Endothelium has a central role in the regulation of vascular permeability and the coagulation mechanism. In the prethrombotic state associated with certain malignancies, the coagulation mechanism and vascular permeability appear to be altered. This proposal examines the effects of an apparently novel polypeptide, isolated from murine meth A tumor cells, on barrier function and cell surface coagulant properties of the endothelial cell. Preliminary studies, which resulted in the identification of an isolation procedure for this 24,000 dalton protein from tumor-conditioned medium (meth A factor), will be optimized and scaled-up to produce sufficient material for further biochemical characterization, production of antisera and physiology studies in vitro and in vivo. The human counterpart of the murine factor, which has been partially purified from a melanoma cell line, will be completed. Using purified meth A factor and cultured endothelium (micro- and macrovascular), pilot studies demonstrating modulation of endothelial cell coagulant and barrier function by this mediator will be extended, and the underlying mechanisms identified: partially purified or purified meth A factor suppresses two anticoagulant mechanisms of endothelium (the protein C/protein S and fibrinolytic pathways), enhances induction of the procoagulant cofactor tissue factor in response to TNF, and increases monolayer permeability. In vivo studies have shown a selective procoagulant response in the tumor vasculature of meth A sarcomas following the administration of low concentrations of TNF, which we hypothesize is related to the meth A factor. Furthermore, partially purified meth A factor augments the increase in vascular permeability observed in the presence of TNF in vivo. These observations will be followed up with systematic studies in rats to define the effects of meth A factor on coagulant function and vascular blood flow/permeability. Activation of coagulation will be monitored by assays for fibrinopeptide A and fragment 1+2, and direct examination for the presence of intravascular fibrin. Measurements made by intravital microscopy will define blood flow and transvascular exchange of macromolecules. The presence of tumor factor in malignant tissues will be assessed using our immunological reagents. These specific plans are directed toward our long-term objective: understanding the role of endothelium in the pathogenesis of the prethrombotic state.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL042833-02
Application #
3361146
Study Section
Special Emphasis Panel (SRC (PM))
Project Start
1989-03-01
Project End
1993-12-31
Budget Start
1990-01-01
Budget End
1990-12-31
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Schwarz, M A; Kandel, J; Brett, J et al. (1999) Endothelial-monocyte activating polypeptide II, a novel antitumor cytokine that suppresses primary and metastatic tumor growth and induces apoptosis in growing endothelial cells. J Exp Med 190:341-54
Niitsu, Y; Hori, O; Yamaguchi, A et al. (1999) Exposure of cultured primary rat astrocytes to hypoxia results in intracellular glucose depletion and induction of glycolytic enzymes. Brain Res Mol Brain Res 74:26-34
Bohrer, H; Qiu, F; Zimmermann, T et al. (1997) Role of NFkappaB in the mortality of sepsis. J Clin Invest 100:972-85
Zhang, Y; Deng, Y; Wendt, T et al. (1996) 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. J Clin Invest 97:2213-24
Hori, O; Matsumoto, M; Kuwabara, K et al. (1996) Exposure of astrocytes to hypoxia/reoxygenation enhances expression of glucose-regulated protein 78 facilitating astrocyte release of the neuroprotective cytokine interleukin 6. J Neurochem 66:973-9
Hori, O; Yan, S D; Ogawa, S et al. (1996) The receptor for advanced glycation end-products has a central role in mediating the effects of advanced glycation end-products on the development of vascular disease in diabetes mellitus. Nephrol Dial Transplant 11 Suppl 5:13-6
Marvin, M R; Libutti, S K; Kayton, M et al. (1996) A novel tumor-derived mediator that sensitizes cytokine-resistant tumors to tumor necrosis factor. J Surg Res 63:248-55
Lawson, C A; Smerling, A J; Naka, Y et al. (1995) Selective reduction of PVR by inhalation of a cGMP analogue in a porcine model of pulmonary hypertension. Am J Physiol 268:H2056-62
Yellin, M J; Brett, J; Baum, D et al. (1995) Functional interactions of T cells with endothelial cells: the role of CD40L-CD40-mediated signals. J Exp Med 182:1857-64
Yuzawa, Y; Brett, J; Fukatsu, A et al. (1995) Interaction of antibody with Forssman antigen in guinea pigs. A mechanism of adaptation to antibody- and complement-mediated injury. Am J Pathol 146:1260-72

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