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，人们都知道酒精滥用会损害肺部宿主的防御能力，增加重度饮酒者患支气管炎、肺炎和急性呼吸窘迫综合征的风险。我们的实验室专注于酒精对肺传导气道中的粘膜纤毛清除的影响，这提供了肺对抗吸入的感染因子、颗粒和碎片的第一道防线。我们已经建立了两个关键的观察，推动这一建议：1）短暂的适度饮酒刺激气道纤毛运动; 2）持续大量饮酒损害粘膜纤毛清除，增加气道损伤和肺炎的风险。我们将这种损害称为“酒精诱导的睫状体功能障碍”（AICD）。我们的机制研究表明，短暂的酒精刺激和持续的AICD涉及纤毛细胞机制依赖于一氧化氮（NO），纤毛相关环化酶和环核苷酸依赖性激酶（PKG和PKA）的调节。最近，我们发表了这些酒精触发的纤毛调节途径明显定位于每个气道纤毛的基体中，我们称之为酒精响应的“纤毛代谢子”。该途径对非常低浓度的酒精（1-10 mM，小于法律的中毒极限的一半）非常敏感，并且在分离的气道纤毛细胞器中完全起作用。这使我们假设：酒精导致可逆的气道纤毛功能障碍，由于修改一氧化氮信号和改变的关键cila蛋白的调节，包括动力蛋白马达和调节动力蛋白的机制。在这个提议中，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NO的产生？我们在目标#1中通过研究酒精通过增强热休克蛋白90（HSP 90）的eNOS伴侣功能在激活一氧化氮合酶（eNOS）中所起的作用来探索这个问题。2)为什么持续的酒精暴露会消耗睫状体NO水平？我们在目标#2中通过定义eNOS如何由于L-精氨酸消耗和活性氧物质（ROS）的产生而通过持续的酒精暴露而解偶联来解决这个问题。我们认为纤毛ROS的增加导致eNOS辅因子四氢生物蝶呤（BH 4）的耗竭，从而导致纤毛脱敏。3)纤毛调节酶如何相互作用以改变运动性？在目标#3中，我们将确定持续的酒精暴露如何激活蛋白磷酸酶1（PP 1），从而导致HSP 90和其他关键纤毛激活蛋白（如PKA）的去磷酸化。4)酒精暴露会改变哪些下游纤毛运动分子？使用酒精敏感的能动生物体衣原体的模型遗传系统，目标#4将专注于持续的酒精如何改变纤毛外动力蛋白臂，这是使纤毛跳动的马达蛋白。我们提出的研究将大大扩展我们对如何预防和治疗AICD的知识，并将扩大我们对酒精如何改变对气道纤毛功能至关重要的纤毛分子的了解。1 公共卫生关系：纤毛是支气管的指状突起，可以产生波浪，清除肺部的粘液和吸入的颗粒，通常在压力事件中被激活，以更快地清除粘液。感冒、食物窒息或呼吸脏空气会导致纤毛产生一氧化氮，使它们跳动得更快以清除肺部。大量饮酒后，肺纤毛对压力反应迟钝，因为它们不再产生一氧化氮，导致粘液充血和感染，如支气管炎和肺炎。我们的研究重点是酒精如何关闭肺纤毛产生一氧化氮，重点是发现逆转和/或预防酒精相关纤毛损伤的方法，以避免肺部感染。1