Identification of Free Radical Induced Biomarkers of Exposure to Electronic Cigarette Aerosol

暴露于电子烟气溶胶的自由基诱导生物标志物的鉴定

• 批准号: 9981091
• 负责人: Zachary T Bitzer
• 金额: $ 12.05万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981091
• 关键词: Address; Adoption; Aerosols; Animals; Antibodies; Area; Award; Biological; Biological Markers; C57BL/6 Mouse; Characteristics; Chemicals; Chronic Obstructive Airway Disease; DNA Adducts; Data; Detection; Development; Disease; Electron Spin Resonance Spectroscopy; Electronic cigarette; Ensure; Exposure to; Free Radicals; Freezing; Glycerol; Health; Inflammation; Lead; Long-Term Effects; Lung; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mentorship; Modeling; Mus; Nose; Oxidants; Oxides; Pathway interactions; Phase; Physiologic pulse; Propylene Glycols; Proteins; Research; Research Personnel; Rodent; Rodent Model; Safety; Science; Series; Serum; Solvents; Spin Trapping; Structure; Techniques; Time; Tissues; Tobacco; Tobacco smoke; Training; Translational Research; Work; Youth; adduct; assault; base; biomarker development; e-cigarette aerosols; electronic cigarette use; electronic liquid; environmental tobacco smoke exposure; experience; in vivo; insight; metabolomics; novel; oxidative damage; pyrroline; specific biomarkers; symposium; tobacco regulatory science; tool; toxicant; tumor progression; vaping

Project Summary/Abstract: Electronic cigarette (e-cig) usage is on the rise, particularly among youths; however, their potential for harm is not understood, complicating development of informed regulatory strategies. The lack of data on e-cig related harm is, in part, due to the lack of specific biomarkers for exposure to e-cig aerosols. We found that e-cig aerosol contains highly reactive free radicals that can cause oxidative damage to the user. Free radicals can damage numerous cellular pathways possibly contributing to the progression of cancers and other diseases. Detection of e-cig free radicals can be accomplished by trapping with spin traps (e.g., DMPO) and analysis by electron paramagnetic resonance (EPR) spectroscopy. Preliminary research with EPR shows that free radicals produced in the e-cig aerosol by e-liquid solvents, propylene glycol (PG) and glycerin (GLY), common to all e-cigs, display unique structural characteristics. Our objective is to identify these free radical structures and utilize their unique structure to develop an e-cig specific biomarker of exposure. The specific aims of the proposed research are: (Aim 1) To determine the structures of the free radicals produced by PG and GLY in e-cigs; (Aim 2) To determine the primary targets of and adducts formed from free radical assault in the tissue of e-cig exposures in rodent models and possible metabolites formed from the these radical adducts in the serum of e-cig exposed rodent. This project represents an important research direction where a chemical/biological approach can inform tobacco regulatory science. As such, an important aspect of this application it to extend my background in areas relevant to translational science in addition to providing specific training in new biomarker-relevant research areas including metabolomics and free radical structural identification. To this end, my training will occur through a series of courses, relevant mentorship, and practical experience, each geared to ensure my transition to an independent researcher in the fields of biomarker development and regulatory science. Coursework, mentorship, conference participation, and practical training/experience will be completed during the K99 phase. During this phase, Aim 1 of the research plan will be completed and Aim 2 will be initiated (for completion during the R00 phase of the award). To accomplish the research aims, advanced pulsed EPR and mass spectroscopy approaches will be utilized for radicals produced by PG and GLY in e-cig aerosols. In a mouse exposure model, free radical exposure and targets of attack in the lung will be determined using a novel in vivo DMPO/anti-DMPO antibody approach. This will allow for the identification of specific protein and DNA adducts by TOF-MS. A post-DMPO exposure study will consist of an untargeted pairwise metabolomics approach to look at changes in metabolite profiles before and after e-cig exposure. By leveraging the unique structures of e-cig produced free radicals and their targets of attack in the lung, biomarkers of exposure specific to e-cig aerosols can be identified and used to develop regulatory strategies aimed at reducing harm from e-cig exposure.

项目摘要/摘要：电子烟（e-cig）的使用量正在上升，尤其是在年轻人中； 然而，它们的潜在危害尚不为人所知，这使得制定明智的监管策略变得复杂化。 缺乏有关电子烟相关危害的数据，部分原因是缺乏接触电子烟的特定生物标志物 气溶胶。我们发现电子烟气溶胶含有高活性自由基，可导致氧化损伤 用户。自由基会损害许多细胞通路，可能导致疾病的进展 癌症和其他疾病。电子烟自由基的检测可以通过自旋陷阱捕获来完成 （例如 DMPO）和电子顺磁共振 (EPR) 光谱分析。初步研究与 EPR 显示，电子烟气溶胶中的电子液体溶剂、丙二醇 (PG) 和 所有电子烟都含有甘油 (GLY)，它具有独特的结构特征。我们的目标是识别这些 自由基结构并利用其独特的结构开发电子烟特定的暴露生物标志物。这 拟议研究的具体目标是：（目标 1）确定产生的自由基的结构 电子烟中的 PG 和 GLY； （目标 2）确定自由基的主要目标和由自由基形成的加合物 啮齿动物模型中电子烟暴露对组织的攻击以及这些自由基形成的可能代谢物 暴露于电子烟的啮齿动物血清中的加合物。该项目代表了一个重要的研究方向 化学/生物方法可以为烟草监管科学提供信息。因此，这方面的一个重要方面是 除了提供具体的知识之外，应用它来扩展我在转化科学相关领域的背景 新生物标志物相关研究领域的培训，包括代谢组学和自由基结构 鉴别。为此，我的培训将通过一系列课程、相关指导和实践进行 经验，每一项都旨在确保我过渡到生物标志物领域的独立研究员 发展和监管科学。课程作业、指导、会议参与和实践 培训/经验将在K99阶段完成。在此阶段，研究计划的目标 1 将 完成并启动目标 2（在奖励的 R00 阶段完成）。为了实现 研究目标，先进的脉冲 EPR 和质谱方法将用于产生自由基 电子烟气溶胶中的 PG 和 GLY。在小鼠暴露模型中，自由基暴露和攻击目标 将使用新型体内 DMPO/抗 DMPO 抗体方法来确定肺部。这将允许 通过 TOF-MS 鉴定特定蛋白质和 DNA 加合物。 DMPO 后暴露研究将包括 非靶向配对代谢组学方法观察电子烟前后代谢物谱的变化 接触。通过利用电子烟的独特结构产生自由基及其攻击目标 在肺部，可以识别电子烟气溶胶特异性暴露的生物标志物并用于制定监管规定 旨在减少电子烟暴露危害的策略。