Aerosolized Nicotine Modulation of Host Inflammation and Microbiota Dysbiosis

雾化尼古丁产品的使用，主要是电子尼古丁输送系统（ENDS），正在增长 尤其是在以前不一定使用烟草产品的年轻人中。 我们对这对他们未来健康状况的影响知之甚少。对我们理解的挑战 ENDS对健康的独特关注包括其高度可变和不一致的配方。尽管如此 在可变性方面，ENDS共同具有生物活性组分尼古丁的递送，尼古丁存在于 纯度和浓度较高。该提案将重点研究尼古丁的生物学效应， 是通过雾化（aeroNic）和吸入递送所特有的。我们小组研究了尼古丁对 外周免疫和中枢过程导致成瘾，因为最初克隆的烟碱 乙酰胆碱受体（nAChR）。由于尼古丁的作用高度依赖于其传递途径，我们 首先开发了一种可靠的小鼠航空给药方法，可产生定量摄取， 动力学与人类相似。实验重点将应用这种AeroNic输送系统来定义 其对小鼠肺部和胃肠道（GI）炎症停滞的影响，并确定这如何改变 急性肺损伤（ALI）或过敏性嗜酸性粒细胞性肺炎（AFL）引起的炎症反应 炎症（AEI;哮喘模型）。遗传工具的应用将极大地促进这一分析 通过尼古丁在外周细胞中的主要靶点nAChRalpha 7（α7）来操纵信号。这 包括尼古丁如何耦合到特定的钙信号网络来调节这些促炎反应。 在初步研究中，通过肺中α7钙偶联机制的呼吸作用减少了炎症反应。 反应和改变上皮细胞信号网络，如那些控制粘蛋白的生产。最近 我们已经发现aeroNic对微生物群生态失调的同时和强大的影响。提出的实验 基于这些初步和已发表的研究结果来测试该项目的假设：雾化尼古丁的作用是 通过调节α7钙信号网络，降低肺对ALI和AEI的反应性 免疫-上皮-微生物群相互作用的正常调节。这将在 三个具体目标。目标1将测量aeroNic如何改变小鼠微生物群，以及微生态失调是否 永久的目的2将定义转录信号网络和蛋白质组学机制，通过这些机制，aeroNic 通过α7改变正常上皮细胞功能。目标3将定义aeroNic如何修改肺/GI轴 通过已知调节ALI和AEI的粘膜免疫细胞的修饰来抑制停滞。结束时 通过这些实验，我们将清楚地了解aeroNic对肺和胃肠道的独特生物学影响。 以及这些变化如何改变长期健康结果。

The use of aerosolized nicotine products, mostly electronic nicotine delivery systems (ENDS), are growing in popularity and especially among younger individuals who have not necessarily used tobacco products previously. We know little about the impact this has on their future health landscape. Challenges to our understanding of the unique concerns ENDS present to health include their highly variable and inconsistent formulations. Despite this variability, ENDS share in common the delivery of the biologically active component nicotine, which is present in relatively high purity and concentration. This proposal will focus on examining the biologic effects of nicotine that are unique to delivery by aerosolization (aeroNic) and inhalation. Our group has studied the impact of nicotine on both peripheral immune and central processes leading to addiction since the original cloning of the nicotinic acetylcholine receptors (nAChR). Because the effects of nicotine are highly dependent upon its route of delivery, we have first developed a reliable method of aeroNic administration to the mouse that produces quantitative uptake and kinetics comparable to those in humans. The experimental focus will apply this AeroNic delivery system to define its impact on mouse inflammatory stasis in the lung and gastrointestinal (GI) tract, and determine how this modifies the inflammatory response to challenge of the lung by either acute lung injury (ALI) or to allergic eosinophilic inflammation (AEI; a model of asthma). This analysis will be greatly facilitated through application of genetic tools that manipulate signaling through nicotine's principal target in peripheral cells, the nAChRalpha7 (α7). This includes how nicotine couples to specific calcium signaling networks to modulate these pro-inflammatory responses. In preliminary studies, aeroNic actions through α7 calcium-coupled mechanisms in the lung reduce inflammatory responsiveness and alter epithelial cell signaling networks such as those controlling mucin production. Most recently we have discovered a concurrent and robust impact by aeroNic on microbiota dysbiosis. The experiments proposed build upon these preliminary and published findings to test the project hypothesis: Aerosolized nicotine acts to depress lung responsiveness to ALI and to AEI through modifying α7 calcium signaling networks controlling normal modulation of immune - epithelial - microbiota interactions. This will be tested in experiments outlined in three interactive Specific aims. Aim 1 will measure how aeroNic alters the mouse microbiota and if dysbiosis is permanent. Aim 2 will define transcriptional signaling networks and proteomic mechanisms through which aeroNic acts through α7 to modify normal epithelial cell function. Aim 3 will define how aeroNic modifies the lung/GI axis stasis through modifications of mucosal immune cells known to regulate both ALI and AEI. At the conclusion of these experiments we will have a clear understanding of the unique biological impact of aeroNic on the lung and GI and how these changes may modify long-term health outcomes.