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

项目概要/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， 造成明显的不良健康影响。几种电子液体风味化合物是α，β-不饱和醛， 已知对呼吸道免疫功能有不良影响的化学品类别。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 辛辣风味的电子液体，并具有已知的免疫调节活性。然而，CA的潜在影响 呼吸道免疫应答存在关键的知识缺口，这将是本申请的重点。我们 将使用紧密联系的体外和人体体内研究机制来确定CA对 呼吸道先天免疫功能，特别关注两个组成部分：1）粘膜纤毛组成部分 由衬在气道上的纤毛上皮细胞组成，和2）由常驻和 浸润性白细胞，如巨噬细胞（Macs）。我们的数据表明，含CA的电子液体大大 在不引起明显细胞毒性的剂量下影响纤毛跳动和呼吸免疫细胞功能。这些 这种作用与线粒体呼吸改变有关，并可被巯基还原所抑制 试剂因此，基于现有的知识和我们自己的数据，我们假设含CA的电子液体 通过CA诱导线粒体呼吸和巯基抑制来抑制固有粘膜免疫功能 细胞蛋白质的修饰。我们将在两个具体目标中测试这一假设：SA 1将确定CA- 诱导对上皮纤毛功能和粘膜纤毛清除（MCC）的影响，并确定其机制 调解这些反应。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， 细胞（HBEC）到含CA的电子液体，评估纤毛跳动的变化，并确定 线粒体呼吸和巯基修饰。为了将这些发现转化为人类， 在体内，我们建议有健康的成年志愿者进行控制吸入锝-99m硫 胶体（Tc 99 m-SC）颗粒吸入含CA的电子烟后，然后跟踪 放射性标记的颗粒作为使用γ射线照相术的MCC的量度。SA 2将确定CA诱导的 调节MAC和介导这些反应的机制。为了实现这一目标，我们将 用含有CA的电子液体离体检测人Mac，并检查免疫功能的变化，并确定 巯基修饰和线粒体呼吸在这些反应中的作用。通过诱导获得的Mac 来自经受受控的含CA的电子液体的电子烟暴露的人类受试者的痰（IS）将被 用于将Macs体外获得的机制发现转化为人体体内。得出的数据 这些高度整合的翻译研究将产生重要的机制信息，对流行的电子 香烟增香剂具有对更大组化学增香剂的潜在影响，因此 解决与调味电子烟使用的潜在健康影响相关的临床知识差距。

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.