Cellular movement is an essential component of normal developmental and reparative processes including embryogenesis, wound healing, tissue remodelling, and angiogenesis. Recent studies have identified a small group of proteins which appear to be specific regulators of cell mobility. One example is scatter factor (SF), a soluble fibroblast- derived protein activity which causes sheets of normal epithelial cells to scatter (spread and separate into single cells). We showed that bovine and human arterial smooth muscle cells release a factor similar to the human embryonic fibroblast-derived SF. We the utilized two quantitative assays of cell migration (a microcarrier bead assay developed in our laboratory and a modified Boyden chamber assay) to demonstrate that SF stimulates migration of arterial endothelium and epithelium, but not 3T3 or smooth muscle. While SF's in vivo functions are not yet known, these findings suggest that it might be involved in wound healing and vascular repair. Scattering and migration-stimulating activities were both heat sensitive, trypsin labile, and non-dialyzable (14 kd cutoff), and activities co-purified on standard and FPLC columns. Dose-response curves for stimulation of endothelial or epithelial migration were similar whether crude, partially purified or highly purified SF preparations were utilized. We purified the factor 500-1000 fold from serum-free conditioned medium from a high producer line of ras-transformed mouse 3T3 cells using cation exchange chromatography. FPLC purified fractions showed one major band at 62 kd on analysis by SDS-PAGE with silver- staining. We estimate that SF maximally stimulates migration of target cells at concentrations of about 0.5 nM. In this grant, we proposed to develop efficient methods for chromatographic purification of sufficient quantities of electrophoretically homogeneous SF to allow complete biochemical characterization of the protein, including N-terminal microsequencing. We will use the purified factor to generate monoclonals for development of immunoassays; in vivo detection, and further protein characterization. We will investigate SF's biologic activity on microvascular endothelium and its effects on the cytoskeletal organization of endothelial and epithelial cells. These studies will contribute to basic knowledge of cell movement and of blood vessel biology. They may also yield insights which will ultimately be useful in diagnosis and treatment of vascular disorders.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA050516-01
Application #
3195037
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1989-09-01
Project End
1992-08-31
Budget Start
1989-09-01
Budget End
1990-08-31
Support Year
1
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Rosen, E M; Goldberg, I D (1997) Regulation of angiogenesis by scatter factor. EXS 79:193-208
Joseph, A; Weiss, G H; Jin, L et al. (1995) Expression of scatter factor in human bladder carcinoma. J Natl Cancer Inst 87:372-7
Rosen, E M; Goldberg, I D (1995) Regulation of scatter factor (hepatocyte growth factor) production by tumor-stroma interaction. EXS 74:17-31
Rosen, E M; Goldberg, I D (1995) Scatter factor and angiogenesis. Adv Cancer Res 67:257-79
Rosen, E M; Knesel, J; Goldberg, I D et al. (1994) Scatter factor modulates the metastatic phenotype of the EMT6 mouse mammary tumor. Int J Cancer 57:706-14
Li, Y; Bhargava, M M; Joseph, A et al. (1994) Effect of hepatocyte growth factor/scatter factor and other growth factors on motility and morphology of non-tumorigenic and tumor cells. In Vitro Cell Dev Biol Anim 30A:105-10
Rosen, E M; Joseph, A; Jin, L et al. (1994) Regulation of scatter factor production via a soluble inducing factor. J Cell Biol 127:225-34
Rosen, E M; Zitnik, R J; Elias, J A et al. (1993) The interaction of HGF-SF with other cytokines in tumor invasion and angiogenesis. EXS 65:301-10
Jin, L; Pang, Y Y; Joseph, A et al. (1993) Rat placental hepatocyte growth factor/scatter factor: purification, characterization, and developmental regulation. Proc Soc Exp Biol Med 204:75-80
Grant, D S; Kleinman, H K; Goldberg, I D et al. (1993) Scatter factor induces blood vessel formation in vivo. Proc Natl Acad Sci U S A 90:1937-41

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