Ethanol-mediated cilia motility dysfunction

乙醇介导的纤毛运动功能障碍

• 批准号: 8372198
• 负责人: Joseph H Sisson
• 金额: $ 48.27万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8372198
• 关键词: Address; Adult Respiratory Distress Syndrome; Affect; Air; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcoholic beverage heavy drinker; Alcohols; Antioxidants; Arginine; Breathing; Bronchitis; Cells; Chlamydomonas; Choking; Cilia; Cyclic AMP-Dependent Protein Kinases; Cyclic Nucleotides; Dynein ATPase; Electron Spin Resonance Spectroscopy; Enzymes; Ethanol; Event; Fingers; Food; Functional disorder; Generations; Genetic; Genetic Models; Heat-Shock Proteins 90; Host Defense; Human; Impairment; Individual; Infection; Infectious Agent; Intoxication; Knowledge; Laboratories; Lead; Legal; Life; Link; Lung; Measurement; Mediating; Modeling; Molecular Chaperones; Motion; Motor; Mucociliary Clearance; Mucous body substance; Mus; Nitric Oxide; Nitric Oxide Synthase; Organelles; Organism; Orthologous Gene; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Pneumonia; Prevention strategy; Production; Protein Dephosphorylation; Protein phosphatase; Proteins; Publishing; Reactive Oxygen Species; Regulation; Research; Risk; Role; Signal Transduction; Slice; Stress; Supplementation; System; Testing; Tube; alcohol effect; alcohol exposure; alcohol use disorder; arm; cell motility; cilium motility; cofactor; desensitization; design; feeding; gene discovery; human NOS3 protein; injured airway; innovation; insight; kinetosome; lung basal segment; motor control; novel; oxidant stress; particle; prevent; protein activation; research study; tetrahydrobiopterin; treatment strategy

DESCRIPTION (provided by applicant): Alcohol abuse has long been known to impair lung host defenses increasing the risk in heavy drinkers for bronchitis, pneumonia and acute respiratory distress syndrome. Our laboratory has focused on alcohol's impact on mucociliary clearance in the conducting airways of the lung, which provides the first line of defense of the lung against inhaled infectious agents, particles and debris. We have established two key observations that drive this proposal: 1) brief modest alcohol intake stimulates airway ciliary motility; and 2) sustained heavy alcohol intake impairs mucociliary clearance increasing the risk for airway injury and pneumonia. We have termed this impairment "Alcohol-Induced Ciliary Dysfunction" (AICD). Our mechanistic studies have demonstrated that both the brief alcohol stimulation and sustained AICD involve ciliated cell mechanisms dependent on the regulation of nitric oxide (NO), cilia-associated cyclases and cyclic nucleotide-dependent kinases (PKG & PKA). Recently, we published that these alcohol-triggered ciliary regulation pathways are distinctly localized to the basal body of each airway cilium in what we have called the alcohol-responsive "ciliary metabolon". This pathway is exquisitely sensitive to very low concentrations of alcohol (1-10 mM, which is less than half of the legal intoxication limit) and is fully functionl in isolated airway cilia organelles. This leads us to hypothesize that: Alcohol causes reversible airway ciliary dysfunction due to modified nitric oxide signaling and altered regulation of key cila proteins, including the dynein motors and the mechanisms that regulate dyneins. In this proposal, we outline experiments designed to answer four new questions: 1) How does brief alcohol exposure stimulate NO production in airway cilia? We explore this question in Aim #1 by examining the role alcohol plays in activating nitric oxide synthase (eNOS) by enhancing the chaperone function of eNOS by heat shock protein 90 (HSP90). 2) Why does sustained alcohol exposure deplete ciliary NO levels? We address this question in Aim #2 by defining how eNOS becomes uncoupled by sustained alcohol exposure due to L- arginine depletion and the generation of reactive oxygen species (ROS). We think increased ciliary ROS leads to depletion of the eNOS cofactor tetrahydrobiopterin (BH4) resulting in cilia desensitization. 3) How do cilia regulatory enzymes interact to modify motility? In Aim #3 we will determine how sustained alcohol exposure activates protein phosphatase 1 (PP1), which leads to dephosphorylation of HSP90 and other key cilia activation proteins such as PKA. 4) What downstream cilia motor molecules are modified by alcohol exposure? Using the model genetic system of the alcohol-sensitive motile organism, Chlamydomonas, Aim #4 will focus on how sustained alcohol modifies ciliary outer dynein arms, which are the motor proteins that make cilia beat. The studies we propose will greatly extend our knowledge of how to prevent and treat AICD and will expand our insight into how alcohol alters cilia molecules critical for airway cilia function. 1 PUBLIC HEALTH RELEVANCE: Cilia, the wave-producing finger-like projections of the bronchial tubes that clear mucus and inhaled particles from the lungs, are normally activated by stress events to clear mucus faster. Catching a cold, choking on food or breathing in dirty air cause cilia to produce nitric oxide, which makes them beat faster to clear out the lungs. After heavy alcohol drinking, lung cilia become unresponsive to stress, because they can no longer make nitric oxide, which leads to mucus congestion and infections such as bronchitis & pneumonia. Our research is focused on how alcohol shuts off nitric oxide production by lung cilia with an emphasis on discovering ways to reverse and/or prevent alcohol-related cilia damage to avoid lung infections. 1

