Autophagy regulates airway epithelial cell mucin secretion

自噬调节气道上皮细胞粘蛋白分泌

• 批准号: 9087595
• 负责人: John David Dickinson
• 金额: $ 15.98万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9087595
• 关键词: Air; Asthma; Autophagocytosis; Autophagosome; Biology; Calcium; Cells; Cellular Stress; Chronic; Chronic Obstructive Airway Disease; Clinical; Collaborations; Cystic Fibrosis; Data; Development; Development Plans; Disease; Ensure; Epithelial; Epithelial Cells; Eukaryota; Future; Generations; Goals; Health; Homeostasis; Human; In Vitro; Inflammation; Inflammatory; Interleukin-13; Journals; Knowledge; Liquid substance; MUC5AC gene; MUC5B gene; Measures; Mediating; Membrane; Mentors; Mentorship; Metaplasia; Mitochondria; Modeling; Morbidity - disease rate; Mucins; Mucous body substance; Mus; NADPH Oxidase; Nebraska; Nutrient; Organelles; Pathway interactions; Phenotype; Physicians; Play; Production; Protein Secretion; Proteins; Publishing; Reactive Oxygen Species; Recycling; Regulation; Research; Research Personnel; Resources; Role; Scientist; Signal Transduction; Source; Stress; Structure; Testing; Therapeutic; Thick; Time; Tracheobronchial; Universities; Work; airway epithelium; airway inflammation; airway mucous cell metaplasia; airway obstruction; biological adaptation to stress; career; career development; cytokine; experience; extracellular; genetic regulatory protein; human disease; in vivo; in vivo Model; inflammatory lung disease; meetings; mortality; mouse model; novel; novel therapeutic intervention; oxidant stress; programs; protein transport; public health relevance; research study; response

 DESCRIPTION (provided by applicant): Significance of airway disease: Airway disease including asthma, chronic obstructive pulmonary disease, and cystic fibrosis are a significant cause of morbidity and mortality. A key feature of chronic inflammatory airway disease is hyper secretion of mucin proteins and airway obstruction by thick tenacious mucus. How the airway epithelial cell adapts to this stress of increased production and secretion of airway secretory mucins is not well understood. Autophagy as a novel pathway to regulate secretion: Autophagy, while highly conserved amongst eukaryotes, was previously thought to primarily regulate nutrient recycling in times of stress. It has now been established that autophagy is essential for multiple responses including protein secretion. Furthermore, autophagy appears to be particularly required during periods of high protein production and secretion during cellular stress. Our data just published in the Journal, Autophagy6, demonstrates that in the airway epithelia, autophagy is required for secretion of MUC5AC during Th2 inflammation. However, the mechanism of autophagy regulated mucin secretion in the airway is not known. Research Plan: In this application we present our preliminary data using in vitro human epithelial cells and in vivo mouse models of Th2 inflammation, demonstrating that autophagy is required for both MUC5Ac secretion and reactive oxygen species (ROS) generation. We hypothesize that these shared phenotypes are in fact connected and that autophagy-dependent ROS may be a novel mechanism that regulates secretion. To test this hypothesis, first, we will examine the role of calcium in autophagy-mediated basal and ATP-activated secretion of the secretory mucins, MUC5AC and MUC5B in the absence and presence of IL-13 using primary human airway epithelial cells (hTEC). Second, we will determine whether autophagy-dependent ROS regulates MUC5AC and or MUC5B independent of mucin production using hTEC. Third, we will examine the relationship among autophagy, ROS, and mucin secretion in an in vivo Th2 inflammation model using the autophagy deficient mice, Atg16l1HM/HM. This research application will advance our knowledge of airway biology using relevant models of human disease with the goal of identifying new therapeutic strategies. This work will advance through a structured program of career development with a goal of becoming an independent investigator. Career Development: As a physician-scientist, it is critically important to have strong mentorship from experienced investigators. My primary mentor, Dr. Joseph Sisson is fully committed to support my career goal of becoming an independent investigator in airway diseases. He has provided resources to establish my lab, protect my research time with limited clinical duties, so that I have begun work on experiments outlined above within the context of a career development plan. This plan includes oversight by experienced investigators on a formal mentorship committee to ensure that important career milestones are being met, formal course work, and development of new collaborations at the University of Nebraska and around the world.

 描述(由申请人提供)：呼吸道疾病的意义：包括哮喘、慢性阻塞性肺疾病和囊性纤维化在内的呼吸道疾病是发病率和死亡率的重要原因。慢性炎症性呼吸道疾病的一个主要特征是粘蛋白的过度分泌和粘稠粘液造成的呼吸道阻塞。呼吸道上皮细胞如何适应这种增加的呼吸道分泌性粘蛋白的产生和分泌的压力还不是很清楚。自噬作为一种调节分泌的新途径：自噬虽然在真核生物中高度保守，但以前被认为主要调控应激时期的营养循环。现在已经证实，自噬对于包括蛋白质分泌在内的多种反应是必不可少的。此外，在细胞应激期间蛋白质产生和分泌高峰期，自噬似乎尤其必要。我们刚刚发表在《自噬》杂志上的数据表明，在Th2炎症期间，在呼吸道上皮细胞中，MUC5AC的分泌需要自噬。然而，自噬调节呼吸道粘蛋白分泌的机制尚不清楚。研究计划：在这项应用中，我们使用体外培养的人上皮细胞和 在活体小鼠的Th2炎症模型中，证明自噬对于MUC5AC的分泌和活性氧物种(ROS)的产生都是必需的。我们假设这些共同的表型实际上是相互联系的，依赖自噬的ROS可能是一种调节分泌的新机制。为了验证这一假设，首先，我们将利用原代人呼吸道上皮细胞(HTEC)，在没有和存在IL-13的情况下，研究钙在自噬介导的基础和ATP激活的分泌性粘蛋白、MUC5AC和MUC5B分泌中的作用。其次，我们将确定自噬依赖的ROS是否独立于使用hTEC调节粘蛋白产生而调节MUC5AC和/或MUC5B。第三，我们将使用自噬缺陷小鼠Atg16l1HM/HM在体内Th2炎症模型中检查自噬、ROS和粘蛋白分泌之间的关系。这项研究应用将利用人类疾病的相关模型来促进我们对呼吸道生物学的了解，目的是确定新的治疗策略。这项工作将通过一个结构化的职业发展计划来推进，目标是成为一名独立的调查员。职业发展：作为一名内科科学家，拥有经验丰富的研究人员的强大指导至关重要。我的主要导师约瑟夫·西森博士全力支持我成为一名呼吸道疾病独立研究员的职业目标。他提供了资源来建立我的实验室，用有限的临床职责保护我的研究时间，所以我已经开始在职业发展计划的背景下进行上面概述的实验工作。该计划包括由经验丰富的调查人员在正式的指导委员会上进行监督，以确保实现重要的职业里程碑，正式的课程工作，以及在内布拉斯加大学和世界各地发展新的合作。