Impact of Peripheral Circadian Misalignment on Resiliency of Intestinal Barrier Function to Alcohol

项目摘要 只有20%-30%的重度酗酒者出现酒精引起的肠道通透性高和酒精性肝 疾病(ALD)。因此，强调发挥关键作用的其他辅助因素是必要的，以更好地 了解发病机制。近年来，昼夜节律的患病率有所上升。 节律紊乱导致人类疾病的发病机制，如结肠高通透性。 然而，外周昼夜节律失调在胃肠道屏障内稳态中的作用 是不完全理解的。我们的重点是与饮酒有关的外围生物钟。 在接近身体的生物休息期进食可能会扰乱周围的昼夜节律。这是错误的时间 进食(WTE)可引起肠道微生物区系及其代谢物的变化，这可能是 肠道易受酒精影响。因此，我们将检验外周昼夜节律紊乱的总体假设 将通过微生物代谢产物降低肠道屏障功能对酒精的弹性。 目标1：通过WTE建立外周昼夜节律紊乱降低结肠的弹性 酒精。我们将扰乱PER2荧光素酶(PER2：：Luc)报告BL/6小鼠的外周昼夜节律 模拟和评估饮酒对肠道通透性和肠屏障功能的影响 目的2：证明细菌代谢物影响外周昼夜节律并调节 结肠屏障功能的弹性降低。我们将证明细菌代谢物可以影响 通过结肠PER2：：Luc调节肠道屏障功能的昼夜节律变化 有机化合物。在这项拟议的研究中，我将扩展关于外周昼夜节律影响的知识 肠道对酒精的弹性失调。此外，我将研究细菌代谢物的影响 来自外周昼夜节律紊乱的PER2：：LUC小鼠。拟议的工作将阐明错误的时间是如何 进食使肠道更容易受到酒精的伤害，这与微生物失调和 细菌产品。成功完成这项研究将极大地提高我们对一个关键问题的理解 酒精引起的屏障功能障碍的辅助因素。最终，结肠外周昼夜节律的建立 ALD的错位和识别与此表型相关的微生物代谢物将导致 制定与微生物和昼夜节律相关的干预措施，并确定治疗目标以缓解 酒精引起的损伤。这项建议将增强受训人员概念化、开发、执行、 并以越来越独立的方式评估有意义的研究问题，将帮助她发展成为 多产、领先的独立研究科学家。研究项目和CAREER的开发将是 由一支杰出的赞助商团队和论文委员会指导，他们非常适合 提供这种指导，例如掌握最先进的方法/技术、指导和 发展协作网络，以确保受训人员在短期和长期取得成功。

Project Abstract Only 20-30% of heavy alcoholics develop alcohol-induced intestinal hyperpermeability and alcoholic liver disease (ALD). Thus, emphasizing other cofactors that play a critical role is necessary to better understand mechanisms behind pathogenesis. Recent years have seen an increased prevalence of circadian rhythm disruption that contributes to human disease pathogenesis such as colonic hyperpermeability. However, the role of peripheral circadian misalignment in barrier homeostasis of t he gastrointestinal tract (GIT) is incompletely understood. Our focus is on the peripheral circadian clock in relation to alcohol consumption. Peripheral circadian disruption can occur by eating close to the body’s biological rest period. This ‘wrong-time eating’ (WTE) can cause changes in the intestinal microbiota and their metabolites, which may mediate intestinal gut leakiness to alcohol. Thus, we will test the overall hypothesis that peripheral circadian disruption will decrease resiliency of the intestinal barrier function to alcohol, mediated through microbial metabolites . Aim 1: Establish that peripheral circadian disruption through WTE decreases resiliency of the colon to alcohol. We will disrupt peripheral circadian rhythms in PERIOD2 luciferase (PER2::Luc) reporter BL/6 mouse model and assess the effect of alcohol consumption on intestinal permeability and intestinal barrier function Aim 2: Demonstrate that bacterial metabolites impact peripheral circadian rhythms and mediate decreased resiliency of colonic barrier function. We will demonstrate that bacterial metabolites can affect circadian rhythms as well as mediate changes in intestinal barrier function through colonic PER2::LUC organoids. In this proposed research, I will expand knowledge regarding the impact of peripheral circadian misalignment on intestinal resiliency to alcohol. Additionally, I will investigate the effect of bacterial metabolites from peripherally circadian disrupted PER2::LUC mice. The proposed work will elucidate how wrong -time eating makes the gut more susceptible to injury by alcohol by mechanisms related to microbial dysbiosis and bacterial products. Successful completion of this research would greatly increase our understanding of a key co-factor in alcohol induced barrier disfunction. Ultimately, the establishment of colonic peripheral circadian misalignment in ALD and identifying the microbial metabolites associated with this phenotype will lead to the development of microbial and circadian related interventions and identify therapeutic targets to mitigate alcohol-induced damage. This proposal will enhance the trainee’s ability to conceptualize, develop, execute, and evaluate meaningful research questions with increasing independence and will help her develop into a productive, leading independent research scientist. The research project and ca reer development will be guided by an outstanding team of sponsors and the dissertation committee who are exceptionally well -suited to provide this guidance such as mastering state-of-the-art methods/techniques, mentorship, and the development of collaborative networks to ensure success of the trainee in the short-term and long-term.