor expression is central in TNF-induced activation of the coagulation mechanism.

Full Text

The Full Text of this article is available as a PDF (644.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alam J., Cook J. L. Reporter genes: application to the study of mammalian gene transcription. Anal Biochem. 1990 Aug 1;188(2):245–254. doi: 10.1016/0003-2697(90)90601-5. [DOI] [PubMed] [Google Scholar]
  2. Andree H. A., Nemerson Y. Tissue factor: regulation of activity by flow and phospholipid surfaces. Blood Coagul Fibrinolysis. 1995 May;6(3):189–197. [PubMed] [Google Scholar]
  3. Bach R. R. Initiation of coagulation by tissue factor. CRC Crit Rev Biochem. 1988;23(4):339–368. doi: 10.3109/10409238809082548. [DOI] [PubMed] [Google Scholar]
  4. Beutler B., Milsark I. W., Cerami A. C. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985 Aug 30;229(4716):869–871. doi: 10.1126/science.3895437. [DOI] [PubMed] [Google Scholar]
  5. Bevilacqua M. P., Pober J. S., Majeau G. R., Fiers W., Cotran R. S., Gimbrone M. A., Jr Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the actions of interleukin 1. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4533–4537. doi: 10.1073/pnas.83.12.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bierhaus A., Zhang Y., Deng Y., Mackman N., Quehenberger P., Haase M., Luther T., Müller M., Böhrer H., Greten J. Mechanism of the tumor necrosis factor alpha-mediated induction of endothelial tissue factor. J Biol Chem. 1995 Nov 3;270(44):26419–26432. doi: 10.1074/jbc.270.44.26419. [DOI] [PubMed] [Google Scholar]
  7. Boulter C. A., Wagner E. F. A universal retroviral vector for efficient constitutive expression of exogenous genes. Nucleic Acids Res. 1987 Sep 11;15(17):7194–7194. doi: 10.1093/nar/15.17.7194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brett J., Gerlach H., Nawroth P., Steinberg S., Godman G., Stern D. Tumor necrosis factor/cachectin increases permeability of endothelial cell monolayers by a mechanism involving regulatory G proteins. J Exp Med. 1989 Jun 1;169(6):1977–1991. doi: 10.1084/jem.169.6.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Camussi G., Turello E., Bussolino F., Baglioni C. Tumor necrosis factor alters cytoskeletal organization and barrier function of endothelial cells. Int Arch Allergy Appl Immunol. 1991;96(1):84–91. doi: 10.1159/000235539. [DOI] [PubMed] [Google Scholar]
  10. Clark M. A., Chen M. J., Crooke S. T., Bomalaski J. S. Tumour necrosis factor (cachectin) induces phospholipase A2 activity and synthesis of a phospholipase A2-activating protein in endothelial cells. Biochem J. 1988 Feb 15;250(1):125–132. doi: 10.1042/bj2500125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clauss M., Gerlach M., Gerlach H., Brett J., Wang F., Familletti P. C., Pan Y. C., Olander J. V., Connolly D. T., Stern D. Vascular permeability factor: a tumor-derived polypeptide that induces endothelial cell and monocyte procoagulant activity, and promotes monocyte migration. J Exp Med. 1990 Dec 1;172(6):1535–1545. doi: 10.1084/jem.172.6.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Clauss M., Murray J. C., Vianna M., de Waal R., Thurston G., Nawroth P., Gerlach H., Bach R., Familletti P. C., Stern D. A polypeptide factor produced by fibrosarcoma cells that induces endothelial tissue factor and enhances the procoagulant response to tumor necrosis factor/cachectin. J Biol Chem. 1990 Apr 25;265(12):7078–7083. [PubMed] [Google Scholar]
  13. Conway E. M., Bach R., Rosenberg R. D., Konigsberg W. H. Tumor necrosis factor enhances expression of tissue factor mRNA in endothelial cells. Thromb Res. 1989 Feb 1;53(3):231–241. doi: 10.1016/0049-3848(89)90098-4. [DOI] [PubMed] [Google Scholar]
  14. Drake T. A., Cheng J., Chang A., Taylor F. B., Jr Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol. 1993 May;142(5):1458–1470. [PMC free article] [PubMed] [Google Scholar]
  15. Drake T. A., Morrissey J. H., Edgington T. S. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol. 1989 May;134(5):1087–1097. [PMC free article] [PubMed] [Google Scholar]
  16. Echtenacher B., Falk W., Männel D. N., Krammer P. H. Requirement of endogenous tumor necrosis factor/cachectin for recovery from experimental peritonitis. J Immunol. 1990 Dec 1;145(11):3762–3766. [PubMed] [Google Scholar]
  17. Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  19. Hartzell S., Ryder K., Lanahan A., Lau L. F., Nathan D. A growth factor-responsive gene of murine BALB/c 3T3 cells encodes a protein homologous to human tissue factor. Mol Cell Biol. 1989 Jun;9(6):2567–2573. doi: 10.1128/mcb.9.6.2567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kao J., Ryan J., Brett G., Chen J., Shen H., Fan Y. G., Godman G., Familletti P. C., Wang F., Pan Y. C. Endothelial monocyte-activating polypeptide II. A novel tumor-derived polypeptide that activates host-response mechanisms. J Biol Chem. 1992 Oct 5;267(28):20239–20247. [PubMed] [Google Scholar]
