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：
10593065
负责人：
Venkataramana K Sidhaye
金额：
$ 60.29万
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/10593065
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 Cells Epithelium 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 Maps 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 Transcript United States Up-Regulation Variant airway epithelium airway remodeling alveolar destruction alveolar epithelium cigarette smoke cigarette smoke-induced epigenetic regulation exposure to cigarette smoke functional improvement improved 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.

项目摘要慢性阻塞性肺疾病（COPD）是美国肺气肿的第四大死亡原因进展和肺功能下降，表明粘膜校利清除率差和气道上皮变化的证据以及肺泡破坏。我们发现，在COPD中，粘附连接处有显着降低蛋白E-钙粘蛋白，它是决定细胞细胞粘附的主要成分，该发现已通过几名研究人员和大型队列研究了。在此提案中，我们将破译E-钙卫蛋白的影响调节气道和实质建模中上皮功能变化的调节，并确定是否 E-钙黏着蛋白增加可保护上皮和肺部免受香烟烟雾（CS）。我们提出了这一点肺上皮E-钙粘蛋白减少会导致上皮细胞的极性丧失，破坏上皮屏障F 并降低睫状节频率以促进气道功能障碍。此外，我们提出了E-的损失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AIM 1中，我们将确定E-Cadherin是否在细胞中敲低，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-钙粘着蛋白的丧失慢性烟雾暴露或COPD患者在COPD的发展中起因果作用。具体而言，我们将确定肺E-钙粘蛋白损失是否改变上皮极性并降低纤毛的比例空气表面液体的频率，粘膜清除和维护。此外，我们将确定是否e- 钙粘蛋白损失使用小鼠模型促进实质重塑和炎症。在AIM 2中，我们将研究在CS暴露和COPD的背景下，E-钙粘蛋白减少的新型机制。具体而言，我们将研究启动子的表观遗传调节是否有助于mRNA和蛋白质的降低丰富。我们的数据表明，CDH1在富含调节元素的区域中被甲基化，因此我们将研究这些元素以确定CS暴露发生的变化。此外，我们已经确定了转化修饰与人群研究中的电子钙粘蛋白功能相关。具体来说，我们会的确定E-钙粘着蛋白的末端诱导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鉴定的多态性或单倍型遗传上将其纳入肺细胞系中，以确定这些是否影响基于电子钙粘蛋白的细胞 - 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对CS的敏感性增加接触。在AIM 3中，我们将执行概念研究证明，以确定E-钙黏着蛋白的上调是否作为原代人细胞和小鼠模型中的治疗策略。此外，我们将确定是否操纵AIM 2中确定的途径上调上调E-钙粘蛋白并在CS背景下提高功能接触。