We propose to develop monoclonal antibody-based procedures for molecular dosimetry of depurinating DNA adducts of benzo[alpha]pyrene (BP) and dibenzo[alpha,l]pyrene (DB[alpha,l]P), carcinogens viewed as indicators of exposure to polycyclic aromatic hydrocarbons (PAH) and risk of cancer, and catechol estrogens (CE), putative endogenous procarcinogens. Recent data support an essential contribution of depurinating adducts to cancer initiation by both PAH and CE. Since these adducts are frequently the predominant ones formed, are rapidly lost from the DNA and are excreted in urine, they are potentially useful biomarkers of chemical-specific DNA damage and risk for cancer. This project encompasses the production of monoclonal antibodies (MABs) specific for BP=6=C8Gua(a major depurinating adduct of BP;
Aim #1), syn-DB[alpha,l]PDE-14-N7Ade and DB[alpha,l]P-10- N3Ade (major depurinating adducts of DB[alpha,l]P;
Aim #2), and 4-OHE1- (1&2)}-N7Gua and 4-OHE2-(1&2)-N7Gua (predominant depurinating adducts of CE;
Aim #3). The specific affinities of these MAbs will be determined by competitive enzyme-linked immunosorbent assay (ELISA). In the early stages of development of the proposed molecular dosimeters, MAbs of high specific affinity will be incorporated into competitive ELISAs and immunoaffinity HPLC (1A/HPLC) for quantitation of each adduct added to urine collected from normal rats and humans. Dose-response studies will compare urinary adducts, measured by competitive ELISA or IA/HPLC, with doses of PAH or CE given to rats. Transitional epidemiological studies will apply the MAb- based dosimeters to quantitation of BP adducts in the urine of smokers and nonsmokers, and DB[alpha,l]P adducts in the urine of individuals who are exposed to byproducts of smoky coal combustion and are at high risk for lung cancer. These studies will provide molecular dosimetric procedures needed for prospective epidemiological studies and determination of individual risk for both PAH- and CE-induced cancers.

National Institute of Health (NIH)
National Cancer Institute (NCI)
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University of Nebraska Medical Center
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Cavalieri, Ercole; Saeed, Muhammad; Zahid, Muhammad et al. (2012) Mechanism of DNA depurination by carcinogens in relation to cancer initiation. IUBMB Life 64:169-79
Zahid, Muhammad; Saeed, Muhammad; Rogan, Eleanor G et al. (2010) Benzene and dopamine catechol quinones could initiate cancer or neurogenic disease. Free Radic Biol Med 48:318-24
Saeed, Muhammad; Rogan, Eleanor; Cavalieri, Ercole (2009) Mechanism of metabolic activation and DNA adduct formation by the human carcinogen diethylstilbestrol: the defining link to natural estrogens. Int J Cancer 124:1276-84
Saeed, Muhammad; Higginbotham, Sheila; Gaikwad, Nilesh et al. (2009) Depurinating naphthalene-DNA adducts in mouse skin related to cancer initiation. Free Radic Biol Med 47:1075-81
Lu, Fang; Zahid, Muhammad; Wang, Cheng et al. (2008) Resveratrol prevents estrogen-DNA adduct formation and neoplastic transformation in MCF-10F cells. Cancer Prev Res (Phila Pa) 1:135-45
Gaikwad, Nilesh W; Yang, Li; Muti, Paola et al. (2008) The molecular etiology of breast cancer: evidence from biomarkers of risk. Int J Cancer 122:1949-57
Chakravarti, Dhrubajyoti; Venugopal, Divya; Mailander, Paula C et al. (2008) The role of polycyclic aromatic hydrocarbon-DNA adducts in inducing mutations in mouse skin. Mutat Res 649:161-78
Zahid, Muhammad; Saeed, Muhammad; Lu, Fang et al. (2007) Inhibition of catechol-O-methyltransferase increases estrogen-DNA adduct formation. Free Radic Biol Med 43:1534-40
Gaikwad, Nilesh W; Rogan, Eleanor G; Cavalieri, Ercole L (2007) Evidence from ESI-MS for NQO1-catalyzed reduction of estrogen ortho-quinones. Free Radic Biol Med 43:1289-98
Zahid, Muhammad; Gaikwad, Nilesh W; Rogan, Eleanor G et al. (2007) Inhibition of depurinating estrogen-DNA adduct formation by natural compounds. Chem Res Toxicol 20:1947-53

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