High resolution mass spectrometric profile analysis of carcinogen-DNA adducts in oral cells of cigarette smokers and squamous cell carcinoma of the head and neck

头颈部鳞状细胞癌（HNSCC）是一种毁灭性的，经常毁容， 这种致命的疾病预计将在2020年影响美国超过53，000人，并导致超过10，000人死亡。 预防这种可怕的疾病至关重要。吸烟、饮酒与人乳头状瘤 病毒（HPV）是HNSCC的主要原因;只有吸烟和饮酒是 在这里考虑。吸烟和酒精饮料是多种DNA加合物的来源， 在致癌过程中。该提案将建立一种液相色谱-纳米电喷雾离子化- 12种口腔细胞DNA加合物的高分辨串联质谱分析 可能导致HNSCC的原因这是受到我们最近对口腔细胞中DNA加合物的分析的启发， 我们发现吸烟者的水平比不吸烟者高出20倍。这些令人兴奋 这些结果鼓励我们提出一个口腔细胞中重要致癌物衍生DNA加合物的图谱分析 根据以下具体目标： 1.开发LC-NSI-HRMS/MS图谱分析方法，用于定量12种重要和代表性的 致癌物和有毒物-人类口腔细胞和组织中的DNA加合物。加合物衍生自 香烟烟雾和酒精饮料中的各种致癌物质和DNA反应化合物。 2.将谱分析应用于来自当前健康个体的口腔细胞： 饮酒者或轻度饮酒者; B）100名不饮酒者或轻度饮酒者的吸烟者;以及c）100名 适度或重度饮酒的吸烟者。组a和B中加合物水平的比较 将鉴定吸烟增强的加合物，而B和c组的比较将鉴定 加合物，增强了吸烟和中度或重度饮酒的组合。 3.在50名受试者中测试6个月内口腔细胞DNA加合物谱分析的纵向稳定性 不饮酒或少量饮酒的吸烟者。 4. A）确定从75名患有HNSCC的吸烟者收集的口腔细胞中的DNA加合物谱，并与 在特定目标2中招募的200名无HNSCC的吸烟者中， 这是HNSCC发病率特征。 B）将来自该目的的A部分的口腔细胞DNA加合物谱与正常和非正常组织中的口腔细胞DNA加合物谱进行比较。 肿瘤，在来自A部分的60名接受手术的患者的亚组中，以确定口腔细胞DNA是否 加合物模式与组织中的模式一致。 我们的研究结果将有可能识别出易患HNSCC但无法戒烟的个体。 一旦确定，积极的生活方式和监测干预措施，如口腔检查，这些受试者 2-4每年可以启动10次，以预防或早期发现这种毁容且往往致命的癌症。

Squamous cell carcinoma of the head and neck (HNSCC) is a devastating, frequently disfiguring, and often fatal disease, expected to affect more than 53,000 people in the U.S. in 2020 and kill more than 10,000. Prevention of this terrible disease is critical. Cigarette smoking, alcohol consumption, and human papilloma virus (HPV) are well established major causes of HNSCC; only smoking and alcohol consumption are considered here. Cigarette smoking and alcoholic drinks are sources of multiple DNA adducts that are critical in the carcinogenic process. This proposal will establish a liquid chromatography-nanoelectrospray ionization- high resolution tandem mass spectrometry (LC-NSI-HRMS/MS) profile analysis of 12 oral cell DNA adducts that are likely causes of HNSCC. This was inspired by our recent analysis of DNA adducts in oral cells, in which we found levels more than 20 times higher in cigarette smokers than in non-smokers. These exciting results encouraged us to propose a profile analysis of important carcinogen-derived DNA adducts in oral cells according to the following specific aims: 1. Develop an LC-NSI-HRMS/MS profile analysis method for quantitation of 12 important and representative carcinogen and toxicant – DNA adducts in human oral cells and tissue. The adducts are derived from various carcinogens and DNA reactive compounds in cigarette smoke and alcoholic beverages. 2. Apply the profile analysis to oral cells from currently healthy individuals: a) 100 non-smokers who are non- drinkers or light drinkers; b) 100 cigarette smokers who are non-drinkers or light drinkers; and c) 100 cigarette smokers who are moderate or heavy drinkers. Comparisons of adduct levels in groups a and b will identify adducts enhanced by cigarette smoking while comparisons of groups b and c will identify adducts that are enhanced by the combination of smoking and moderate or heavy drinking. 3. Test the longitudinal stability of the oral cell DNA adduct profile analysis over a 6 month period in 50 smokers who are non-drinkers or light drinkers. 4. A) Determine the DNA adduct profile in oral cells collected from 75 smokers with HNSCC and compare to that in 200 smokers without HNSCC recruited in Specific Aim 2 with the goal of identifying an adduct profile that is characteristic of HNSCC incidence. B) Compare the oral cell DNA adduct profile from part A of this aim to that in tissue, both normal and tumor, in a subset of 60 patients from part A who undergo surgery, to determine whether oral cell DNA adduct patterns are consistent with those in tissue. Our results will potentially identify individuals who are susceptible to HNSCC but are unable to quit smoking. Once identified, aggressive lifestyle and monitoring interventions in these subjects such as oral examinations 2-4 times per year can be initiated for prevention or early detection of this disfiguring and often fatal cancer.