The objective of this application is to further characterize the molecular and biological events that occur following genotoxin exposure of cells and tissues derived from human fetal brain (glial and neuronal cells), kidney (proximal tubular and glomerular epithelial cells), and skin (fibroblastic and epithelial cells). DNA damage and repair will be characterized and quantitated as a function of DNA modification, organ and cell type as related to individual, organ and cell sensitivity. UV radiation and nitrosoureas will be used to represent direct acting genotoxins which induce well characterized bulky (pyrimidine dimers) and non-bulky (N7 and 06-alkylguaine, 04 and 02-alkylthymine, N3-alkyladenine, phhosphotriester, and apurinic sites) lesions in the DNA. Studies will be done, when possible, simultaneously with both fresh human fetal tissue and cultured cells derived from the same specimen. The inhibition, saturability and inducibility of the repair enzymes with related and unrelated agents either separately or in combination with different agents (i.e., UV, alkylator, AAAF) will be investigated in cell culture as a function of dose and time of exposure to genotoxin and in specific stages of the cell cycle, e.g. G1 vs S phase. Indiviudal cell and organ sensitivity will be measured by the dose and time dependent changes in cell morphology and growth rate, cytotoxicity, anchorage independent growth and nucleic acid and protein synthesis. The studies of this renewal research grant are designed to provide new information and insights into the mechanisms of genotoxicity on cellular function in relation to human organ and cell type. The long term goal of this research program is to interrelate the molecular events of genotoxic DNA damage and DNA repair of specific DNA modifications to the biological parameters in humans in vivo and in vitro.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
2R01ES003101-04
Application #
3250237
Study Section
Toxicology Study Section (TOX)
Project Start
1983-06-20
Project End
1991-05-31
Budget Start
1986-06-20
Budget End
1987-05-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Ohio State University
Department
Type
Schools of Medicine
DUNS #
098987217
City
Columbus
State
OH
Country
United States
Zip Code
43210
Tong, H H; Park, J H; Brady, T et al. (1997) Molecular characterization of mutations in the hprt gene of normal human skin keratinocytes treated with N-ethyl-N-nitrosourea: influence of O6-alkylguanine alkyltransferase. Environ Mol Mutagen 29:168-79
Wani, G; Wani, A A; D'Ambrosio, S M (1992) In situ hybridization of human kidney tissue reveals cell-type-specific expression of the O6-methylguanine-DNA methyltransferase gene. Carcinogenesis 13:463-8
Gibson-D'Ambrosio, R E; D'Ambrosio, S M (1992) Long-term culture of normal human kidney glomerular cells. In Vitro Cell Dev Biol 28A:471-4
D'Ambrosio, S M; Wani, G; Samuel, M et al. (1990) Repair of O6-methylguanine damage in normal human tissues. Basic Life Sci 53:397-416
Wani, A A; Wani, G; D'Ambrosio, S M (1990) A human DNA repair activity specific for O4-ethylthymine: identification and partial characterization. Carcinogenesis 11:1419-24
Wani, A A; Wani, G; D'Ambrosio, S M (1990) Repair of O4-alkylthymine damage in human cells. Basic Life Sci 53:417-35
Wani, G; Wani, A A; Gibson-D'Ambrosio, R et al. (1989) Absence of DNA damage-mediated induction of human methyltransferase specific for precarcinogenic O6-methylguanine. Teratog Carcinog Mutagen 9:259-72
Milo, G E; D'Ambrosio, S; Kun, E (1989) Benzamide prevention of ultraviolet radiation-induced transformation as measured by anchorage-independent growth and the absence of correlation with thymidine dimer formation and DNA repair. Teratog Carcinog Mutagen 9:167-76
Wani, A A; D'Ambrosio, S M; Alvi, N K (1987) Quantitation of pyrimidine dimers by immunoslot blot following sublethal UV-irradiation of human cells. Photochem Photobiol 46:477-82
Chang, C C; Trosko, J E; el-Fouly, M H et al. (1987) Contact insensitivity of a subpopulation of normal human fetal kidney epithelial cells and of human carcinoma cell lines. Cancer Res 47:1634-45

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