The Unfolded Protein Response in Human Airway and Alveolar Epithelial Cells

人类气道和肺泡上皮细胞中未折叠的蛋白质反应

基本信息

批准号：
7837633
负责人：
SALIM MERALI
金额：
$ 7.5万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-11 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7837633
关键词：
Alveolar Anti-Inflammatory Agents Anti-inflammatory Antioxidants Apoptosis Apoptotic Biochemical Pathway Biological Calnexin Cell Death Cells Chronic Chronic Obstructive Airway Disease Cigarette Cigarette Smoker Data Development Disease Endoplasmic Reticulum Epithelial Cells Erythroid Fibrosis GRP78 gene Gene Expression Gold Harvest Homeostasis Human In Situ In Vitro Individual Lasers Lectin Lung Lung Inflammation Lung diseases Metaplasia Microscopy Molecular Chaperones NF-E2-related factor 2 Nuclear Oxidants Pathogenesis Phosphotransferases Predisposition Protein Disulfide Isomerase Proteins Pulmonary Emphysema Research Risk Factors Signal Pathway Signal Transduction Smoke Smoker Staging Stream Structure of parenchyma of lung Techniques Testing Tissue Sample Tissues Up-Regulation Work abstracting activating transcription factor 4 base calreticulin cell type cigarette smoke-induced cigarette smoking cigarette smoking common cellular transcription factor ATF endoplasmic reticulum stress improved novel oxidant stress prevent protein kinase R public health relevance response transcription factor transcription factor CHOP

项目摘要

DESCRIPTION (provided by applicant): Cigarette smoking induced chronic obstructive pulmonary disease (COPD) is characterized by alveolar epithelial cell death and airway metaplasia and fibrosis. We have shown that the lungs of chronic cigarette smokers without COPD manifest a compensatory response to endoplasmic reticulum stress termed the unfolded protein response (UPR) and that the UPR-related transcription factors, ATF-4 and Nrf2, proteins which regulate anti-oxidant gene expression, are up-regulated by CS in cultured airway epithelial cells. These data raise the possibility that the UPR protects lung cells against oxidant stress and the development of COPD. However, when ER stress is sufficiently severe and protein homeostasis cannot be restored, the UPR causes apoptosis by up-regulating expression of the pro-apoptotic transcription factor, CHOP. These latter observations raise the alternative possibility that the UPR may contribute to the pathogenesis of emphysema by inducing cell apoptosis. Our previous studies were performed in whole lung lysates and cultured airway epithelial cells. We have not studied the in situ response of specific lung cell types to cigarette smoke exposure. Accordingly, the specific Aim of this 2 year project is to characterize the UPR induced by cigarette smoke in a lung cell involved in the pathogenesis of COPD, i.e., airway epithelial cells. We will test the hypotheses that UPR response to cigarette smoke is impaired in the airway epithelial cells of subjects with COPD. Expression of the UPR hallmark proteins (i.e., GRP78, calreticulin, calnexin and protein disulfide isomerase), oxidant defense (i.e., Nrf2 and ATF4) and apoptosis (i.e., CHOP) will be compared in airway epithelial cells from subjects with and without COPD. Cells will be harvested from ex-- smokers without COPD (n=10) and ex-smokers with COPD in GOLD stages II (n=10) and IV (n=10) using the technique of laser capture microscopy applied to HOPE fixed, inflated lung tissue sections. A better understanding of the cellular basis of the UPR induced by CS should enhance our understanding of the pathogenesis of COPD. PUBLIC HEALTH RELEVANCE: Cigarette smoking is a risk factor for chronic obstructive pulmonary disease (COPD), a disease characterized by inflammation, and lung cell death. However, the pathogenesis of this disease remains incompletely understood and current therapies are inadequate. Our recent work shows that a novel biochemical pathway induced in the lung by cigarette smoking can be protective. This response and its biological consequences will be studied in cell type known to be involved with COPD. The results obtained are expected to improve our understanding of cigarette smoke-related lung disease and contribute to the development of new and better treatments. (End of Abstract)

描述（由申请人提供）：吸烟引起的慢性阻塞性肺疾病（COPD）的特征是肺泡上皮细胞死亡和气道变质和纤维化。 We have shown that the lungs of chronic cigarette smokers without COPD manifest a compensatory response to endoplasmic reticulum stress termed the unfolded protein response (UPR) and that the UPR-related transcription factors, ATF-4 and Nrf2, proteins which regulate anti-oxidant gene expression, are up-regulated by CS in cultured airway epithelial cells.这些数据增加了UPR保护肺细胞免受氧化应激和COPD发展的可能性。但是，当ER应激足够严重并且无法恢复蛋白质稳态时，UPR通过上调促凋亡转录因子的表达而引起凋亡。这些后一种观察结果提出了一种替代可能性，即UPR可能通过诱导细胞凋亡来促进肺气肿的发病机理。我们先前的研究是在整个肺裂解物和培养的气道上皮细胞中进行的。我们尚未研究特定的肺细胞类型对香烟烟雾暴露的原位反应。因此，该2年项目的具体目的是表征与COPD发病机理（即气道上皮细胞）中的肺部烟雾引起的UPR。我们将测试以COPD受试者的气道上皮细胞中UPR对香烟烟雾的反应的假设。 UPR Hallmark蛋白（即GRP78，钙网蛋白，钙网蛋白和蛋白质二硫化物异构酶），氧化剂防御（即NRF2和ATF4）和凋亡（即CHOP）的表达将在患有和没有COPD的受试者的受试者中进行比较。使用激光捕获显微镜的技术，将从没有COPD的EX-吸烟者（n = 10）和具有COPD的Ex-Smoker中收集细胞，并使用激光捕获显微镜的技术来收获细胞。对CS诱导的UPR的细胞基础的更好理解应增强我们对COPD发病机理的理解。公共卫生相关性：吸烟是慢性阻塞性肺疾病（COPD）的危险因素，该疾病是炎症和肺部细胞死亡的特征。然而，该疾病的发病机理仍未完全理解，目前的疗法不足。我们最近的工作表明，通过吸烟在肺部引起的一种新型的生化途径可以保护。该反应及其生物学后果将在已知与COPD有关的细胞类型中进行研究。预计获得的结果将提高我们对香烟烟有关的肺部疾病的理解，并有助于新的和更好的治疗方法的发展。（抽象的结尾）