Parkin in mitochondrial dysfunction and airway inflammation of obese asthma

Parkin 在肥胖哮喘线粒体功能障碍和气道炎症中的作用

基本信息

批准号：
10264924
负责人：
Hong W Chu
金额：
$ 73.14万
依托单位：
NATIONAL JEWISH HEALTH
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：
2020-09-16 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10264924
关键词：
Address Adrenal Cortex Hormones Allergens Arginine Asthma Attenuated Basic Science Biological Availability Bronchoalveolar Lavage Fluid CCL26 gene CRISPR/Cas technology Cell Culture Techniques Clinical Disease Eotaxin Epithelial Cells Equilibrium Exposure to Gene Activation Goals High Fat Diet Human Inflammation Inhalation Interferon Type II Interferons Interleukin-13 Knock-out Liquid substance Lung Measures Mediating Mitochondria Mitochondrial DNA Mus N,N-dimethylarginine Neutrophil Infiltration Nitric Oxide Nitric Oxide Synthase Obesity PINK1 gene Palmitic Acids Pathogenesis Production Public Health Reactive Oxygen Species Research Research Personnel Respiration Role STAT1 gene STAT6 gene Sampling Saturated Fatty Acids Scientist Serum Signal Pathway Signal Transduction Slice Stimulator of Interferon Genes TBK1 gene Testing Thinness Transcription Repressor Translational Research Up-Regulation airway epithelium airway inflammation asthmatic asthmatic airway bronchial epithelium chemokine cytokine eosinophil eosinophilic inflammation experience inhibitor/antagonist innovation interleukin-13 receptor mitochondrial dysfunction mouse model multidisciplinary neutrophil nitrosative stress novel parkin gene/protein response therapeutic evaluation ubiquitin-protein ligase

项目摘要

Project Summary/Abstract The goal of this R01 application is to determine the mechanisms by which mitochondrial dysfunction in obese asthma exacerbates airway inflammation. Nearly 40% of asthmatics in the U.S. are obese. Obese (vs. lean) asthmatics experience more frequent exacerbations, poorer response to inhaled corticosteroids and worse asthma control. There is an unmet need to study the novel pathogenesis of obese asthma. We have found mitochondrial dysfunction in obese asthma airway epithelium, including increased respiration, glycolytic rates, reactive oxygen species, and greater numbers of dysfunctional mitochondria. We discovered up-regulation of Parkin (Park2) in obese asthma airway epithelium. Parkin is an E3 ubiquitin ligase that regulates mitophagy by degrading defective mitochondria. We further observed increased levels of mitochondrial DNA (mtDNA) and palmitic acid (PA) levels in obese asthma bronchoalveolar lavage fluid and serum samples. Moreover, type 2 cytokine IL-13, type 1 cytokine IFN-g and palmitic acid (PA, a saturated fatty acid increased in obese asthma) increased airway epithelial Parkin levels. Parkin is essential to IL-13-mediated mtDNA release and airway neutrophilic and eosinophilic inflammation in mice and in human airway epithelial cells. We hypothesize that Parkin is up-regulated in obese asthma airways with dysfunctional mitochondria, which is amplified by the type 1 or type 2 cytokine milieu, enhancing mitochondrial DNA release and exaggerating airway inflammation. In Aim 1, we will determine the mechanisms by which Parkin is up-regulated in obese asthma by measuring Parkin, PA, mitophagy, type 1 and type 2 cytokines, and nitric oxide (NO), and performing human airway epithelial cell and precision-cut lung slice cultures to determine if IFN-g, IL-13 and PA increase Parkin in part by reducing transcriptional repressor THAP11. We then test if restoring NO bioavailability reduces Parkin activity. In Aim 2, we will determine how Parkin enhances airway mtDNA release and neutrophilic inflammation. By using Parkin, TLR9 and STAT1 knockout human airway epithelial culture and mouse models, we will test if excessive Parkin in a type 1 cytokine setting of obese asthma promotes mtDNA release, and amplifies neutrophilic inflammation through the TLR9/STAT1 signaling axis. In Aim 3, we will determine the mechanisms by which Parkin enhances airway eosinophilic inflammation. We will test if Parkin-mediated mtDNA release in a type 2 cytokine setting amplifies STAT6 signaling and pro-eosinophilic cytokine (e.g. eotaxin) production. Research findings from our proposed studies will likely provide several key targets (e.g. Parkin, TLR9) to treat obese asthma, a heterogenous disease currently without specific therapies.

项目概要/摘要 R01 应用的目标是确定肥胖者线粒体功能障碍的机制哮喘会加剧气道炎症。在美国，近 40% 的哮喘患者患有肥胖症。肥胖（与瘦）哮喘患者会经历更频繁的恶化、对吸入皮质类固醇的反应更差甚至更糟哮喘控制。研究肥胖哮喘的新发病机制的需求尚未得到满足。我们发现肥胖哮喘气道上皮线粒体功能障碍，包括呼吸增加、糖酵解速率增加、活性氧和大量功能失调的线粒体。我们发现上调肥胖哮喘气道上皮细胞中的 Parkin (Park2)。 Parkin 是一种 E3 泛素连接酶，通过以下方式调节线粒体自噬：降解有缺陷的线粒体。我们进一步观察到线粒体 DNA (mtDNA) 水平增加肥胖哮喘支气管肺泡灌洗液和血清样本中的棕榈酸（PA）水平。此外，类型 2 细胞因子 IL-13、1 型细胞因子 IFN-g 和棕榈酸（PA，一种在肥胖哮喘中增加的饱和脂肪酸）气道上皮 Parkin 水平升高。 Parkin 对于 IL-13 介导的 mtDNA 释放和气道至关重要小鼠和人气道上皮细胞的中性粒细胞和嗜酸性粒细胞炎症。我们假设 Parkin 在线粒体功能障碍的肥胖哮喘气道中上调，并通过以下因素放大： 1 型或 2 型细胞因子环境，增强线粒体 DNA 释放并扩大气道炎。在目标 1 中，我们将通过以下方法确定 Parkin 在肥胖哮喘中上调的机制：测量 Parkin、PA、线粒体自噬、1 型和 2 型细胞因子以及一氧化氮 (NO)，并执行人类气道上皮细胞和精密切割肺切片培养物以确定 IFN-g、IL-13 和 PA 是否会增加 Parkin 部分通过减少转录阻遏蛋白 THAP11 来实现。然后我们测试恢复 NO 生物利用度是否会降低 Parkin 活动。在目标 2 中，我们将确定 Parkin 如何增强气道 mtDNA 释放和中性粒细胞炎症。通过使用 Parkin、TLR9 和 STAT1 敲除的人气道上皮培养物和小鼠模型，我们将测试是否肥胖哮喘 1 型细胞因子环境中过量 Parkin 促进 mtDNA 释放，并放大通过 TLR9/STAT1 信号轴产生中性粒细胞炎症。在目标 3 中，我们将确定机制 Parkin 通过这种方式增强气道嗜酸性粒细胞炎症。我们将测试 Parkin 介导的 mtDNA 释放是否在 2 型细胞因子设置可放大 STAT6 信号传导和促嗜酸性细胞因子（例如嗜酸细胞趋化因子）的产生。我们拟议的研究结果可能会提供几个关键靶标（例如 Parkin、TLR9）来治疗肥胖哮喘是一种异质性疾病，目前尚无特效疗法。