Effects of Pod E-Cigarettes on Respiratory Mucosal Immune Defenses

项目摘要/摘要 电子烟是一个重大的公共卫生问题，有700多万成年人和500多万青年 报告当前电子烟的使用情况。最近爆发的电子烟和电子烟产品相关的肺损伤 (EVALI)进一步强调了研究电子烟使用的影响的必要性。豆荚电子烟，如Juul， 自2016年发布以来，电子烟迅速流行起来，目前占电子烟市场的70%以上。 然而，人们对它们对人类呼吸系统的影响知之甚少。具体地说，是否和 它们如何影响呼吸道粘膜免疫防御，以及这些反应与由 上一代电子烟(电子烟笔、罐、盒式MOD)代表着一个关键的知识鸿沟，将是 这个应用程序的重点。我们将使用紧密相连的人体体内和体外机械性研究来确定 豆荚电子烟如何影响呼吸巨噬细胞，这是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小组的研究表明，上一代电子烟及其组件 (调味品、丙二醇、蔬菜甘油)有可能损害呼吸道粘膜 免疫防御，包括吞噬、纤毛跳动、氧化爆发、糖酵解、线粒体呼吸、 和免疫相关基因的表达。由于JUUL独特的化学成分，气雾化 参数和喷雾地形，我们预计在JUUL用户中观察到额外的或不同的影响 与上一代电子烟用户相比。我们将在两个特定目标中检验这一假设：SA1将 确定Pod电子烟设备的用户是否在中央呼吸道和 SA2将研究雾化豆荚e液及其成分对巨噬细胞体外功能的影响。 为了实现SA1，来自电子烟使用者(上一代和豆荚队列)的诱导痰样本， 吸烟者和不吸烟者将被分析免疫细胞组成、吞噬功能、 线粒体能量、形态和免疫组织化学染色。诱导痰也将被分析 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。为了实现SA2， 从支气管镜下获得的原代人肺巨噬细胞将被贴附于跨孔和 暴露在空气-液体界面上的气溶胶来自Juul豆荚(烟草和薄荷口味)、尼古丁盐和苯甲酸 使用一种新的体外暴露系统单独使用酸。然后将对细胞进行细胞毒性、炎症性检测 反应，吞噬功能，和生物能量能力，以确定e- 这些终端上的液体。来自这些目标的数据将被集成，以提供高度翻译的、 对豆荚电子烟设备对呼吸免疫功能障碍影响的机械洞察，这将 解决与JUUL等豆荚电子烟的潜在健康影响有关的关键知识差距。

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.