E-cig flavors and their effects on respiratory innate immune responses

项目摘要/摘要 电子烟中的许多调味化学物质都是电子烟特有的一类化学物质，可能 造成明显的不良健康影响。几种电子液体香料化合物是α，β-不饱和醛，a 一类已知对呼吸道免疫功能有不良影响的化学品。在这些调味品中 化合物是肉桂醛(CA)，一种α，β-不饱和醛，通常包含在流行的肉桂或 辛辣口味的电子液体，具有已知的免疫调节活性。然而，CA的潜在影响 呼吸道免疫反应是一个关键的知识缺口，这将是这一应用的重点。我们 将使用紧密联系的体外和人体机制研究来确定CA对 呼吸先天免疫功能，特别关注两个组成部分：1)粘液纤毛成分 由排列在呼吸道内的纤毛上皮细胞组成，2)细胞成分，由滞留和 渗透的白细胞，如巨噬细胞(Mac)。我们的数据表明，含CA的电子液体极大地 影响纤毛跳动和呼吸免疫细胞功能的剂量不会造成明显的细胞毒性。这些 这种作用与线粒体呼吸的改变有关，并可被硫醇还原所抑制 试剂。因此，根据现有知识和我们自己的数据，我们假设含有CA的电子液体 CA抑制线粒体呼吸和硫醇抑制粘膜天然免疫功能 细胞蛋白质的修饰。我们将在两个具体目标中检验这一假设：SA1将决定CA- 对上皮纤毛功能和粘液纤毛清除的诱导作用及其机制探讨 调停这些反应。为了实现这一目标，我们将暴露分化良好的人支气管上皮 细胞(HBECs)到含CA的电子液体，评估纤毛搏动的变化，并确定 线粒体呼吸和这些反应中的硫醇修饰。将这些发现转化为人类 体内，我们建议让健康的成年志愿者接受~(99m)Tcs的控制吸入 吸入含CA电子烟后的胶体(Tc99m-Sc)颗粒，随后追踪 使用伽马闪烁照相术将放射性标记的粒子作为MCC的测量。SA2将确定CA诱导的 MACs的调节和调节这些反应的机制。为了实现这一目标，我们将刺激 人的Mac与含有CA的电子液体体外检测免疫功能的变化，并确定 硫醇修饰和线粒体呼吸在这些反应中的作用。通过诱导获得MACs 接受受控蒸发暴露于含CA电子液体的受试者的痰(IS)将被 用于将在MACS体外获得的机制发现转化为体内的人类。数据源自 这些高度集成的翻译研究将在流行的电子邮件中产生重要的机械信息- 卷烟调味剂可能涉及更多的化学调味剂，因此 解决与使用调味电子烟的潜在健康影响有关的临床知识差距。

Project Summary/Abstract Many of the flavoring chemicals in e-cigarettes present a class of chemicals unique to e-cigarettes, potentially causing distinct adverse health effects. Several e-liquid flavoring compounds are α,β-unsaturated aldehydes, a class of chemicals with known adverse effects on respiratory immune function. Among those flavoring compounds is cinnamaldehyde (CA), an α,β-unsaturated aldehyde often contained in popular cinnamon or spicy flavored e-liquids and with known immune modulating activities. However, potential effects of CA on respiratory immune responses present a critical knowledge gap, which will be the focus of this application. We will use tightly linked mechanistic in vitro and human in vivo studies to determine adverse effects of CA on respiratory innate immune functions, with specific focus on two components: 1) the mucociliary component consisting of ciliated epithelial cells lining the airways and 2) the cellular component consisting of resident and infiltrating leukocytes, such as macrophages (Macs). Our data demonstrate that CA-containing e-liquids greatly affect ciliary beating and respiratory immune cell function at doses that do not cause overt cytotoxicity. These effects were associated with modified mitochondrial respiration and could be inhibited by thiol reducing reagents. Thus, based on existing knowledge and our own data we hypothesize that CA-containing e-liquids suppress innate mucosal immune function by CA-induced inhibition of mitochondrial respiration and thiol modification of cellular proteins. We will test this hypothesis in two specific aims: SA1 will determine CA- induced effects on epithelial ciliary function and mucociliary clearance (MCC) and identify the mechanisms mediating these responses. To achieve this aim we will expose well-differentiated human bronchial epithelial cells (HBECs) to CA-containing e-liquids, assess changes in ciliary beating, and determine the role of mitochondrial respiration and thiol modification in these responses. To translate these findings into humans in vivo, we propose to have healthy adult volunteers undergo controlled inhalation of Technetium-99m sulfur colloid (Tc99m-SC) particles after inhalation of CA-containing e-cigarettes, followed by tracking the egress of the radiolabeled particles as a measure of MCC using gamma scintigraphy. SA2 will determine CA-induced modulation of Macs and the mechanisms mediating these responses. To achieve this aim we will stimulate human Macs with CA-containing e-liquids ex vivo and examine changes in immune function, and determine the role of thiol modification and mitochondrial respiration in these responses. Macs obtained through induced sputum (IS) from human subjects undergoing controlled vaping exposure to CA-containing e-liquids will be used to translate the mechanistic findings obtained in Macs ex vivo into humans in vivo. Data derived from these highly integrated translational studies will yield important mechanistic information on a popular e- cigarette flavoring agent with potential implications for a larger group of chemical flavoring agents, thus addressing a clinical knowledge gap related to the potential health effects of flavored e-cigarette use.