Ethanol-mediated cilia motility dysfunction

描述（由申请人提供）：已知大量饮酒会损害肺部保护自身免受肺炎和支气管炎等感染的能力。肺部抵御此类感染的第一道防线是利用纤毛粘液装置捕获吸入颗粒并将其排出肺部。在一个健康的个体中，这个器官产生粘液，它捕获吸入的颗粒，然后被纤毛（肺内膜的指状突起）推进出肺。纤毛受损会导致肺炎和肺功能丧失。我们已经确定，酒精通过改变一种叫做激酶的关键蛋白质，对这种脆弱的粘液纤毛器官造成了严重伤害。其中两种激酶，PKA和PKG，是调控包括肺纤毛在内的细胞功能的分子。我们在酒精喂养的小鼠中进行的实验表明，酒精严重损害了这些激酶的功能，导致纤毛粘膜器官正常的“战斗或逃跑”反应丧失，而这种反应需要产生一氧化氮。虽然我们已经确定酒精会导致这个问题，但我们不知道如果去除酒精，这个问题会持续多久。我们也不知道酒精是如何改变纤毛中这些重要的激酶依赖功能的。这导致我们假设：慢性酒精暴露通过改变气道上皮纤毛中的特定调节蛋白导致纤毛粘膜功能的时间依赖性和可逆性损伤。在这个提案中，我们提出了旨在回答两个问题的实验：1。酒精引起的纤毛粘膜功能损害是可以预防的还是可逆的？和2。哪些纤毛蛋白受到酒精的影响？我们建议通过以下四个具体目标提出的实验来回答这些问题：1。描述慢性酒精损伤纤毛粘膜功能的时间过程和可逆性2. 明确氧化应激在酒精介导的一氧化氮依赖性纤毛运动调节中的信号通路和作用，3。确定存在于被酒精暴露改变的纤毛上的激酶磷酸化靶标；和4。使用分离的纤毛模型确定酒精介导的纤毛损伤的分子决定因素。虽然我们已经了解了酒精损伤对纤毛粘膜功能的影响和机制，但我们提出的研究将扩展我们的知识，了解如何预防或治疗这种情况，以及酒精如何改变纤毛功能和肺部健康的关键分子。

DESCRIPTION (provided by applicant): Heavy alcohol intake is known to damage the ability of the lung to protect itself from infections such as pneumonia and bronchitis. The lung's first line of defense against such infection is to trap inhaled particles and propel them out of the lung using the mucociliary apparatus. In a healthy individual, this apparatus produces mucus, which traps inhaled particles, and is then propelled out of the lung by cilia, the fingerlike projections of the lung lining. Damage to cilia results in pneumonia and loss of lung function. We have established that alcohol profoundly injures this delicate mucociliary apparatus by altering critical proteins called kinases. Two of these kinases, PKA and PKG, are well-described molecules that regulate cell functions including lung cilia. Our experiments in alcohol-fed mice have demonstrated that alcohol profoundly impairs the function of these kinases, resulting in a loss of the normal "fight or flight" responsiveness of the mucociliary apparatus in a manner that requires the production of nitric oxide. While we have established that alcohol causes this problem, we do not know how long it persists if alcohol is removed. We also do not know exactly how alcohol alters these important kinase-dependent functions in cilia. This leads us to hypothesize that: Chronic alcohol exposure causes time-dependent and reversible impairment of mucociliary function by altering specific regulatory proteins in airway epithelial cilia. In this proposal, we propose experiments designed to answer two questions: 1. Is alcohol-driven impairment of mucociliary function preventable or reversible? and 2. Which cilia proteins are affected by alcohol? We propose to answer these questions through experiments proposed in four specific aims: 1. Characterize the time course and reversibility of chronic alcohol impairment of mucociliary function; 2. Define the signaling pathways and role that oxidant stress plays in alcohol-mediated nitric oxide-dependent regulation of ciliary motility, 3. Determine the kinase phosphorylation targets present on cilia that are altered by alcohol exposure; and 4. Identify the molecular determinants of alcohol-mediated cilia injury using isolated cilia models. Although we have learned much about the impact and mechanisms of alcohol injury to mucociliary function, the studies we propose will extend our knowledge into how this may be prevented or treated and exactly how alcohol alters molecules critical for cilia function and lung health.