Role of E-cadherin in modulating airway epithelial function and parenchymal remodeling

E-钙粘蛋白在调节气道上皮功能和实质重塑中的作用

基本信息

批准号：
10374001
负责人：
Venkataramana K Sidhaye
金额：
$ 61.45万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-05 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10374001
关键词：
Adherens Junction Adhesions Affect Air Area Binding CDH1 gene Cause of Death Cell Line Cell model Cell physiology Cell-Cell Adhesion Cells ChIP-seq Characteristics Chronic Chronic Bronchitis Chronic Obstructive Pulmonary Disease Cohort Studies DNA Methylation DNA Methylation Inhibition DNA methylation profiling Data Development Disease Down-Regulation E-Cadherin Elements Environment Epigenetic Process Epithelial Epithelial Cells Exposure to Fibrosis Frequencies Functional disorder Genes Genetic Engineering Genetic Polymorphism Haplotypes Human In Vitro Inflammation Inhalation Injury Knowledge Lead Liquid substance Lung Lung diseases Maintenance Measures Messenger RNA Methylation Modeling Mucociliary Clearance Mus Nuclear Pathway interactions Patients Phenotype Population Study Post-Translational Protein Processing Predisposition Proteins Pulmonary Emphysema Quantitative Trait Loci Regulation Regulatory Element Research Personnel Risk Role Secretor blood group alpha-2-fucosyltransferase Site Smoke Surface Testing Therapeutic Tissues Trachea United States Up-Regulation Variant airway epithelium airway remodeling alveolar destruction alveolar epithelium base cigarette smoke cigarette smoke-induced epigenetic regulation exposure to cigarette smoke improved improved functioning knock-down methylation pattern morphometry mortality mouse model novel overexpression preservation prevent promoter protein E pulmonary function decline restoration

项目摘要

PROJECT SUMMARY Chronic Obstructive Pulmonary Disease (COPD) is the 4th leading cause of death in the US with emphysema progression and lung function decline with evidence of poor mucociliary clearance and airway epithelial changes as well as alveolar destruction. We have found that in COPD there is a significant decrease in adherens junction protein E-cadherin, which is a primary component dictating cell-cell adhesion a finding which has been verified by several investigators and in large cohort studied. In this proposal, we will decipher the effects of E-cadherin regulation in promoting changes in epithelial function in the airways and parenchymal modeling and determine if increased E-cadherin protects the epithelia and the lungs from cigarette smoke (CS). We propose that decreased lung epithelial E-cadherin leads to loss of polarity of the epithelial cell, disrupts the epithelial barrier f and decreases ciliary beat frequency to promote airway dysfunction. Furthermore, we propose that loss of E- cadherin promotes parenchymal remodeling. We will dissect two specific mechanisms which could contribute to decreased protein levels. In Aim 1, we will determine if E-cadherin knockdown in cells, ex vivo trachea and mouse models lead to epithelial dysfunction and altered tissue integrity indicating that loss of E-cadherin with chronic smoke exposure or in patients with COPD serves a causal role in the development of COPD. Specifically, we will determine if lung E-cadherin loss alters epithelial polarity and decreases ciliary beat frequency, mucociliary clearance, and maintenance of the air surface liquid. In addition, we will determine if E- cadherin loss promotes parenchymal remodeling and inflammation using mouse models. In Aim 2, we will study novel mechanisms by which E-cadherin is decreased in the context of CS exposure and COPD. Specifically, we will study if epigenetic regulation of the promoter contributes to decrease in mRNA and protein abundance. Our data shows that CDH1 is methylated in an area rich with regulatory elements, so we will study these elements to determine what is altered with CS exposure. In addition, we have identified a post- translational modification which correlates with E-cadherin function in population studies. Specifically, we will determine if terminal fucosylation of E-cadherin increases its adhesion strength to enhance function. We have identified a specific identified polymorphism or a haplotype that is associated with functional changes and will genetically engineer this into a lung cell line to determine if these affect E-cadherin based cell-cell adhesion and function. In addition, we will determine if mice lacking this fucosylation have increased susceptibility to CS exposure. In Aim 3, we will perform proof of concept studies to determine if upregulation of E-cadherin serves as a therapeutic strategy in primary human cells and mouse models. In addition, we will determine if manipulating the pathways identified in Aim 2 upregulate E-cadherin and improve function in the context of CS exposure.

项目概要慢性阻塞性肺疾病 (COPD) 是美国第四大肺气肿死亡原因进展和肺功能下降，并有粘液纤毛清除不良和气道上皮变化的证据以及肺泡破坏。我们发现，在慢性阻塞性肺病中，粘附连接显着减少蛋白质 E-钙粘蛋白，它是决定细胞间粘附的主要成分，这一发现已得到证实几位研究人员在大型队列中进行了研究。在本提案中，我们将破译 E-钙粘蛋白的作用调节促进气道上皮功能和实质建模的变化，并确定是否增加的 E-钙粘蛋白可以保护上皮细胞和肺部免受香烟烟雾 (CS) 的影响。我们建议肺上皮E-钙粘蛋白减少导致上皮细胞极性丧失，破坏上皮屏障并降低纤毛跳动频率以促进气道功能障碍。此外，我们建议失去 E- 钙粘蛋白促进实质重塑。我们将剖析两种可能有助于蛋白质水平降低。在目标 1 中，我们将确定细胞、离体气管和小鼠模型导致上皮功能障碍和组织完整性改变，表明 E-钙粘蛋白的丢失长期接触烟雾或患有慢性阻塞性肺病的患者在慢性阻塞性肺病的发展中起着因果作用。具体来说，我们将确定肺 E-钙粘蛋白损失是否会改变上皮极性并减少纤毛跳动频率、粘液纤毛清除和空气表面液体的维持。此外，我们将确定是否 E- 使用小鼠模型，钙粘蛋白损失促进实质重塑和炎症。在目标 2 中，我们将研究在 CS 暴露和 COPD 背景下 E-钙粘蛋白减少的新机制。具体来说，我们将研究启动子的表观遗传调控是否会导致 mRNA 和蛋白质的减少丰富。我们的数据显示CDH1在富含调控元件的区域被甲基化，因此我们将研究这些元素来确定 CS 暴露会改变什么。此外，我们还确定了一个后群体研究中与 E-钙粘蛋白功能相关的翻译修饰。具体来说，我们将确定 E-钙粘蛋白的末端岩藻糖基化是否会增加其粘附强度以增强功能。我们有确定了与功能变化相关的特定多态性或单倍型，并将将其基因工程化到肺细胞系中，以确定它们是否影响基于 E-钙粘蛋白的细胞粘附和功能。此外，我们将确定缺乏这种岩藻糖基化的小鼠是否会增加对 CS 的易感性接触。在目标 3 中，我们将进行概念验证研究，以确定 E-钙粘蛋白的上调是否作为原代人类细胞和小鼠模型的治疗策略。此外，我们将确定是否操纵 Aim 2 中确定的途径上调 E-钙粘蛋白并改善 CS 中的功能接触。