The specific objective of the proposed work is to develop, validate and apply methods for the determination of excreted alkylated purines as markers of DNA alkylation in vivo in humans. The following approaches will be used: i) novel methods will be used for the preparation of antibodies to low molecular weight alkyl purines. The antibodies will be used to develop quantitative immunoassay methods for the determination of urinary alkyl purines (3-methyl-, 3-ethyl- and 3-carboxy- methyladenine, and 7-ethyl- and 7-carboxymethylguanine). ii) Methods for the determination of urinary alkylated purines will be validated in animal models. The level and pattern of urinary excretion of 3-methyladenine will be correlated with methylation in both lymphocyte DNA and target tissue DNA following administration of dimethylnitrosamine. Similar experiments will be carried out for the urinary 3-ethyladenine and and 7-ethylguanine following administration f diethylnitrosamine, and 3-carboxymethyladenine and 7-carboxymethylguanine following N-nitrosoglycocholic acid administration. iii) Urinary alkylpurine determination will be incorporated into epidemiological studies where either cancer incidence in defined populations or individual cases are the endpoints. Methods will be developed for the identification and determination of cyclophosphamide DNA adducts in blood and urine samples of cancer patients receiving chemotherapy. Immunoassay and mass spectrometric methods will be developed for case-control studies in which characteristic DNA adducts will be evaluated as markers of risk for second cancer. The overall goal of this project is to develop validated, practical methods for monitoring human exposure to alkylating carcinogens based on the determination of urinary alkyl purines. In the case of iatrogenic second cancers following cyclophosphamide therapy the methods have potential for identifying DNA adducts indicative of the risk of developing second cancer. This would enable key ideas in molecular epidemiology to be validated and also offer the specific benefit of identifying modifications in therapy required to reduce the risk of second cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project--Cooperative Agreements (U01)
Project #
1U01CA048473-01
Application #
3549213
Study Section
(SRC)
Project Start
1988-07-01
Project End
1991-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
1
Fiscal Year
1988
Total Cost
Indirect Cost
Name
International Agency for Research on Cancer
Department
Type
DUNS #
279551881
City
Lyon
State
Country
France
Zip Code
69372
Prevost, V; Shuker, D E (1996) Cigarette smoking and urinary 3-alkyladenine excretion in man. Chem Res Toxicol 9:439-44
Shuker, D E; Bartsch, H (1994) Detection of human exposure to carcinogens by measurement of alkyl-DNA adducts using immunoaffinity clean-up in combination with gas chromatography-mass spectrometry and other methods of quantitation. Mutat Res 313:263-8
Shuker, D E; Prevost, V; Friesen, M D et al. (1993) Urinary markers for measuring exposure to endogenous and exogenous alkylating agents and precursors. Environ Health Perspect 99:33-7
Shuker, D E; Farmer, P B (1992) Relevance of urinary DNA adducts as markers of carcinogen exposure. Chem Res Toxicol 5:450-60
Shuker, D E; Friesen, M D; Garren, L et al. (1991) A rapid gas chromatography-mass spectrometry method for the determination of urinary 3-methyladenine: application in human subjects. IARC Sci Publ :102-6
Friesen, M D; Garren, L; Prevost, V et al. (1991) Isolation of urinary 3-methyladenine using immunoaffinity columns prior to determination by low-resolution gas chromatography-mass spectrometry. Chem Res Toxicol 4:102-6
Bartsch, H; Shuker, D E; Ohshima, H (1991) Human nitrosamine exposure: recent dosimetry methods and applications. Prog Clin Biol Res 372:197-204
Bartsch, H; Ohshima, H; Shuker, D E et al. (1990) Exposure of humans to endogenous N-nitroso compounds: implications in cancer etiology. Mutat Res 238:255-67
Prevost, V; Shuker, D E; Bartsch, H et al. (1990) The determination of urinary 3-methyladenine by immunoaffinity chromatography-monoclonal antibody-based ELISA: use in human biomonitoring studies. Carcinogenesis 11:1747-51
Shuker, D E (1989) Detection of adducts arising from human exposure to N-nitroso compounds. Cancer Surv 8:475-87

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