Oral Cell DNA Adducts in Smokers

Using high resolution mass spectrometry, quantify known DNA adducts in oral mucosa cells of
100 smokers from each ethnic group - Native Hawaiians, Whites, and Japanese Americans. DNA
adducts of tobacco-specific compounds, formaldehyde, and acrolein will be quantified.

2. Analyze the urine of 100 smokers and 100 non-smokers from each of these groups for
mercapturic acids of acrolein and crotonaldehyde, the F2-isoprostane 8-iso-PGF-2α, a
biomarker of oxidative damage, and the prostaglandin E2 metabolite PGEM, a biomarker of
inflammation, as well as total nicotine equivalents and total NNAL (smokers only). These data
will provide critical information relevant to the high risk of Native Hawaiians for lung
cancer, and in relationship to the DNA adduct measurements of Specific Aim 1.

3. Use newly developed high resolution mass spectrometric, high throughput DNA adductomic
approach to screen for known and unknown DNA adducts to identify a comprehensive DNA
modification signature derived from cigarette smoking.

1. Using a targeted DNA adductomic method, screen for multiple DNA modifications
simultaneously. The adducts analyzed in Specific Aim 1 will be added to a list including
endogenous, 1,3-butadiene-derived (in collaboration with Project 3), aldehyde-derived
and nitrosamine or alkylating agent-derived DNA adducts. In parallel, apply DNA
adductomic methods in an untargeted mode to investigate the presence of previously
unknown DNA adducts. The unknown adducts detected by the untargeted analysis will be
included in the list of targeted DNA adducts to ultimately obtain a sensitive method to
assess overall DNA damage resulting from cigarette smoking.

2. The list of DNA adducts created in Specific Aim 3a will be applied to investigate
differences in the DNA damage signature in selected samples analyzed in Specific Aim 1.

Inclusion Criteria:

- currently smoke at least 5 cigarettes per day, self-identified Japanese Americans,
European Americans, or Native Hawaiians.

Exclusion Criteria:

- no current use of other nicotine containing products, no acute or uncontrolled medical
or psychiatric conditions, no greater than 14 alcoholic drinks per week, not pregnant,
not breastfeeding, not currently taking any medications that affect relevant metabolic
enzymes, no active infection.