  21. Kaplow I. S. Letter: Substitute for benzidine in myeloperoxidase stains. Am J Clin Pathol. 1975 Mar;63(3):451–451. doi: 10.1093/ajcp/63.3.451. [DOI] [PubMed] [Google Scholar]
  22. Keller G., Paige C., Gilboa E., Wagner E. F. Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature. 1985 Nov 14;318(6042):149–154. doi: 10.1038/318149a0. [DOI] [PubMed] [Google Scholar]
  23. Mackman N., Sawdey M. S., Keeton M. R., Loskutoff D. J. Murine tissue factor gene expression in vivo. Tissue and cell specificity and regulation by lipopolysaccharide. Am J Pathol. 1993 Jul;143(1):76–84. [PMC free article] [PubMed] [Google Scholar]
  24. Maynard J. R., Heckman C. A., Pitlick F. A., Nemerson Y. Association of tissue factor activity with the surface of cultured cells. J Clin Invest. 1975 Apr;55(4):814–824. doi: 10.1172/JCI107992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nawroth P. P., Bank I., Handley D., Cassimeris J., Chess L., Stern D. Tumor necrosis factor/cachectin interacts with endothelial cell receptors to induce release of interleukin 1. J Exp Med. 1986 Jun 1;163(6):1363–1375. doi: 10.1084/jem.163.6.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nawroth P. P., Stern D. M., Kisiel W., Bach R. Cellular requirements for tissue factor generation by bovine aortic endothelial cells in culture. Thromb Res. 1985 Dec 1;40(5):677–691. doi: 10.1016/0049-3848(85)90305-6. [DOI] [PubMed] [Google Scholar]
  27. Nawroth P. P., Stern D. M. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med. 1986 Mar 1;163(3):740–745. doi: 10.1084/jem.163.3.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nawroth P., Handley D., Matsueda G., De Waal R., Gerlach H., Blohm D., Stern D. Tumor necrosis factor/cachectin-induced intravascular fibrin formation in meth A fibrosarcomas. J Exp Med. 1988 Aug 1;168(2):637–647. doi: 10.1084/jem.168.2.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nilsson B. O., Larsson A. Intrasplenic immunization with minute amounts of antigen. Immunol Today. 1990 Jan;11(1):10–12. doi: 10.1016/0167-5699(90)90004-s. [DOI] [PubMed] [Google Scholar]
  30. Ow D. W., DE Wet J. R., Helinski D. R., Howell S. H., Wood K. V., Deluca M. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science. 1986 Nov 14;234(4778):856–859. doi: 10.1126/science.234.4778.856. [DOI] [PubMed] [Google Scholar]
  31. Ritthaler U., Deng Y., Zhang Y., Greten J., Abel M., Sido B., Allenberg J., Otto G., Roth H., Bierhaus A. Expression of receptors for advanced glycation end products in peripheral occlusive vascular disease. Am J Pathol. 1995 Mar;146(3):688–694. [PMC free article] [PubMed] [Google Scholar]
  32. Ryan J., Brett J., Tijburg P., Bach R. R., Kisiel W., Stern D. Tumor necrosis factor-induced endothelial tissue factor is associated with subendothelial matrix vesicles but is not expressed on the apical surface. Blood. 1992 Aug 15;80(4):966–974. [PubMed] [Google Scholar]
  33. Scharfmann R., Axelrod J. H., Verma I. M. Long-term in vivo expression of retrovirus-mediated gene transfer in mouse fibroblast implants. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4626–4630. doi: 10.1073/pnas.88.11.4626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Singer R. H., Ward D. C. Actin gene expression visualized in chicken muscle tissue culture by using in situ hybridization with a biotinated nucleotide analog. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7331–7335. doi: 10.1073/pnas.79.23.7331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Taylor F. B., Jr, Chang A., Ruf W., Morrissey J. H., Hinshaw L., Catlett R., Blick K., Edgington T. S. Lethal E. coli septic shock is prevented by blocking tissue factor with monoclonal antibody. Circ Shock. 1991 Mar;33(3):127–134. [PubMed] [Google Scholar]
  37. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss H. J., Lages B. Evidence for tissue factor-dependent activation of the classic extrinsic coagulation mechanism in blood obtained from bleeding time wounds. Blood. 1988 Mar;71(3):629–635. [PubMed] [Google Scholar]
  39. Wilcox J. N., Smith K. M., Schwartz S. M., Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2839–2843. doi: 10.1073/pnas.86.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Zhang Y. M., Bachmann S., Hemmer C., van Lunzen J., von Stemm A., Kern P., Dietrich M., Ziegler R., Waldherr R., Nawroth P. P. Vascular origin of Kaposi's sarcoma. Expression of leukocyte adhesion molecule-1, thrombomodulin, and tissue factor. Am J Pathol. 1994 Jan;144(1):51–59. [PMC free article] [PubMed] [Google Scholar]
  41. Zhang Y., Deng Y., Luther T., Müller M., Ziegler R., Waldherr R., Stern D. M., Nawroth P. P. Tissue factor controls the balance of angiogenic and antiangiogenic properties of tumor cells in mice. J Clin Invest. 1994 Sep;94(3):1320–1327. doi: 10.1172/JCI117451. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES