Mechanisms of Airway Epithelial Barrier Dysfunction by Respiratory Syncytial Virus and Environmental Stimuli

呼吸道合胞病毒和环境刺激导致气道上皮屏障功能障碍的机制

基本信息

批准号：
10657436
负责人：
Fariba Rezaee
金额：
$ 43.99万
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：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10657436
关键词：
3-Dimensional Actin-Binding Protein Actins Acute Adult Adverse effects Affect Age Air Pollution Airway Disease Antioxidants Apical Artificial nanoparticles Attenuated Cell Separation Child Childhood Complex Cyclic AMP Cytoskeleton Data Deposition Distal Down-Regulation EMS1 gene Environmental Pollutants Environmental Pollution Epidemiology Epithelial Cells Epithelium Event Exposure to Filament Free Radicals Functional disorder Generations Hospitalization Host Defense Household Human Human Cell Line In Vitro Infant Inflammation Injury Knockout Mice Link Lower Respiratory Tract Infection Lung Measurement Mediating Methods Microfilaments Mission Modeling Molecular Monomeric GTP-Binding Proteins Morbidity - disease rate Mus Nanotechnology Organoids Outcome Oxidative Stress Oxidative Stress Induction Particle Size Particulate Matter Pathology Pathway interactions Personal Satisfaction Polymers Production Reporting Research Respiration Disorders Respiratory Syncytial Virus Infections Respiratory syncytial virus Risk Rodent Role Severities Signaling Molecule Source Standardization Stimulus Structure Testing Therapeutic Intervention United States National Institutes of Health Vascular Endothelium Vascular Permeabilities Virus Virus Diseases Virus Replication airway epithelium airway hyperresponsiveness bronchial epithelium clinically relevant commercial application consumer product depolymerization design epithelial injury gain of function high risk in vivo in vivo Model injured airway innovation insight intestinal epithelium loss of function mortality mouse model nanomaterials nanoparticle new therapeutic target novel particle polymerization titanium dioxide young adult

项目摘要

ABSTRACT Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory tract infections (ALRI) in children and high-risk adults worldwide. Our data demonstrate that RSV infection results in a ‘leaky airway’ by disrupting epithelial apical junctional complexes (AJC), which regulate the airway epithelial barrier. We show that RSV- mediated disruption of AJC is accompanied by disassembly of the perijunctional actin cytoskeleton, and downregulation of cortactin - a key actin-binding protein. Cortactin deficiency has been previously implicated in destabilizing the vascular endothelial and intestinal epithelial barrier. However the causal link between RSV- induced leaky barrier, actin cytoskeletal rearrangements and cortactin deficiency have not been established. In addition, epidemiological reports suggest a strong association between exposure to ambient particulate matter (PM) and increased risk of ALRI. Nanoparticles (NPs) are extremely small PM, with the greater ability to become deposited in distal airways and evade host defenses compared to smaller particles. Our preliminary data demonstrate that pre-exposure of bronchial epithelial cells to NP not only enhances RSV-induced AJC disassembly and actin cytoskeleton disruption, but augments viral infection. Based on our novel observations, we formulated the central hypotheses that a) RSV induces disruption of the airway epithelial barrier by triggering depolymerization of the perijunctional actin cytoskeleton, and by downregulating cortactin; and b) that disruption of the epithelial barrier by nanoparticles worsens RSV-induced airway epithelium injury. We will test our hypotheses through the following Specific Aims: Aim 1: To determine the role of cortactin-dependent actin filament dynamics in RSV-induced airway epithelial barrier dysfunction. Using human bronchial epithelial cells isolated from pediatric donors, and a mouse model of cortactin null mice, we will investigate (i) RSV effect on actin cytoskeletal dynamics, (ii) the functional role of cortactin on actin dynamics and AJC structure, and (iii) the functional roles of Rap-1. We will also use a 3-D human lung organoids model detailing the effects of AJC disruption upon RSV infection, which offers an innovative platform to study complex host-environmental interactions. Aim 2: To determine if nanoparticles enhance RSV-induced disruption of the airway epithelial barrier. Using in vitro and in vivo models, we will (i) characterize the effects of particle size on barrier integrity, (ii) study role of oxidative stress on AJC function, and (iii) define the effects of exposure to NP on AJC dysfunction. The proposed research is significant and relevant to the NIH’s mission as we aim to explore the clinically relevant consequences of RSV infection on airway barrier integrity, and how exposure to environmental pollutants worsens RSV infection. Our approach is innovative because it will provide new mechanistic insight in to the roles of RSV in AJC disassembly, as well as novel insight in to the effects of NPs in enhancing RSV- induced AJC disruption. The identified pathways will provide new targets for therapeutic intervention and the potential for positively impacting the management of RSV disease.

抽象的呼吸综合病毒（RSV）是儿童急性下呼吸道感染（ALRI）的主要原因和全球高风险的成年人。我们的数据表明，RSV感染通过中断导致“漏气道” 上皮顶连接络合物（AJC），该复合物调节气道上皮屏障。我们表明RSV- AJC的介导破坏是通过骨膜肌动蛋白细胞骨架的拆卸来完成的，并且皮质素的下调 - 一种关键的肌动蛋白结合蛋白。 cortactin缺乏以前已与破坏血管内皮和肠上皮屏障的稳定。但是RSV-之间的因果关系尚未建立诱导的泄漏屏障，肌动蛋白细胞骨架重排和皮质素缺乏症。在此外，流行病学报告表明，暴露于环境特定物质之间有很强的联系（PM）和ALRI风险增加。纳米颗粒（NP）非常小PM，具有更大的能力与较小的颗粒相比，沉积在远端气道和逃避宿主防御措施中。我们的初步数据证明支气管上皮细胞对NP的暴露不仅增强了RSV诱导的AJC 拆卸和肌动蛋白细胞骨架破坏，但会增强病毒感染。根据我们的新颖观察，我们提出了中心假设，即A）RSV通过触发而引起气道上皮屏障的破坏沉积肌动蛋白细胞骨架，并通过下调cortactin； b）这种破坏纳米颗粒的上皮屏障会使RSV诱导的气道上皮损伤恶化。我们将测试我们的通过以下特定目的假设：目标1：确定依赖性肌动蛋白的作用 RSV诱导的气道上皮屏障功能障碍中的细丝动力学。使用人支气管上皮细胞我们将从小儿供体分离出和corortactin null小鼠的小鼠模型，我们将研究（i）RSV对肌动蛋白细胞骨架动力学，（ii）cortactin在肌动蛋白动力学和AJC结构上的功能作用，以及（iii） RAP-1的功能作用。我们还将使用详细详细介绍AJC效果的3-D人肺器官模型 RSV感染时的破坏，它提供了一个创新的平台来研究复杂的宿主环境互动。目标2：确定纳米颗粒是否增强了RSV诱导的气道上皮破坏障碍。使用体外和体内模型，我们将（i）表征粒径对屏障完整性的影响，（ii）研究氧化应激对AJC功能的作用，（iii）定义了暴露于NP对AJC的影响功能障碍。拟议的研究很重要，并且与NIH的使命相关，因为我们旨在探索 RSV感染对气道屏障完整性的临床相关后果以及如何暴露于环境污染物会恶化RSV感染。我们的方法具有创新性，因为它将提供新的机械洞察力 RSV在AJC拆卸中的作用，以及NP在增强RSV-的影响方面的新见解诱导AJC中断。确定的途径将为热干预提供新的目标，积极影响RSV疾病管理的潜力。