Exosomal epigenetic biomarkers associated with flavored electronic cigarette use in adults

项目摘要 标题：与成人使用调味电子烟相关的外泌体表观遗传生物标志物 本项目涉及RFA-OD-19-021中定义的两个科学兴趣领域：1）利用血液和 尿液样本分析烟草暴露的生物标志物，目前没有其他国家测量 研究; 2）检查与暴露于以下物质相关的外来体表观遗传生物标志物的差异： 烟草制品.电子烟，一种模拟吸烟感觉的手持电子设备， 在过去的几年里，特别是在年轻人和年轻人中，全球流行，主要是由于各种电子液体 口味.然而，使用电子烟的健康风险，特别是长期健康风险尚不清楚。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化学品。我们 先前对暴露于不同调味化学品的肺上皮细胞和单核细胞的实验研究 或不含尼古丁的电子烟香料显示出不同的细胞毒性。初步数据显示， 外泌体表观遗传生物标志物（microRNA和长链非编码RNA） 使用人类血浆样本的非吸烟者和电子烟使用者之间的差异。然而，如何电子液体口味 （调味化学品）导致电子烟开始期间外泌体表观遗传生物标志物的变化， 暴露和停止在很大程度上是未知的。考虑到微小RNA的重要调控功能， 长链非编码RNA及其与疾病的关系，评估它们的变化和 相关的生物学途径在调味电子烟的启动，暴露和停止，以帮助 了解与电子烟使用相关的健康风险。拟议研究的总体目标是 鉴定相关外泌体表观遗传生物标志物（包括微小RNA和长非编码RNA） 有香味的电子烟我们已经要求了两个纵向尿液样本（PATH Wave 1研究 2013年至2014年进行的第2波研究）和横断面血液 烟草与健康人口评估研究（PATH Wave 1 Study） 生物储存库。根据生物样本可用性报告（BAR），可从PATH获得样本 ID：2019-005-BAR-2），接收自PATH研究生物样本访问项目。我们将把这些 通过唯一的受试者ID将PATH调查数据与生物标本进行比较。为了实现我们的目标，我们将研究 血液和尿液外泌体表观遗传生物标志物（microRNA和长链非编码RNA）和相关的 与调味电子烟使用相关的生物学途径（如水果味）（目标1），以及 受试者在e- 吸烟开始和停止（目标2）。此外，基于从细胞中鉴定的关键miRNAs和lncRNAs， 目标1和目标2的比较，我们将进行创新实验。我们将暴露这些miRNAs， lncRNA对来自非人支气管上皮细胞（HBEC）和小气道上皮细胞（SAEC）的作用 吸烟成年人，以确定其毒性/炎症反应的监管科学。成果 监管科学：通过识别调味电子烟的健康风险评估 来自PATH Wave 1和Wave 2的与电子烟暴露相关的microRNA和长链非编码RNA 血液和尿液样本将提供有关电子烟调味剂相对毒性的重要信息， 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.