Inflammatory and Dysregulated Repair Responses to Inhaled Nicotine

总结 尼古丁是尼古丁输送系统[电子尼古丁输送系统（ENDS）]的主要成分，即 电子烟（Electronic Cigs）众所周知，尼古丁具有成瘾性，存在知识缺口 吸入尼古丁对肺部系统的影响我们的支持数据表明， 递送和暴露在人肺上皮细胞中引起氧化应激和炎症反应， 成纤维细胞和小鼠肺中。目前，没有关于电子烟生物效应的信息 在人类和小鼠模型中含有吸入的尼古丁。吸入尼古丁可能会导致 肺部疾病的发病机制，特别是通过肺部炎症、损伤和失调修复 应答我们假设，电子烟尼古丁影响毒性证明了氧化和炎症 在人类和小鼠模型中的反应，导致失调的修复和肺气肿反应。 提出了三个具体目标来检验这一假设： 目的1：吸入尼古丁诱导人类受试者的肺部和全身炎症介质 确定吸入尼古丁对电子烟使用者和非使用者的影响。这将通过 监测人体暴露的生物标志物（通过脂质组学检测炎症、外来体和脂质介质） 生物流体（唾液、呼出气冷凝液、血浆和尿液）沿着临床结果（肺功能 在前瞻性队列研究中（基线和随访）。沿着人类研究，我们计划进行 在体内和体外的机制研究。 目的2：吸入尼古丁通过其受体诱导肺部炎症和失调修复反应 在这里，我们将使用小鼠临床前模型进行机制研究。我们将确定电子烟是否 含有低浓度和高浓度尼古丁的组合物具有不同的促炎和异常修复作用 在体内通过α7烟碱乙酰胆碱受体（α 7 nAChR）依赖性机制。 目的3：尼古丁气雾剂诱导炎症和失调细胞修复的机制 响应 确定人类肺部对电子烟尼古丁蒸汽的炎症和失调细胞修复反应 上皮细胞和成纤维细胞使用最先进的报告模型（NF-κB荧光素酶）以及三维细胞 文化模式这将决定尼古丁如何影响细胞过程，如早期细胞衰老 和肌成纤维细胞分化，以及愈合/修复过程中的脂肪生成和肌生成途径。 这项研究的结果将提供对临床影响和机制的理解， 人类受试者尼古丁暴露后的炎症、衰老和失调修复反应 在体外原代肺细胞和小鼠体内模型中。

SUMMARY Nicotine is a major component of nicotine delivery systems [Electronic Nicotine Delivery Systems (ENDS)] i.e. electronic cigarettes (e-cigs). Nicotine is known to have the addictive properties, and a knowledge gap exists on how inhaled nicotine affects the pulmonary system. Our supporting data show that ENDS nicotine aerosol delivery and exposure cause oxidative stress and inflammatory responses in human lung epithelial cells, fibroblasts, and in mouse lungs. Currently, no information is available on the biological effects of e-cig containing inhaled nicotine in humans and in mouse models. Inhaled nicotine may contribute to the pathogenesis of lung diseases in particular via lung inflammation, injurious, and dysregulated repair responses. We hypothesize that e-cig nicotine influences toxicity as evidenced by oxidative and inflammatory responses in humans and in mouse models, leading to dysregulated repair and emphysematous responses. Three specific aims are proposed to test this hypothesis: Aim 1: Inhaled nicotine induces lung and systemic inflammatory mediators in human subjects Determine the impact of inhaled nicotine in users and non-users of e-cigarettes. This will be accomplished by monitoring biomarkers of exposure (inflammatory, exosomes and lipid mediators by lipidomics) in human biofluids (saliva, Exhaled Breath Condensate, plasma, and urine) along with clinical outcomes (lung function tests) in a prospective cohort study (baseline and follow-up). Along with human studies, we plan to conduct mechanistic studies in vivo and in vitro. Aim 2: Inhaled nicotine induces lung inflammatory and dysregulated repair responses via its receptor Here, we will use a mouse preclinical model for mechanistic studies. We will determine if e-cigarettes containing low and high nicotine concentrations have differential pro-inflammatory and abnormal repair effects in vivo via the α7 nicotinic acetylcholine receptor (α7nAChR) dependent mechanism. Aim 3: Mechanisms whereby nicotine aerosol induces inflammatory and dysregulated cellular repair responses Determine inflammatory and dysregulated cellular repair responses to e-cigarette nicotine vapor in human lung epithelial cells and fibroblasts using the state-of-the-art reporter models (NF-κB luciferase) as well as a 3-D cell culture model. This will determine how nicotine affects cellular processes, such as early cellular senescence and myofibroblast differentiation, as well as lipogenic and myogenic pathways in healing/repair process. The outcomes of this study will provide an understanding of the clinical impact and mechanisms of inflammatory, senescence, and dysregulated repair responses following nicotine exposure in human subjects and, in primary lung cells in vitro and mouse model in vivo.