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DJ-1 pathway impairment in alveolar type II cells in emphysema.

肺气肿中肺泡 II 型细胞的 DJ-1 通路损伤。

基本信息

批准号：
9108997
负责人：
Beata Kosmider
金额：
$ 39万
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：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9108997
关键词：
Address Adenoviruses Alveolar Alveolar wall Antioxidants Applications Grants Binding Cells Clinical Cysteine Cytoprotection Data Development Disease Disease Progression Epithelial Cells Exposure to Foundations Future Human Impairment In Vitro Injury Knockout Mice Lead Lung diseases Lung volume reduction surgery Mass Spectrum Analysis Methionine Methods Modification Mus Oxidation-Reduction Oxidative Stress Oxides Ozone Pathogenicity Pathway interactions Patients Pattern Pharmacology Positioning Attribute Post-Translational Protein Processing Proteins Pulmonary Emphysema Reactive Oxygen Species Reporting Role Smoker Structure of parenchyma of lung Sulfinic Acids Sulfonic Acids System Testing Time Ubiquitination Wild Type Mouse alveolar destruction alveolar type II cell biobank cell injury cigarette smoke-induced cigarette smoking cigarette smoking effective therapy experimental study in vivo influenzavirus mutant novel novel therapeutics nuclear factor-erythroid 2 oxidation public health relevance sensor

项目摘要

DESCRIPTION (provided by applicant): Emphysema consists of a unique pattern of alveolar wall destruction and there is limited therapy against this disease. Cigarette smoking induces oxidative stress and is the most common cause of pulmonary emphysema. Our application seeks to define the mechanism of DJ-1 pathway impairment in this disease. DJ-1 is a multifunctional protein that protects cells from oxidative stress. This depends on posttranslational modifications of cysteine and methionine residues within DJ-1. Kelch-like ECH-associated protein 1 (KEAP1) and CR6-interacting Factor 1 (CRIF1) have been also proposed as factors, which regulate oxidative stress. DJ-1-CRIF1-KEAP1 interaction induces antioxidant defense systems against alveolar epithelial cell injury by cigarette smoke. We propose to study the mechanism of DJ-1 cytoprotective function against human primary alveolar epithelial cell injury by cigarette smoke in vitro and ex vivo. We will determine oxidation of cysteine and methionine residues within DJ-1 using mass spectrometry analysis. We will use mutant constructs to identify, which oxidizable cysteine and methionine residue(s) are critical for DJ-1 function in protecting against injury induced by cigarette smoke. We will focus on cysteine residue at position 106 (Cys-106), which is a sensor of oxidative stress within DJ-1. To further determine the DJ-1 pathway, we will analyze DJ-1, CRIF1 and KEAP1 interacting domains using deletion constructs. We will also use in our studies alveolar type II cells isolated from patients with mild, moderate and severe emphysema. This unique approach will allow us to identify the mechanism of DJ-1 pathway impairment in emphysema progression. We will validate our results in alveolar type II cells in vivo. Wild-type mice will be exposed to cigarette smoke to determine DJ-1 cysteine and methionine modifications by mass spectrometry under oxidative stress conditions. We will also determine whether adenovirus DJ-1 will rescue of alveolar type II cell injury due to cigarette smoke in DJ-1 knockout mice. Study of the mechanism of the DJ-1 pathway impairment in emphysema may lead to the development of novel pharmacological strategies to slow the progression of this disease.

描述（由申请人提供）：肺气肿由一种独特的肺泡壁破坏模式组成，对这种疾病的治疗有限。吸烟引起氧化应激，是肺气肿最常见的原因。我们的申请旨在确定这种疾病中DJ-1通路受损的机制。DJ-1是一种多功能蛋白质，可保护细胞免受氧化应激。这取决于DJ-1内半胱氨酸和甲硫氨酸残基的翻译后修饰。Kelch样ECH相关蛋白1（KEAP 1）和CR 6相互作用因子1（CRIF 1）也被认为是调节氧化应激的因子。DJ-1-CRIF 1-KEAP 1相互作用诱导抗氧化防御系统对抗香烟烟雾引起的肺泡上皮细胞损伤。我们拟在体外和离体研究DJ-1对香烟烟雾所致人原代肺泡上皮细胞损伤的细胞保护作用机制。我们将使用质谱分析确定DJ-1中半胱氨酸和蛋氨酸残基的氧化。我们将使用突变体构建体来鉴定哪些可氧化的半胱氨酸和甲硫氨酸残基对于DJ-1在保护免受香烟烟雾诱导的损伤中的功能是关键的。我们将专注于106位半胱氨酸残基（Cys-106），这是DJ-1内氧化应激的传感器。为了进一步确定DJ-1通路，我们将使用缺失构建体分析DJ-1、CRIF 1和KEAP 1相互作用结构域。我们还将在我们的研究中使用从轻度、中度和重度肺气肿患者中分离的肺泡II型细胞。这种独特的方法将使我们能够确定肺气肿进展中DJ-1通路受损的机制。我们将在肺泡II型细胞体内验证我们的结果。野生型小鼠将暴露于香烟吸烟以在氧化应激条件下通过质谱法测定DJ-1半胱氨酸和甲硫氨酸修饰。我们还将确定腺病毒DJ-1是否会拯救肺泡II型细胞损伤由于香烟烟雾在DJ-1基因敲除小鼠。研究肺气肿中DJ-1通路受损的机制可能会导致开发新的药理学策略来减缓这种疾病的进展。