The broad, long-term objective of this application is to establish chemical markers of internal exposure and genetic damage through biomonitoring DNA adducts to assess the risk of carcinogens. Human risk assessment has been severely impeded by the paucity of analytical methods that unambiguously identify and quantitate DNA adducts in vivo, and by the inability to easily obtain relevant tissues to assess cancer development. We hypothesize that the recent advances in sensitivity of liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) instrumentation allow for the identification and quantification of DNA adducts in exfoliated buccal cells of individuals exposed to tobacco smoke and diet-related carcinogens. The successful use of buccal cells as a reliable tissue to measure DNA adducts requires optimization of the analytical MS methods and the establishment of standardized methods to isolate human buccal cell DNA. To address this hypothesis, the specific aims are: 1) To establish ultra-trace LC/ESI-MS methods to measure DNA adducts of suspected human carcinogens found in tobacco smoke that include: 4-methylnitrosamino-1-(3-pyridyl)-1-butanone, benzo(a)pyrene, 2-amino-a-carboline, 2-amino-3-methyl-a-carboline, 4-aminobiphenyl; and abundant heterocyclic aromatic amine (HAAs) carcinogens formed in grilled meats; 2) Cytobrush and mouthwash rinsing techniques will be compared to isolate buccal cells that maximize the recovery of human DNA and minimize contaminating bacterial DNA. The standardization of the buccal cell DNA isolation method will permit reliable assessment of genotoxin exposure and DNA damage; and 3) Recruit volunteers that include: smokers, non-smokers, and meat-eaters and probe for buccal cell DNA adducts by LC/ESI-MS. Adduct formation will be correlated to salivary nicotine levels for tobacco exposure and HAAs in the diet. The results of this research are expected to show that quantitative LC/ESI-MS analysis of buccal cell DNA adducts may be used to assess damage posed by genotoxins, and reduce the invasiveness of risk assessment procedures by validation of exfoliated cell specimens. The impact of this research on public health is the development of new analytical tools that may be used for biomonitoring and risk assessment, which examine exposure to genotoxicants, assess gene-environment interactions and determinants of cancer, or chemopreventive efficacy. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21ES014438-01A1
Application #
7142350
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Mcallister, Kimberly A
Project Start
2006-08-01
Project End
2008-06-30
Budget Start
2006-08-01
Budget End
2007-06-30
Support Year
1
Fiscal Year
2006
Total Cost
$191,868
Indirect Cost
Name
Wadsworth Center
Department
Type
DUNS #
153695478
City
Menands
State
NY
Country
United States
Zip Code
12204
Hwa Yun, Byeong; Guo, Jingshu; Bellamri, Medjda et al. (2018) DNA adducts: Formation, biological effects, and new biospecimens for mass spectrometric measurements in humans. Mass Spectrom Rev :
Guo, Jingshu; Turesky, Robert J (2016) Human Biomonitoring of DNA Adducts by Ion Trap Multistage Mass Spectrometry. Curr Protoc Nucleic Acid Chem 66:7.24.1-7.24.25
Turesky, Robert J; Le Marchand, Loic (2011) Metabolism and biomarkers of heterocyclic aromatic amines in molecular epidemiology studies: lessons learned from aromatic amines. Chem Res Toxicol 24:1169-214
Bessette, Erin E; Spivack, Simon D; Goodenough, Angela K et al. (2010) Identification of carcinogen DNA adducts in human saliva by linear quadrupole ion trap/multistage tandem mass spectrometry. Chem Res Toxicol 23:1234-44
Bessette, Erin E; Goodenough, Angela K; Langouët, Sophie et al. (2009) Screening for DNA adducts by data-dependent constant neutral loss-triple stage mass spectrometry with a linear quadrupole ion trap mass spectrometer. Anal Chem 81:809-19
Spink, Barbara C; Bennett, James A; Pentecost, Brian T et al. (2009) Long-term estrogen exposure promotes carcinogen bioactivation, induces persistent changes in gene expression, and enhances the tumorigenicity of MCF-7 human breast cancer cells. Toxicol Appl Pharmacol 240:355-66
Turesky, Robert J; Bendaly, Jean; Yasa, Isil et al. (2009) The impact of NAT2 acetylator genotype on mutagenesis and DNA adducts from 2-amino-9H-pyrido[2,3-b]indole. Chem Res Toxicol 22:726-33
Goodenough, Angela K; Schut, Herman A J; Turesky, Robert J (2007) Novel LC-ESI/MS/MS(n) method for the characterization and quantification of 2'-deoxyguanosine adducts of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by 2-D linear quadrupole ion trap mass spectrometry. Chem Res Toxicol 20:263-76