S-glutathionylation chemistry, glycolysis and obese allergic asthma

项目摘要 肥胖性哮喘是一个重要的临床问题，因为越来越多的患者受到影响 他们的疾病严重程度用现有的药物控制得很差。变化 代谢，以及氧化还原稳态的相关变化被认为有助于肥胖相关的 疾病该项目解决了S-谷胱甘肽化化学（PSSG，一种基于氧化还原的修饰， 涉及与谷胱甘肽结合的蛋白质半胱氨酸残基）有助于肥胖过敏性 哮喘我们已经发现了PSSG化学和小鼠肺中糖酵解之间的密切联系 与屋尘螨（HDM）诱导的过敏性气道疾病，并在原代鼻上皮细胞从 哮喘患者氧化还原蛋白质组学鉴定了糖酵解酶丙酮酸激酶M2（PKM 2）作为一种重要的糖酵解酶。 以S-谷胱甘肽化为靶标。PKM 2最近被认为与促炎性细胞因子的增加有关。 应答重要的是，PSSG和糖酵解在肥胖的情况下都增加，这表明PSSG 糖酵解的相关改变可能是肥胖过敏性哮喘的关键特征。这些发现使我们 假设糖酵解是促进过敏性气道疾病的关键过程，并且该过程 通过糖酵解酶PKM 2（PKM 2-SSG）的S-谷胱甘肽化增强。我们进一步假设， 与瘦型过敏性哮喘患者相比，这些过程在肥胖哮喘患者中增强。途径 抑制谷胱甘肽S-转移酶P（GSTP，S-谷胱甘肽化的催化剂）或促进去谷胱甘肽化 通过增加去谷胱甘肽化酶谷氧还蛋白-1（GLRX）的表达， 减弱糖酵解重编程和减少肥胖过敏性哮喘患者的气道疾病。在特定 目的#1：我们将确定GSTP的基因消除或抑制对小鼠糖酵解的影响 屋尘螨（HDM）-饮食诱导的肥胖（DIO）介导的肺病，以及来自 偏瘦和肥胖的过敏性哮喘患者。在具体目标#2中，我们建议评估GLRX调制的影响 在肺组织中，以减弱PSSG、PKM 2-SSG和糖酵解， 呼吸道疾病我们将利用转基因方法或将重组GLRX直接施用到患者的气道中。 存在过敏性气道疾病的瘦或肥胖小鼠。具体目标#3解决了丙酮酸的重要性 激酶M2用于瘦或肥胖过敏性气道疾病。我们将特异性地从气道中去除Pkm 2基因， 上皮细胞的小鼠，并提出药理学方法，以提高PKM 2活性的小鼠与现有的 HDM/DIO诱导的过敏性疾病和来自瘦或肥胖健康受试者的鼻上皮细胞，或 哮喘患者完成拟议的研究将提供新的机制的见解如何S- 谷胱甘肽化促进糖酵解重编程，并有可能提供临床/预防 针对S-谷胱甘肽化化学的相关策略，以减弱糖酵解， 肥胖过敏性哮喘患者的气道炎症和重塑。

PROJECT SUMMARY Obese asthma represents an important clinical problem due to the growing number of affected patients worldwide, and their disease severity which is poorly controlled with existing medications. Changes in metabolism, and associated changes in redox homeostasis are believed to contribute to obesity-associated diseases. This project addresses whether S-glutathionylation chemistry (PSSG, a redox-based modification of protein cysteine residues involving conjugation with glutathione) contributes to the pathogenesis of obese allergic asthma. We have discovered a close association between PSSG chemistry and glycolysis in lungs from mice with house dust mite (HDM)-induced allergic airways disease, and in primary nasal epithelial cells from asthmatics. Redox proteomics identified he glycolytic enzyme, pyruvate kinase M2 (PKM2) as an important target for S-glutathionylation. PKM2 had recently been implicated in the augmentation of pro-inflammatory responses. Importantly, both PSSG and glycolysis were increased in settings of obesity, suggesting that PSSG and linked alterations in glycolysis may be a key feature of obese allergic asthma. These findings led us to hypothesize that glycolysis is a critical process that promotes allergic airways disease, and that this process is enhanced by S-glutathionylation of the glycolytic enzyme PKM2 (PKM2-SSG). We furthermore hypothesize that these processes are augmented in obese asthmatic individuals compared to lean allergic asthmatics. Avenues to inhibit glutathione S-transferase P (GSTP, a catalyst of S-glutathionylation) or to promote de-glutathionylation of PKM2 by increasing expression of the de-glutathionylating enzyme, glutaredoxin-1 (GLRX), are anticipated to attenuate glycolytic reprogramming and to decrease airways disease in obese allergic asthmatics. In Specific Aim #1 we will determine the impact of genetic ablation or inhibition of GSTP in decreasing glycolysis in mice house dust mite (HDM)- diet induced obesity (DIO)- mediated lung disease, and in nasal epithelial cells from lean and obese allergic asthmatics. In Specific Aim #2 we propose to assess the impact of modulation of GLRX in lung tissues in order to attenuate PSSG, PKM2-SSG, and glycolysis in settings of lean or obese allergic airways disease. We will utilize transgenic approaches or administer recombinant GLRX directly into airways of lean or obese mice with existing allergic airways disease. Specific Aim #3 addresses the importance of pyruvate kinase M2 for lean or obese allergic airways disease. We will specifically ablate the Pkm2 gene from airway epithelial cells in mice and propose pharmacological approaches to enhance PKM2 activity in mice with existing HDM/DIO-induced allergic disease and in nasal epithelial cells derived from lean or obese healthy subjects or asthmatics. Completion of the proposed studies will provide new mechanistic insights into how S- glutathionylation facilitates glycolytic reprogramming and has the potential to offer clinically/translationally relevant strategies targeting S-glutathionylation chemistry in order to attenuate glycolysis, resultant lactate production, airways inflammation and remodeling in individuals with obese allergic asthma.