Airway Epithelial Barrier Dysfunction In Response to Respiratory Syncytial Virus

呼吸道合胞病毒引起的气道上皮屏障功能障碍

基本信息

批准号：
8990806
负责人：
Fariba Rezaee
金额：
$ 17.63万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8990806
关键词：
Actins Acute Address Adherence Adult Advisory Committees Affect Apical Biological Assay Biology Breathing Bronchiolitis Cause of Death Cell Culture Techniques Cell Line Cell physiology Cells Child Childhood Complex Cytoskeleton Data Dedications Development Development Plans Diagnosis Disease Double-Stranded RNA Elderly Elements Endocytosis Endocytosis Pathway Epithelial Epithelial Cells Foundations Functional disorder Goals Human Immune response Immune system Immunology In Vitro Infant Infection Inflammation Mediators Inflammatory Response Injury Knowledge Laboratories Leadership Life Cycle Stages Lower Respiratory Tract Infection Lung Lung Inflammation Lung diseases Mannitol Mediating Mentors Mentorship Modeling Molecular Monitor Morbidity - disease rate Mucous body substance Mus Pathogenesis Pathology Pathway interactions Permeability Physicians Productivity Publications Quality of life Regulation Reporting Research Research Proposals Respiratory Syncytial Virus Infections Respiratory Tract Infections Respiratory syncytial virus Role Scientist Signal Pathway Signal Transduction Signaling Protein Source Structure Surface TLR3 gene Testing Therapeutic Tight Junctions TimeLine Tissues Toll-like receptors Training Translating United States Universities Viral Viral Respiratory Tract Infection Virus Virus Diseases Virus Replication airway epithelium airway inflammation career career development clinically relevant clinically significant collaborative environment design helicase high risk improved in vitro Model in vivo innovation insight mortality mouse model new therapeutic target next generation novel pathogen pediatric department protein complex protein kinase D public health relevance research study respiratory respiratory virus response skills tool virology

项目摘要

DESCRIPTION (provided by applicant): Recent studies indicate that respiratory viruses, including Respiratory Syncytial Virus (RSV), disrupt epithelial barrier structure and function, but the mechanisms involved and consequences for airway inflammation remains poorly understood. The long-term goal of our studies is to better understand how airway epithelial cell barrier structure and function are affected by viral infection and how modifying the involved mechanisms restore barrier integrity and dampen inflammatory responses. Our recent publication revealed that, in cell culture, RSV infection caused disruption of tight junction structure and function. Our objective in this particular application is to investigate the molecula mechanism and signaling pathways involved in RSV-induced barrier disruption. The strength of our approach is that we utilize four highly complementary models including a well-characterized and manipulable human bronchial epithelial cell line, primary human and mouse airway epithelial cells, and cutting-edge mouse models; each serves to examine different key aspects of proposed mechanisms. The central hypothesis is that RSV infection prompts airway epithelial barrier dysfunction via endocytosis of apical junctional complexes (AJC) in a Protein Kinase D (PKD)-dependent manner. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Test the hypothesis that PKD- dependent endocytosis is essential for RSV-induced airway epithelial barrier dysfunction; 2) Test the hypothesis that RSV causes sustained "leaky airway" in a PKD-dependent manner. In Aim 1, we will elucidate endocytic pathways, cytoskeleton remodeling, and PKD activation triggered by RSV. In Aim 2, we will use a well-characterized mouse model of wild type, and cutting-edge conditional deletion of PKD to investigate association of junction dysfunction, lung inflammation, and viral replication. Our proposal is innovative because it is the first comprehensive analysis of role of RSV infection and PKD pathway in the lung. Furthermore, it will provide an innovative assay of outside/in airway permeability by using inhaled mannitol, which has strong translational potential. The proposed research is significant, because it will provide essential knowledge about a clinically relevant virus with poorly understood pathogenesis. The overall K08 application is designed to build a foundation of technical, intellectual and leadership skills required to transition into independence. The career development plan will help me strengthen my knowledge in virology, and barrier immunology, develop expertise in studying cell signaling, and endocytosis, acquire essential skills in utilizing genetically modified mice, and define my niche as an independent physician scientist in the field of respiratory biology. My mentors and advisory committee will monitor adherence to the detailed productivity and developmental progress timeline. My commitments to a research career, strong mentorship, unique dedication of Pediatrics Department in training the next generation of physician-scientists, and outstanding collaborative environment for respiratory virus research at the University of Rochester will provide the necessary tools with which I can build my career.

描述（由适用提供）：最近的研究表明，呼吸道病毒，包括呼吸道合胞病毒（RSV），破坏上皮屏障结构和功能，但涉及的机制和对气道感染的后果仍然鲜为人知。我们研究的长期目标是更好地了解气道上皮细胞屏障结构和功能如何受病毒感染的影响，以及如何修改所涉及的机制如何恢复屏障完整性和该死的感染反应。我们最近的出版物表明，在细胞培养中，RSV感染导致紧密连接结构和功能的破坏。我们在此特定应用中的目标是研究与RSV诱导的屏障破坏有关的分子机制和信号通路。我们方法的优势在于，我们利用了四个高度互补的模型，包括良好的和可操纵的人支气管上皮细胞系，原代人和小鼠气道上皮细胞以及尖端的小鼠模型。每种都用于检查所提出机制的不同关键方面。中心假设是RSV感染通过蛋白激酶D（PKD）依赖性方式的顶端连接络合物（AJC）的内吞作用促使气道上皮屏障功能障碍。在强大的初步数据的指导下，将通过追求两个具体目的来检验该假设：1）检验以下假设：PKD依赖性内吞作用对于RSV诱导的气道上皮屏障功能障碍至关重要； 2）测试以PKD依赖性方式导致RSV导致持续“泄漏气道”的假设。在AIM 1中，我们将阐明由RSV触发的内吞途径，细胞骨架重塑和PKD激活。在AIM 2中，我们将使用良好的野生型小鼠模型和PKD的尖端有条件缺失来研究连接功能障碍，肺部感染和病毒复制的关联。我们的建议具有创新性，因为它是RSV感染和PKD途径在肺中的作用的首次综合分析。此外，它将通过使用内部甘露醇（具有强大的翻译潜力）来提供对气道渗透性外部/渗透率的创新测定。拟议的研究很重要，因为它将提供有关临床相关病毒的基本知识，其发病机理知之甚少。总体K08应用程序旨在建立过渡到独立所需的技术，智力和领导能力的基础。职业发展计划将有助于我加强对病毒学的知识，以及障碍免疫学，发展研究细胞信号的专业知识，内吞作用，获得利用转基因小鼠的基本技能，并将我的利基定义为呼吸生物学领域的独立物理科学家。我的导师和咨询委员会将监视遵守详细的生产力和发展进度时间表的遵守情况。我对研究职业的承诺，强大的指导，儿科系的独特奉献精神在培训下一代的身体科学家以及罗切斯特大学呼吸病毒研究的杰出协作环境中，将提供我可以建立职业生涯的必要工具。