Our specific aims are to analyze whether transfection of vascular endothelial cells with bFGF cDNA expression vector could lead to autonomous proliferation of vascular endothelial cells and their transformation into neoplastic cells. We will first analyze whether bFGF would act intracellularly (as most oncogene products do) or need to be released by the cells in order to interact with exposed cell surface receptors. We will also analyze its cellular localization, in order to determine whether it could ultimately become associated with the nuclear, cytosol, or plasma membrane fraction. Since cell transformation can be the direct result of oncogene expression, we will analyze in transfected vascular endothelial cells whether constitutive expression of c fos, c myc and c ras oncogenes can be observed. Whether the bFGF gene product could be released from the cells in association with ECM components such as heparan sulfate GAG, for which the mitogen should have a high affinity, will be assessed. The effect of TGFB and TNF on the growth of parental and transfected cells in soft agar, as well as PA expression, will be analyzed in order to determine whether they could have effects similar to those reported when cells are grown on solid substrates. Monoclonal antibodies against the FGF receptor will be obtained and used for the development of sensitive RIA for the FGF receptor, and studies of its expression. They will also be used to isolate FGF receptors by immunoaffinity chromatography. The FGF receptor will then be characterized structurally, and the information gained will be used to develop sepecific oligonucleotide probes, which will be used for the cloning of 2 FGF receptor cDNA.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL020197-14
Application #
3336078
Study Section
Pathology B Study Section (PTHB)
Project Start
1976-12-01
Project End
1990-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
14
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Gospodarowicz, D; Abraham, J A; Schilling, J (1989) Isolation and characterization of a vascular endothelial cell mitogen produced by pituitary-derived folliculo stellate cells. Proc Natl Acad Sci U S A 86:7311-5
Schweigerer, L; Ferrara, N; Haaparanta, T et al. (1988) Basic fibroblast growth factor: expression in cultured cells derived from corneal endothelium and lens epithelium. Exp Eye Res 46:71-80
Gospodarowicz, D; Ferrara, N; Haaparanta, T et al. (1988) Basic fibroblast growth factor: expression in cultured bovine vascular smooth muscle cells. Eur J Cell Biol 46:144-51
Neufeld, G; Gospodarowicz, D (1988) Identification of the fibroblast growth factor receptor in human vascular endothelial cells. J Cell Physiol 136:537-42
Adashi, E Y; Resnick, C E; Croft, C S et al. (1988) Basic fibroblast growth factor as a regulator of ovarian granulosa cell differentiation: a novel non-mitogenic role. Mol Cell Endocrinol 55:7-14
Neufeld, G; Gospodarowicz, D (1987) Protamine sulfate inhibits mitogenic activities of the extracellular matrix and fibroblast growth factor, but potentiates that of epidermal growth factor. J Cell Physiol 132:287-94
Schweigerer, L; Malerstein, B; Neufeld, G et al. (1987) Basic fibroblast growth factor is synthesized in cultured retinal pigment epithelial cells. Biochem Biophys Res Commun 143:934-40
Schweigerer, L; Neufeld, G; Gospodarowicz, D (1987) Basic fibroblast growth factor as a growth inhibitor for cultured human tumor cells. J Clin Invest 80:1516-20
Schweigerer, L; Neufeld, G; Gospodarowicz, D (1987) Basic fibroblast growth factor is present in cultured human retinoblastoma cells. Invest Ophthalmol Vis Sci 28:1838-43
Massoglia, S L; Kenney, J S; Gospodarowicz, D J (1987) Characterization of murine monoclonal antibodies directed against basic fibroblast growth factor. J Cell Physiol 132:531-7

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