My long-term goal is to identify the genes and gen products responsible for the proliferation and malignant transformation of human melanocytes. My approach is based on knowledge concerning the growth factor requirements of normal and malignant human melanocytes in culture. Normal human melanocytes, unlike pigment cells from metastatic melanomas, do not survive in culture, neither in serum-supplemented nor in defined medium. Until recently, normal human melanocytes could proliferate well in culture only in the presence of 12-0-tetradecanoyl-phorbol-13-acetate (TPA) in synergy with substances that increase intracellular levels of cyclic-adenosine-monophosphate (cAMP). We demonstrated that mitogenic activity in normal tissues and melanoma cell extracts was immunologically related to basic fibroblast growth factor (bFGF) and that bFGF was the only mitogen for melanocytes that could substitute for TPA. It is thus possible that abnormal production of bFGF has a role in the uncontrolled growth of transformed melanocytes. Other gene products might be those conferring independence from cAMP stimulators.
My specific aims are therefore: I. To determine at which stage of malignant transformation melanomas becomes independent of and/or produce bFGF, and what is the mechanism of bFGF induction. II. To find out whether the genes that confer independence form TOA, bFGF, and stimulators of cAMP to melanomas are the genes for bFGF and cAMP-dependent protein kinases. III. To identify the receptor for bFGF in melanocytes and to compare its expression in normal and malignant melanocytes. IV. To compare the expression of cAMP-dependent protein kinases in normal and malignant melanocytes. The techniques to be employed are cultured normal and malignant melanocytes, antibodies to bFGF and subunits of protein kinase A, Northern blot analysis with cDNA probes to mRNAs of bFGF and subunits of protein kinase A, cellular DNA transfection, and transfection with specific constructs encoding bFGF and the catalytic subunit of protein kinase A.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA044542-04
Application #
3458011
Study Section
Pathology B Study Section (PTHB)
Project Start
1988-03-01
Project End
1993-02-28
Budget Start
1991-03-01
Budget End
1992-02-29
Support Year
4
Fiscal Year
1991
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
Halaban, R; Moellmann, G (1993) White mutants in mice shedding light on humans. J Invest Dermatol 100:176S-185S
Halaban, R; Tyrrell, L; Longley, J et al. (1993) Pigmentation and proliferation of human melanocytes and the effects of melanocyte-stimulating hormone and ultraviolet B light. Ann N Y Acad Sci 680:290-301
Zakut, R; Perlis, R; Eliyahu, S et al. (1993) KIT ligand (mast cell growth factor) inhibits the growth of KIT-expressing melanoma cells. Oncogene 8:2221-9
Halaban, R; Rubin, J S; White, W (1993) met and HGF-SF in normal melanocytes and melanoma cells. EXS 65:329-39
Gitay-Goren, H; Halaban, R; Neufeld, G (1993) Human melanoma cells but not normal melanocytes express vascular endothelial growth factor receptors. Biochem Biophys Res Commun 190:702-8
Halaban, R (1993) Molecular correlates in the progression of normal melanocytes to melanomas. Semin Cancer Biol 4:171-81
Longley Jr, B J; Morganroth, G S; Tyrrell, L et al. (1993) Altered metabolism of mast-cell growth factor (c-kit ligand) in cutaneous mastocytosis. N Engl J Med 328:1302-7
Funasaka, Y; Boulton, T; Cobb, M et al. (1992) c-Kit-kinase induces a cascade of protein tyrosine phosphorylation in normal human melanocytes in response to mast cell growth factor and stimulates mitogen-activated protein kinase but is down-regulated in melanomas. Mol Biol Cell 3:197-209
Halaban, R; Rubin, J S; Funasaka, Y et al. (1992) Met and hepatocyte growth factor/scatter factor signal transduction in normal melanocytes and melanoma cells. Oncogene 7:2195-206
Halaban, R; Fan, B; Ahn, J et al. (1992) Growth factors, receptor kinases, and protein tyrosine phosphatases in normal and malignant melanocytes. J Immunother (1991) 12:154-61

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