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: 10675493
• 负责人: Hong W Chu
• 金额: $ 71.84万
• 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/10675493
• 关键词: Address; Adrenal Cortex Hormones; Allergens; Arginine; Asthma; 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; asthma exacerbation; 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.

项目摘要/摘要 此R01应用程序的目标是确定肥胖患者线粒体功能障碍的机制 哮喘会加重呼吸道炎症。在美国，近40%的哮喘患者患有肥胖症。肥胖(VS瘦) 哮喘患者的病情加重更频繁，对吸入皮质类固醇的反应更差，情况更糟 哮喘控制。研究肥胖哮喘的新发病机制是一个尚未得到满足的需求。我们发现了 肥胖哮喘呼吸道上皮线粒体功能障碍，包括呼吸增加，糖酵解率， 活性氧物种，以及更多功能失调的线粒体。我们发现上调了 Parkin(Park2)在肥胖哮喘呼吸道上皮中的表达。Parkin是一种E3泛素连接酶，通过 降解有缺陷的线粒体。我们进一步观察到线粒体DNA(MtDNA)水平和 肥胖性哮喘患者支气管肺泡灌洗液和血清中棕榈酸的水平。此外，类型2 细胞因子IL-13、1型细胞因子干扰素-g和棕榈酸(PA，一种饱和脂肪酸在肥胖性哮喘中升高) 呼吸道上皮帕金素水平升高。Parkin在IL-13介导的线粒体DNA释放和呼吸道中起重要作用 小鼠和人类呼吸道上皮细胞中的中性粒细胞和嗜酸性炎症。我们假设 Parkin在线粒体功能障碍的肥胖哮喘呼吸道中上调，线粒体功能障碍通过 1型或2型细胞因子环境，促进线粒体DNA释放，夸大呼吸道 发炎。在目标1中，我们将确定Parkin在肥胖性哮喘中上调的机制 测量Parkin、PA、有丝分裂、1型和2型细胞因子以及一氧化氮(NO)，并进行人类 呼吸道上皮细胞和肺切片培养检测干扰素-g、白介素13和纤溶酶原激活剂是否增加肺组织Parkin 部分通过减少转录抑制因子THAP11。然后，我们测试恢复无生物利用度是否可以减少停车 活动。在目标2中，我们将确定Parkin如何促进气道mtDNA释放和中性粒细胞炎症。 通过使用Parkin、TLR9和STAT1基因敲除的人呼吸道上皮细胞培养和小鼠模型，我们将测试 肥胖哮喘1型细胞因子环境中过量的Parkin促进mtDNA释放，并放大 中性粒细胞炎症通过TLR9/STAT1信号轴。在目标3中，我们将确定机制 帕金通过这种方式增强呼吸道嗜酸性炎症。我们将测试帕金介导的线粒体DNA是否在 2型细胞因子的设置放大了STAT6信号和嗜酸性细胞因子(如嗜酸性粒细胞因子)的产生。 我们拟议研究的研究结果可能会提供几个关键的治疗靶点(例如Parkin，TLR9) 肥胖性哮喘是一种异质性疾病，目前还没有特殊的治疗方法。