Effects of Pod E-Cigarettes on Respiratory Mucosal Immune Defenses

项目总结/摘要 电子烟是一个重大的公共卫生问题，超过700万成年人和500多万青年 报告目前的电子烟使用情况最近爆发的电子烟和电子烟产品相关肺损伤 EVALI进一步强调了研究电子烟使用影响的必要性。Pod电子烟，如Juul， 自2016年发布以来，它迅速受到欢迎，现在占电子烟市场的70%以上。 然而，人们对它们对人体呼吸系统的影响知之甚少。具体而言，无论和 它们如何影响呼吸道粘膜免疫防御，以及这些反应与 上一代电子烟（vape笔，坦克，盒子mods），代表了一个关键的知识差距， 这个应用程序的重点。我们将使用紧密联系的机制，在人体内和体外研究，以确定 豆荚电子烟如何影响呼吸道巨噬细胞，一种关键的呼吸道免疫细胞类型。初步数据 来自我们实验室和其他研究小组的研究表明，上一代电子烟及其成分 （调味化学品、丙二醇、植物甘油）有可能损害呼吸道粘膜 免疫防御，包括吞噬作用，纤毛跳动，氧化爆发，糖酵解，线粒体呼吸， 和免疫相关基因表达。由于JUUL独特的化学成分， 参数和抽吸地形，我们预计在JUUL用户中观察到额外或不同的影响， 与上一代电子烟使用者相比。我们将在两个具体目标中测试这一假设： 确定pod电子烟装置的用户是否在中央气道中具有独特的先天免疫反应， SA 2将研究雾化荚电子液体及其组分对离体巨噬细胞功能的影响。 为了达到SA 1，从电子烟使用者（前一代和pod队列）诱导痰液样本， 吸烟者和非吸烟者的免疫细胞组成，吞噬功能， 线粒体能量、形态学和免疫组织化学染色。还将对诱导痰进行分析， 呼吸道疾病的生物标志物，包括炎症、急性肺损伤和血管损伤。为了达到SA 2， 从支气管镜检查获得的原代人肺巨噬细胞将粘附到transwell上， 在气液界面处暴露于JUUL豆荚（烟草和薄荷味）、尼古丁盐和苯甲酸的气溶胶 酸单独使用一种新的体外暴露系统。然后对细胞进行细胞毒性、炎性 反应，吞噬功能，和生物能量的能力，以确定每个组成部分的e- 在这些端点上的液体。从这些目标中获得的数据将被整合， 对pod电子烟装置对呼吸免疫功能障碍的影响的机理洞察， 解决与Juul等pod电子烟的潜在健康影响相关的关键知识差距。

PROJECT SUMMARY/ABSTRACT E-cigarettes are a significant public health concern, with over 7 million adults and more than 5 million youth reporting current e-cigarette use. The recent outbreak of e-cigarette and vaping product associated lung injury (EVALI) further highlights the need for research on the effects of e-cigarette use. Pod e-cigarettes, such as JUUL, have rapidly gained popularity since their release in 2016 and now represent over 70% of the e-cigarette market. However, relatively little is known about their effects on the human respiratory system. Specifically, whether and how they affect respiratory mucosal immune defenses, and how these responses compare to those elicited by previous generation e-cigarettes (vape pens, tanks, box mods), represents a critical knowledge gap that will be the focus of this application. We will use tightly linked mechanistic human in vivo and in vitro studies to determine how pod e-cigarettes affect respiratory macrophages, a critical respiratory immune cell type. Preliminary data from our lab and other groups has demonstrated that previous generation e-cigarettes and their components (flavoring chemicals, propylene glycol, vegetable glycerin) have the potential to impair respiratory mucosal immune defenses, including phagocytosis, ciliary beating, oxidative burst, glycolysis, mitochondrial respiration, and immune-related gene expression. Because of JUUL’s unique chemical composition, aerosolization parameters, and puff topography, we anticipate observing additional or differential effects in JUUL users in comparison with previous generation e-cigarette users. We will test this hypothesis in two specific aims: SA1 will determine if users of pod e-cigarette devices have unique innate immune responses in the central airways and SA2 will investigate the effects of aerosolized pod e-liquid and its components on macrophage function ex vivo. To achieve SA1, induced sputum samples from e-cigarette users (both previous generation and pod cohorts), smokers, and nonsmokers will be analyzed for differences in immune cell composition, phagocytic function, mitochondrial energy, morphology, and immunohistochemical staining. Induced sputum will also be analyzed for biomarkers of airway disease, including inflammation, acute lung injury, and vascular damage. To achieve SA2, primary human pulmonary macrophages obtained from bronchoscopies will be adhered to transwells and exposed at air-liquid interface to aerosols from JUUL pods (tobacco & mint flavor), nicotine salts, and benzoic acid alone using a novel in vitro exposure system. The cells will then be assayed for cytotoxicity, inflammatory response, phagocytic function, and bioenergetic capacity to determine the effects of each component of the e- liquid on these endpoints. Data derived from these aims will be integrated to provide highly translational, mechanistic insight into the effects of pod e-cigarette devices on respiratory immune dysfunction, which will address a critical knowledge gap related to the potential health effects of pod e-cigarettes such as JUUL.