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，经常使人毁容，通常 一种致命的疾病，预计到2020年，美国将有超过5.3万人感染这种疾病，并导致超过1万人死亡。 预防这种可怕的疾病至关重要。吸烟、饮酒和人乳头状瘤 病毒(HPV)是公认的HNSCC的主要原因；只有吸烟和饮酒是 在这里考虑过。吸烟和酒精饮料是多种DNA加合物的来源，这些加合物是至关重要的 在致癌过程中。这项提议将建立一种液体色谱-纳米电喷雾电离- 12种口腔细胞DNA加合物的LC-NSI-HRMS/MS谱分析 这可能是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，在 我们发现吸烟者体内的这种物质水平是不吸烟者的20多倍。这些令人兴奋的东西 结果鼓励我们提出对口腔细胞中重要致癌物衍生的DNA加合物进行轮廓分析。 根据以下具体目标： 1.建立了LC-NSI-HRMS/MS图谱分析方法，用于12个重要和有代表性的成分的定量 人类口腔细胞和组织中的致癌物质和有毒DNA加合物。加合物是从 香烟烟雾和酒精饮料中的各种致癌物质和DNA活性化合物。 2.将轮廓分析应用于来自当前健康个人的口腔细胞：a)100名非吸烟者 饮酒者或饮酒者；b)100名吸烟者，但不饮酒者或饮酒者；以及c)100人 适度或重度饮酒的吸烟者。A组和B组加合物水平的比较 将确定吸烟增加的加合物，而b组和c组的比较将确定 吸烟和适度或大量饮酒相结合而增强的加合物。 3.检验口腔细胞DNA加合物图谱分析在6个月期间的纵向稳定性 不饮酒或少量饮酒的吸烟者。 4.a)测定75例HNSCC吸烟者口腔细胞的DNA加合物谱，并与 在特定目标2中招募的200名没有HNSCC的吸烟者中，目标是确定加合物分布 这是HNSCC发病的特点。 B)将本目标A部分的口腔细胞DNA加合物图谱与组织中的DNA加合物图谱进行比较，正常和 肿瘤，来自A部分的60名接受手术的患者的子集，以确定口腔细胞DNA 加合物模式与组织中的一致。 我们的结果可能会识别出易患HNSCC但无法戒烟的人。 一旦确定，积极的生活方式和对这些对象的监测干预，如口腔检查 每年可以启动2-4次，以预防或早期发现这种毁容的、往往是致命的癌症。

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.