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

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

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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.

项目摘要/摘要该R01应用的目的是确定肥胖中线粒体功能障碍的机制哮喘加剧了气道炎症。在美国，近40％的哮喘患者肥胖。肥胖（vs.精益）哮喘患者经历更频繁的加重，对吸入皮质类固醇的反应较差，并且更差哮喘控制。研究了肥胖哮喘的新型发病机理的需求未满足。我们找到了肥胖哮喘气道上皮的线粒体功能障碍，包括呼吸升高，糖酵解速率，活性氧和更多功能障碍线粒体。我们发现了上调肥胖哮喘气道上皮的Parkin（Park2）。帕金是一种E3泛素连接酶，可调节线粒体有缺陷的线粒体降解。我们进一步观察到线粒体DNA（mtDNA）和肥胖的哮喘支气管肺泡液和血清样品中的棕榈酸（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，肥胖哮喘的饱和脂肪酸增加）呼吸道上皮帕金水平提高。 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中的嗜中性粒细胞和嗜酸性炎症。我们假设这一点帕金在肥胖的哮喘气道中被上调，线粒体功能失调，这被放大 1型或2型细胞因子环境，增强线粒体DNA释放并夸大气道炎。在AIM 1中，我们将确定Parkin在肥胖哮喘中上调的机制测量帕金，宾夕法尼亚州，线粒体，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是否增加了Parkin 部分通过减少转录阻遏物THAP11。然后，我们测试是否恢复无生物利用度会减少帕金活动。在AIM 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敲除人类气道上皮文化和老鼠模型，我们将测试是否是否肥胖哮喘的1型细胞因子环境中过多的parkin促进mtDNA释放，并放大通过TLR9/STAT1信号轴的中性粒细胞炎症。在AIM 3中，我们将确定机制帕金通过此增强气道嗜酸性炎症。我们将测试Parkin介导的mtDNA是否在A中释放 2型细胞因子设置放大了STAT6信号传导和促嗜酸性细胞因子（例如Eotaxin）的产生。我们提出的研究的研究结果可能会提供几个关键目标（例如Parkin，TLR9）来治疗肥胖哮喘，一种目前没有特定疗法的异质疾病。