Exosomal epigenetic biomarkers associated with flavored electronic cigarette use in adults

项目总结 标题：与成人使用加味电子烟相关的外体表观遗传生物标记物 该项目涉及RFA-OD-19-021：1中定义的两个科学兴趣领域)利用血液和 用于分析烟草暴露生物标志物的尿样，目前其他国家尚未测量到 研究；2)检查与接触铅有关的外体表观遗传生物标记的差异 烟草产品。电子烟，一种模拟吸烟感觉的手持电子设备，获得了 过去几年全球流行，特别是在年轻人中，很大程度上是由于各种电子液体 味道。然而，使用电子烟的健康风险，特别是长期的健康风险尚不清楚。电子液体 通常含有丙二醇和/或植物甘油以及尼古丁和调味品。我们的 不同调味剂对肺上皮细胞和单核细胞影响的实验研究 不含尼古丁的电子烟香精表现出不同的细胞毒性。我们的初步数据显示， 在外体表观遗传生物标志物(微小RNA和长非编码RNA)方面存在显著差异 在非吸烟者和电子烟使用者之间使用人体血浆样本。然而，电子液体的味道是如何 (调味剂)导致电子烟启动期间外体表观遗传生物标记物的变化， 暴露，戒断在很大程度上是未知的。考虑到microoRNAs和MicroRNAs的关键调控功能 长的非编码RNA及其与疾病的关联，评估它们的变化和 香料电子烟的启动、暴露和戒烟过程中的相关生物途径 了解与使用电子烟相关的健康风险。拟议研究的首要目标是 识别与外体表观遗传相关的生物标记物(包括微小RNA和长的非编码RNA) 加了调味的电子烟。我们已经要求了两个纵向尿样(路径波1研究 2013至2014年和2014至2015年进行的第二波研究)和横断面血液 烟草与健康人口评估(PATH)研究样本(PATH波1研究) 生物仓库。样品可以从PATH根据生物素可用性报告(BAR)获得 ID：2019-005-BAR-2)从Path Study Biospecimen Access Program收到。我们将把这些链接起来 通过唯一的主题ID使用路径调查数据进行生物检疫。为了实现我们的目标，我们将检查这两个 血和尿外体表观遗传生物标记物(microRNA和长非编码RNA)及其相关 与加味电子烟使用有关的生物途径(如水果味)(目标1)，以及内部- 外体表观遗传生物标记物和相关生物通路在e-DNA损伤过程中的变化 开始吸烟和戒烟(目标2)。此外，根据确定的关键miRNAs和LncRNAs 对比目标1和目标2，我们将进行创新实验。我们将曝光这些miRNA并 对原代人支气管上皮细胞(HBEC)和小气道上皮细胞(SAEC)的非 吸烟者成人以确定他们的毒性/炎症反应，以供监管科学使用。的结果 监管科学：通过鉴定香料电子烟的健康风险 路径波1和波2中与电子烟暴露相关的microRNA和长非编码RNA 血液和尿液样本将提供有关电子烟调味品相对毒性的关键信息， 将提供证据支持FDA的监管努力。

PROJECT SUMMARY Title: Exosomal epigenetic biomarkers associated with flavored electronic cigarette use in adults This project addresses two scientific interest areas as defined in RFA-OD-19-021: 1) Utilize both blood and urine specimens to analyze biomarkers of tobacco exposure not currently measured by other national studies; 2) Examine differences in exosomal epigenetic biomarkers associated with exposure to tobacco products. E-cigarette, a handheld electronic device simulating the feeling of tobacco smoking, gains global popularity during the past few years especially in youth and young adults, largely due to various e-liquid flavors. However, the health risks of e-cigarette use, especially long-term health risks, are unclear. E-liquid usually contains propylene glycol and/or vegetable glycerin as well as nicotine and flavoring chemicals. Our previous experimental studies on lung epithelial cells and monocytes exposed to different flavoring chemicals or e-cigarette flavors without nicotine have shown different cytotoxicity. Our preliminary data showed that there were significant differences in exosomal epigenetic biomarkers (microRNAs and long non-coding RNAs) between non-users and e-cigarette users using human plasma samples. However, how e-liquid flavors (flavoring chemicals) lead to the changes in exosomal epigenetic biomarkers during the e-cigarette initiation, exposure, and cessation is largely unknown. Considering the crucial regulatory functions of micoRNAs and long non-coding RNAs and their association with diseases, it is critical to evaluate their changes and associated biological pathways during the flavored e-cigarette initiation, exposure and cessation to help understand health risks associated with e-cigarette use. The overarching goal of the proposed study is to identify exosomal epigenetic biomarkers (including microRNAs and long non-coding RNAs) associated with flavored e-cigarettes. We have requested both longitudinal urine specimens (PATH Wave 1 study conducted from 2013 to 2014 and Wave 2 study conducted from 2014 to 2015) and cross-sectional blood specimen (PATH Wave 1 study) from Population Assessment of Tobacco and Health (PATH) Study biorepositories. The samples are available from PATH based upon the Biospecimen Availability Report (BAR ID: 2019-005-BAR-2) received from PATH Study Biospecimen Access Program. We will link these biospecimens with PATH survey data through unique subject IDs. To achieve our goal, we will examine both blood and urinary exosomal epigenetic biomarkers (microRNAs and long non-coding RNAs) and associated biological pathways related to flavored e-cigarettes use (such as fruit flavor) (Aim 1), as well as the within- subject alterations in exosomal epigenetic biomarkers and associated biological pathways during the e- cigarette initiation and cessation (Aim 2). In addition, based on the identified key miRNAs and lncRNAs from aim 1 and aim 2 comparisons, we will conduct innovative experiments. We will expose those miRNAs and lncRNAs to primary human bronchial epithelial cells (HBEC) and small airway epithelial cells (SAEC) from non- smoker adults to determine their toxicity/inflammatory response for regulatory science. Outcomes for Regulatory Science: Assessment of health risks of the flavored e-cigarettes through identification of microRNAs and long non-coding RNAs associated with e-cigarette exposure from PATH Wave 1 and Wave 2 blood and urine specimens will provide crucial information on relative toxicity of e-cigarette flavorings, which would provide evidence to support the FDA’s regulatory efforts.