Differentiating the biological effects of vaping from smoking by analyzing the methylome and transcriptome

对NOT-OD-22-022的响应性：这项提案将分离电子烟的生物效应 (e-cig)通过测量与吸烟风险相关的表观遗传和转录变化来使用(‘电子烟’) 健康成人尿布和/或吸烟者与对照组(非吸烟者)细胞和组织中的疾病 两种产品中的任何一种)。此外，它还将确定这些分子变化是否受 蒸发/吸烟的强度和持续时间以及所使用的烟草产品的特性(S)，包括电子烟 设备特征和电子液体成分，以及香烟品牌、类型和化学成分。通过阐明 蒸发与吸烟生物效应的分子机制，我们将开发新的生物标记物 暴露和影响，这将对评估健康风险或潜在的好处具有重要作用 与吸烟相关的蒸发。理由：成年电子烟使用者可能有吸烟史或共同使用史 电子烟和可燃香烟(即双重使用者)。为研究使用电子烟对成年人的生物影响， 当务之急是梳理出蒸发的后果，同时考虑“过去”或“过去”的混乱影响 “现在”吸烟。前提：电子烟蒸气和香烟烟雾中存在的许多有毒物质和致癌物质 它们通过表观遗传效应产生的生物学效应，例如异常的DNA甲基化和/或转录 可能导致疾病相关基因失调的改变。项目大纲：利用银行的杠杆作用 来自我们最近完成的NIDCR和TRDRP资助的研究的样本，我们将区分生物 通过分析口腔中的整个甲基组和转录组，电子烟的使用与吸烟的后果- 以及健康成人专用尿布、双重使用者、专用吸烟者和对照组的血细胞。我们 将使用一种新的方法，结合对整个甲基组(AIM 1)和转录组(AIM)的初级分析 2)和基于蒸发/吸烟量和烟草产品建立的各种模型的序贯敏感性分析 特点(目标3)。对目标1和目标2的数据进行综合分析将确定甲基组和 调节疾病特异性基因的转录组变化，从而识别暴露和死亡的新生物标志物 对蒸发、两用和吸烟的影响。我们将在患者群体中验证已识别的生物标志物 经过高度精心策划和质量控制的来自公共来源的“组学”数据集。这将验证 确定的生物标志物，用于评估专用电子烟使用者、双重使用者和专用吸烟者的疾病风险。 AIM 3中详细的计算建模和灵敏度分析将确定 蒸发/吸烟剂量和产品特性对尿布中甲基组和转录组变化的影响 双重使用者和吸烟者。这些数据将为FDA对烟草产品的监管提供信息，以保护公众健康。 创新：我们将研究电子烟气雾剂和香烟烟雾对尿布细胞和组织的影响 和/或吸烟者使用全基因组甲基组和转录组测序和综合生物信息学 分析和计算模型，考虑蒸发/吸烟剂量和产品特性。

Responsiveness to NOT-OD-22-022: This proposal will segregate the biological effects of electronic cigarette (e-cig) use (‘vaping’) from smoking by measuring the epigenetic and transcriptomic changes linked to risk of disease in cells and tissues of healthy adult vapers and/or cigarette smokers as compared to controls (non-users of either product). Furthermore, it will determine whether these molecular alterations are modulated by the intensity and duration of vaping/smoking and the characteristics of tobacco product(s) used, including e-cig device features and e-liquid ingredients, and cigarette brand, type, and chemical constituents. By elucidating the molecular mechanisms underlying the biological effects of vaping vs. smoking, we will develop novel biomarkers of exposure and effects, which will have significant utility for assessing the health risks or potential benefits of vaping relative to smoking. Rationale: Adult e-cig users are likely to have a prior history of smoking or co-use e-cigs and combustible cigarettes (i.e., dual users). To investigate the biological effects of e-cig use in adults, it is imperative to tease out the consequences of vaping, while accounting for the confounding effects of ‘past’ or ‘present’ smoking. Premise: Many toxicants and carcinogens present in e-cig vapor and cigarette smoke exert their biological effects through epigenetic effects, such as aberrant DNA methylation, and/or transcriptomic alterations that can lead to dysregulation of disease-related genes. Project outline: Leveraging the banked specimens from our recently completed NIDCR- and TRDRP-funded studies, we will differentiate the biological consequences of e-cig use from those of smoking by analyzing the whole methylome and transcriptome in oral- and blood cells of healthy adult ‘exclusive’ vapers, dual users, ‘exclusive’ cigarette smokers, and controls. We will use a novel approach, combining primary analysis of the whole methylome (Aim 1) and transcriptome (Aim 2) and ordinal sensitivity analysis of various models built on vaping/smoking dose and tobacco product characteristics (Aim 3). Integrative analysis of data from Aims 1 and 2 will determine the methylome and transcriptome changes that regulate disease-specific genes, thus identifying novel biomarkers of exposure and effects for vaping, dual use, and smoking. We will validate the identified biomarkers in patient populations using highly curated and quality-controlled ‘omics’ datasets processed from public sources. This will verify the utility of the identified biomarkers for assessing disease risk in exclusive’ e-cig users, dual users, and ‘exclusive’ smokers. The detailed computational modeling and sensitivity analysis in Aim 3 will determine the impact of vaping/smoking dose and product characteristics on the alterations of the methylome & transcriptome in vapers, dual users, and smokers. These data will inform the FDA’s regulation of tobacco products to protect public health. Innovation: We will investigate the effects of e-cig aerosol and cigarette smoke on cells and tissues of vapers and/or smokers using genome-wide sequencing of the methylome & transcriptome and integrative bioinformatic analysis & computational modeling, accounting for vaping/smoking dose and product characteristics.