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

• 批准号: 10457989
• 负责人: Hong W Chu
• 金额: $ 72.92万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 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/10457989
• 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; Parkin; 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; antagonist; asthmatic; asthmatic airway; bronchial epithelium; chemokine; cytokine; eosinophil; eosinophilic inflammation; experience; inhibitor; innovation; interleukin-13 receptor; mitochondrial dysfunction; mouse model; multidisciplinary; neutrophil; nitrosative stress; novel; obesity-associated asthma; response; therapeutic evaluation; ubiquitin-protein ligase

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.

项目总结/摘要 R 01应用的目的是确定肥胖患者线粒体功能障碍的机制。 哮喘加剧了气道炎症。美国近40%的哮喘患者肥胖。肥胖（vs.瘦） 哮喘患者经历更频繁的恶化，对吸入性皮质类固醇的反应更差， 哮喘控制研究肥胖性哮喘的新发病机制的需求尚未得到满足。我们发现 肥胖哮喘气道上皮细胞的线粒体功能障碍，包括呼吸增加，糖酵解速率， 活性氧和大量功能失调的线粒体。我们发现， Parkin（Park 2）在肥胖哮喘气道上皮中的表达。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）和 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，肥胖性哮喘中饱和脂肪酸增加） 增加气道上皮帕金水平。Parkin对IL-13介导的mtDNA释放和气道炎症至关重要 小鼠和人气道上皮细胞中的嗜酸性粒细胞和嗜酸性粒细胞炎症。我们假设 Parkin在伴有线粒体功能障碍的肥胖哮喘气道中上调， 1型或2型细胞因子环境，增强线粒体DNA释放并扩大气道 炎症在目标1中，我们将通过以下方式确定肥胖哮喘中Parkin上调的机制： 测量Parkin、PA、线粒体自噬、1型和2型细胞因子和一氧化氮（NO），并进行人 气道上皮细胞和精确切割的肺切片培养，以确定IFN-g，IL-13和PA是否增加Parkin， 部分通过减少转录抑制因子THAP 11。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，我们将确定帕金如何增强气道mtDNA释放和嗜酸性炎症。 通过使用Parkin、TLR 9和STAT 1敲除的人气道上皮培养物和小鼠模型，我们将测试是否 在肥胖性哮喘的1型细胞因子环境中，过度的Parkin促进mtDNA的释放， 通过TLR 9/STAT 1信号传导轴的嗜酸性炎症。在目标3中，我们将确定 帕金通过其增强气道嗜酸性炎症。我们将测试帕金森介导的线粒体DNA释放是否在一个 2型细胞因子设置放大了STAT 6信号传导和嗜酸性细胞因子（例如嗜酸细胞活化趋化因子）的产生。 我们提出的研究的研究结果可能会提供几个关键的治疗靶点（例如Parkin，TLR 9）。 肥胖性哮喘是一种异质性疾病，目前尚无特异性治疗方法。