描述（由申请人提供）：长期以来，众所周知，酗酒会损害肺部防御，从而增加了支气管炎，肺炎和急性呼吸窘迫综合征的重量饮酒者的风险。我们的实验室专注于酒精对肺部导电道的粘膜纤毛清除率的影响，该气道提供了对肺部对吸入的感染剂，颗粒和碎屑的第一道防御。我们已经建立了两个推动这一建议的关键观察：1）简短的适度酒精摄入刺激气道纤毛运动； 2）持续的大量酒精摄入会损害粘膜纤毛间隙，从而增加了气道受伤和肺炎的风险。我们称这种障碍为“酒精引起的睫状功能障碍”（AICD）。我们的机械研究表明，短暂的酒精刺激和持续的AICD涉及纤毛的细胞机制，取决于一氧化氮（NO），纤毛相关的环化酶和环状核苷酸依赖性激酶（PKG＆PKA）的调节。最近，我们发表了这些被酒精触发的睫状调节途径明显地定位于每个气道纤毛的基体中，我们称为酒精反应性的“纤毛变质”。该途径对非常低浓度的酒精（1-10毫米，小于法律中毒限制的一半）非常敏感，并且在孤立的气道纤毛细胞器中充分发挥作用。这使我们假设：酒精会导致可逆的气道睫状功能障碍，这是由于一氧化氮信号传导和关键CILA蛋白的调节而改变的，包括动力蛋白电动机以及调节动力蛋白的机制。在此提案中，我们概述了旨在回答四个新问题的实验：1）短暂的酒精暴露如何刺激气道纤毛中没有生产？我们通过检查酒精在激活一氧化氮合酶（ENOS）中的作用来探讨这个问题，通过通过热休克蛋白90（HSP90）增强eNOS的伴侣功能。 2）为什么持续的酒精暴露会耗尽睫状体无水平？我们通过定义eNOS如何因l-精氨酸耗竭而导致的持续酒精暴露并产生活性氧（ROS）来解决AIM＃​​2中的这个问题。我们认为，增加的睫状ROS会导致eNOS辅因子四氢无生蛋白（BH4）的耗竭，从而导致纤毛脱敏。 3）纤毛调节酶如何相互作用以修饰运动性？在AIM＃3中，我们将确定持续酒精暴露如何激活蛋白质磷酸酶1（PP1），这导致HSP90和其他关键的CILIA激活蛋白（如PKA）的去磷酸化。 4）哪些下游纤毛运动分子是通过酒精暴露来改变的？使用对酒精敏感的运动生物的模型遗传系统，AIM＃4将集中于持续酒精如何修饰纤毛外动臂，这些纤毛外动臂是使纤毛跳动的运动蛋白。我们提出的研究将大大扩展我们如何预防和治疗AICD的知识，并将扩展我们对酒精如何改变纤毛分子对气道纤毛功能至关重要的见解。 1 公共卫生相关性：纤毛是支气管中产生的手指样刺激性的，这些指标通常会通过压力事件来激活肺部的粘液和吸入的颗粒，以更快地清除粘液。抓住食物或肮脏的空气呼吸的感冒，使纤毛产生一氧化氮，这使得它们更快地击败以清除肺部。大量饮酒后，肺纤毛对压力无反应，因为它们不能再产生一氧化氮，这会导致粘液充血和感染，例如支气管炎和肺炎。我们的研究集中在肺纤毛中酒精如何关闭一氧化氮的产生，重点是发现逆转和/或防止酒精相关的纤毛损害以避免肺部感染的方法。 1