Development of biomarkers of exposure and effects for electronic cigarette vs. combustible cigarette use

虽然电子烟（e-cig）蒸汽中有毒物质和致癌物质的含量降低可能意味着健康水平降低 但是，风险并不等于没有风险。事实上，暴露于电子烟蒸汽的许多相同成分， 各种浓度，已与各种呼吸，心血管和免疫相关的 （炎性）疾病和癌症。到目前为止，电子烟的长期健康后果主要是 未知理由：电子烟蒸汽和香烟中存在的许多有害化学物质的生物效应 吸烟归因于它们诱导导致转录失调遗传/表观遗传变化的能力 与疾病相关的基因。方法：我们将进行多组学分析，以检测遗传/表观遗传 与口腔细胞和血液白细胞中关键功能基因转录失调相关的改变 健康成人vapers和吸烟者与对照组（非吸烟者）相比（N=45/组，匹配 年龄、性别和种族）。目标1：应用RNA测序和基因本体分析，“我们将首先筛选 与对照组相比，vapers和吸烟者的口腔和血细胞中疾病相关基因的失调“。 在确定了每组中失调的基因后，我们将采用有针对性的下一代测序技术， “检测Aim 2中的遗传变化以及Aim 3中失调基因内的表观遗传修饰”。 来自目标2和3的数据的整合分析将鉴定具有可检测的遗传/表观遗传变化的基因座， 以最高的灵敏度和特异性将电子烟使用者和吸烟者与对照组以及彼此区分开来。 与细胞中疾病相关基因异常表达相关的可检测遗传/表观遗传变化 从vapers与吸烟者可以作为暴露的新生物标志物和vaping与吸烟的影响。这些 生物标志物将有助于了解使用电子烟与吸烟的健康风险或潜在益处。作为次要目标， 我们将进行计算建模“，以找到所识别的生物标志物与受试者的 烟草产品使用频率和模式以及产品特性”。这将说明 电子烟/吸烟剂量和产品特性，包括电子烟装置特征（例如，模型和电源）和e- 液体含量（尼古丁和香料），以及香烟品牌、类型和化学成分（例如，焦油），在 电子烟使用与吸烟的生物效应。对RFA的响应：该提案将最大限度地 使用我们最近完成的NIDCR资助项目中的现有生物标本，其研究对象是 通过与USC-TCORS合作招募，由FDA-CTP赞助。这是一个独特 从加州的代表性人群中收集生物标本。中没有公开可用的存储库 美国提供了该项目所需的类似样品。标本目前保存在PI的实验室， 并且易于使用。影响：本项目的研究结果可推广到更广泛的人群， 美国和世界各地的电子烟使用者和吸烟者，因此可以立即用于科学界 和监管机构，并与公众的健康有很大的关系。

While the reduced levels of toxicants and carcinogens in electronic cigarette (e-cig) vapor may imply lower health risk, it cannot, however, equate to no risk. In fact, exposure to many of the same constituents of e-cig vapor, at various concentrations, has been associated with a variety of respiratory-, cardiovascular-, and immune-related (inflammatory) diseases, and cancer. To date, the long-term health consequences of vaping are largely unknown. Rationale: The biological effects of many harmful chemicals present in both e-cig vapor and cigarette smoke are ascribed to their ability to induce genetic/epigenetic changes that cause transcriptional deregulation of disease-related genes. Approach: We will perform a multi-omics analysis to detect genetic/epigenetic alterations associated with transcriptional deregulation of key functional genes in oral cells and blood leukocytes of healthy adult vapers and smokers in comparison to controls (nonsmokers non-vapers) (N=45/group, matched for age, sex, and race). Aim 1: Applying RNA-sequencing and gene ontology analysis, “we will first screen for deregulation of disease-related genes in oral- and blood cells of vapers and smokers as compared to controls”. Having identified the deregulated genes in each group, we will then employ targeted next-generation sequencing “to detect genetic changes in Aim 2, and epigenetic modifications in Aim 3 within the deregulated genes”. Integration analysis of data from Aims 2 and 3 will identify loci with detectable genetic/epigenetic changes that differentiate, with the highest sensitivity and specificity, vapers and smokers from controls and from one another. Detectable genetic/epigenetic changes associated with aberrant expression of disease-related genes in cells from vapers vs. smokers can serve as novel biomarkers of exposure and effects for vaping vs. smoking. These biomarkers will help inform the health risks or potential benefits of e-cig use vs. smoking. As a secondary goal, we will perform computational modeling “to find correlations between the identified biomarkers and subjects’ tobacco product use frequency and patterns and product characteristics”. This will inform the impact of vaping/smoking dose and product characteristics, including e-cig device features (e.g., model & power) and e- liquid content (nicotine and flavor), and cigarette brand, type, and chemical constituents (e.g., tar), on the biological effects of e-cig use vs. cigarette smoking. Responsiveness to RFA: This proposal will maximize the use of existing biospecimens from our recently completed NIDCR-funded project whose study subjects were recruited through collaboration with USC-TCORS, which is sponsored by the FDA-CTP. This is a unique collection of biospecimens from a representative population in California. No publicly available repository in the US offers similar specimens required for this project. The specimens are currently stored in the PI’s laboratory, and readily available for use. Impact: The findings of this project may generalize to the broader population of vapers and smokers in the US and across the world, thus being of immediate use for the scientific community and regulatory authorities, and of great relevance to the public’s health.