Project 4 - Oral Cell DNA Adducts and Urinary Biomarkers to Investigate Ethnic/Racial Differences in Lung Cancer Susceptibility

项目4.口腔细胞DNA加合物和尿液生物标记物，以调查种族/种族差异 肺癌易感性。 摘要 我们正在进行的研究表明，致癌物质和有毒物质的剂量占到了一些种族 在多种族队列中观察到的吸烟者患肺癌风险的差异。致癌物质和 毒物剂量只是吸烟相关肺癌风险范例的第一部分。DNA加合物形成 是下一个关键步骤，因为DNA加合物会导致在吸烟者肺部发现的多种突变 这会导致编码错误、遗传不稳定和癌症。因此，在这项研究中，我们将比较DNA加合物 来自三个不同风险人群的吸烟者的形成：夏威夷原住民(最高风险 风险)、白人(风险较低)和日裔美国人(风险最低)。三百名受试者将由 临床和生物标志物核心。口腔细胞将作为肺细胞的替代品，用于DNA加合物 测量；将收集尿样以分析致癌物质和有毒代谢物。特定的 DNA加合物以及DNA加合物签名将从每个受试者的口腔细胞中获得。分析 以及之前从从不吸烟的人身上收集的尿样将检验这一假设 夏威夷原住民对肺癌的高易感性部分是由于内源性毒物的产生 丙烯醛和巴豆醛，可能是由于炎症和氧化损伤。因此，我们的具体目标是： 1.用高分辨质谱仪对100例口腔粘膜细胞中已知的DNA加合物进行定量 来自每个种族的吸烟者-夏威夷原住民、白人和日裔美国人。黄曲霉毒素DNA加合物 烟草特有的化合物、甲醛和丙烯醛将被量化。 2.分析每组100名吸烟者和100名不吸烟者的尿液中硫代尿酸的含量 丙烯醛和巴豆醛，F2-异前列腺素8-异前列腺素F-2α，氧化损伤的生物标记物，以及 前列腺素E2代谢物PGEM，炎症的生物标志物，以及总尼古丁当量和 NNAL总数(仅限吸烟者)。这些数据将提供与本地人感染高风险相关的关键信息 夏威夷人的肺癌，以及与特定目标的DNA加合物测量的关系1。 3.使用我们新开发的高分辨率质谱学、高通量DNA加合提取方法 筛选已知和未知的DNA加合物以识别全面的DNA修饰签名 源自吸烟。 A.使用有针对性的DNA内收切除法，我们将筛选多个DNA修饰 同时。在特定目标1中分析的加合物将被添加到一份清单中，包括 内源性，1，3-丁二烯衍生(与项目3合作)，醛衍生和 亚硝胺或烷化剂衍生的DNA加合物。与此同时，我们将应用我们的DNA内收切割术 方法以非靶向模式调查以前未知的DNA加合物的存在。 通过非靶向分析检测到的未知加合物将被包括在靶向列表中 DNA加合物最终获得一种敏感的方法来评估由 抽烟。 B.在特定目标3a中创建的DNA加合物列表将用于研究 特定目的分析的选定样本的DNA损伤特征1。 这项独特和创新的研究结果将提供与不同风险相关的关键信息 夏威夷原住民、白人和日裔美国人中的肺癌。我们期待着新一代 与吸烟有关的DNA加合物特征以及有关DNA加合物和尿代谢物的重要数据 已知的致癌物质和有毒物质。这些结果将为DNA加合物和DNA加合物之间的关系提供关键的检验。 某些尿代谢物与肺癌风险的关系，从而可能为肺癌预防提供新的见解 战略。

Project 4. Oral Cell DNA Adducts and Urinary Biomarkers to Investigate Ethnic/Racial Differences in Lung Cancer Susceptibility. ABSTRACT Our ongoing studies demonstrate that carcinogen and toxicant doses account for some of the ethnic differences in risk for lung cancer among smokers as observed in the Multiethnic Cohort. Carcinogen and toxicant dose are only the first part of the risk paradigm for smoking-related lung cancer. DNA adduct formation is the next critical step because DNA adducts lead to the multiple mutations that are found in smokers' lungs and that cause miscoding, genetic instability, and cancer. Therefore, in this study, we will compare DNA adduct formation in smokers from three ethnic groups with differing risks for lung cancer: Native Hawaiians (highest risk), Whites (lower risk), and Japanese Americans (lowest risk). Three-hundred subjects will be recruited by the Clinical and Biomarker Core. Oral cells, as a surrogate for lung cells, will be obtained for DNA adduct measurements; urine samples will be collected for analysis of carcinogen and toxicant metabolites. Specific DNA adducts as well as DNA adductomic signatures will be obtained from oral cells of each subject. Analysis of the urine samples as well as those previously collected from never smokers will test the hypothesis that the high lung cancer susceptibility of Native Hawaiians is partially due to endogenous generation of the toxicants acrolein and crotonaldehyde, possibly due to inflammation and oxidative damage. Thus, our specific aims are: 1. 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 our 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. a. Using a targeted DNA adductomic method we will 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 we will apply our DNA adductomic method 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 . b. 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. The results of this unique and innovative study will provide critical information relevant to the differing risks for lung cancer among Native Hawaiians, Whites, and Japanese Americans. We expect generation of new smoking-related DNA adduct signatures as well as important data on DNA adducts and urinary metabolites of known carcinogens and toxicants. The results will provide a critical test of the relationship of DNA adducts and certain urinary metabolites to lung cancer risk, thus possibly leading to new insights for lung cancer prevention strategies.