Integrated risk assessment and molecular characterization of pulmonary response to e-cigarette exposure

项目摘要 电子烟(e-cig)气溶胶/蒸气是电子烟液体(丙烯)原始成分的复杂混合物 甘醇(PG)、植物甘油(VG)、尼古丁、水和调味剂)以及其他成分(如 如醛、金属、纳米颗粒和一些未知化合物) 在氧气存在的情况下，暴露在电热金属丝中的原始部件。新兴 研究开始挑战人们对电子烟“相对安全”的看法。最近的研究表明电子烟 气雾剂/蒸汽引发炎症反应和氧化应激(使人想起香烟烟雾，如 氧化应激和炎症是与毒性和炎症相关的第一批临床定义事件 吸烟引起的疾病)，然而，其潜在的分子机制的细节仍不清楚。 此外，特定的电子液体成分在调解它们方面的贡献以及 人们对电子烟加热过程中产生的电子液体成分的氧化产物知之甚少。 将氧化应激和炎症作为生物反应预测的中间指标 组织功能障碍导致疾病的发生我们将评估电子液体主要成分的潜在危害 (PG、VG、尼古丁和调味品)。我们假设电子液体的个别成分会产生不同的 细胞和组织早期氧化/硝化损伤的特征。我们的目标是阐明这些签名 E-cig气溶胶诱导的可逆和不可逆的氧化修饰，并用它们来评估 不同电子液体成分的相对危险与计算流体力学- 用于比较呼吸剂量学的基于生理的药代动力学(CFD/PBPK)模型。多数 常用的细胞氧化还原状态(即谷胱甘肽含量)的测量方法很难深入了解 损伤诱导对于机械理解来说是至关重要的。定量氧化还原蛋白质组学的最新进展 PNNL使识别这些修改并确定其特定场地占有率成为可能 全蛋白质组的规模。巴特尔研发的研究级电子烟(REC)设备提供了独特的 从单个电子液体成分产生和表征气溶胶/蒸气的能力，无论是否有 使用加热线圈，允许主要电子液体成分以及潜在毒物的影响 在氧气存在的情况下对电子液体进行(盘管)加热的结果将被识别和量化。使用这些 新的使能技术，我们将定义哪些化合物或气溶胶颗粒组分是最有害的，以及如何 它们有助于吸入e-cig气雾剂和个别雾化e-液体成分对 肺部反应。我们将通过以下目标实现我们的目标：(1)表征气溶胶/水汽 中、高温下产生的电子烟(2)肺脏的分子特征 小鼠对e-cig气溶胶/蒸气和e-cig气溶胶/蒸气成分暴露的反应(3)预测区域性 (呼吸道)沉积和特定部位的组织剂量，以及使用CFD/PBPK模型对人体的转化。

PROJECT ABSTRACT E-cigarette (e-cig) aerosol/vapor is a complex mixture of the original components of the e-liquid (propylene glycol (PG), vegetable glycerol (VG), nicotine, water, and flavoring additives), and of other constituents (such as aldehydes, metals, nanoparticles, and some unknown compounds) produced by chemical transformation of the original components exposed to the electrically heated metal wire in the presence of oxygen. Emerging research is beginning to challenge the “relatively safe” perception of e-cigarettes. Recent studies suggest e-cig aerosol/vapor provokes an inflammatory response and oxidative stress (reminiscent of cigarette smoke where oxidative stress and inflammation are among the first clinically defined events associated with toxicity and disease due to cigarette smoking) however details of the underlying molecular mechanisms remain unclear. Furthermore the contributions that specific e-liquid constituents have in mediating them as well as the impact of oxidation products of e-liquid constituents generated by the e-cigarette heating process, are poorly understood. Focusing on oxidative stress and inflammation as intermediate measures of biological responses predictive of tissue dysfunction leading to disease initiation we will assess the hazard potential of e-liquid main components (PG, VG, nicotine, and flavorings). We hypothesize that individual components of e-liquid produce distinct signatures of early oxidative/nitrative damage in cells and tissue. Our objectives are to elucidate the signatures of reversible and irreversible oxidative modifications induced by e-cig aerosol and use them to evaluate the relative hazards of different e-liquid constituents in conjunction with computational fluid dynamic- physiologically-based pharmacokinetic (CFD/PBPK) models for comparative respiratory dosimetry. Most commonly used measures of cell redox state (i.e., GSH content) provide little insight into the types or sites of damage induced critical for a mechanistic understanding. Recent advances in quantitative redox proteomics at PNNL make it feasible to identify these modifications and determine their site-specific occupancies at a proteome-wide scale. The Research Grade E-cigarette (REC) device developed at Battelle provides a unique capability to generate and characterize aerosol/vapor from individual e-liquid components with or without the use of a heated coil, allowing effects of both the major e-liquid components as well as potential toxicants that result from (coil) heating of the e-liquid in the presence of oxygen to be identified and quantified. Using these novel enabling technologies, we will define what compounds or aerosol size fraction are most harmful and how they contribute to the effects of inhalation of e-cig aerosol and individual aerosolized e-liquid components on pulmonary response. We will accomplish our goal through the following aims: (1) Characterize aerosol/vapor of e-cigarette generated at moderate and high heating temperature (2) Molecularly characterize pulmonary response to e-cig aerosol/vapor and e-cig aerosol/vapor components exposure in mice (3) Predict regional (airway) deposition and site-specific tissue dose and translation to humans using CFD/PBPK models.