S-nitrosylation signaling in asthma

哮喘中的 S-亚硝基化信号传导

基本信息

批准号：
10662243
负责人：
JONATHAN S. STAMLER
金额：
$ 40.25万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10662243
关键词：
ADRBK1 gene Acute Adrenergic Agents Affect Agonist Air Movements American Arrestin Beta 1 Asthma Biochemistry Bronchodilator Agents Cardiac Cell surface Cells Chronic Clinical Collaborations Complement Complex Coupled Cysteine Data Development Diagnosis Drug Receptors Enzymes Epithelial Cells Event Excision Exhibits Functional disorder G protein coupled receptor kinase G-Protein-Coupled Receptors GTP-Binding Proteins Genetic Glutathione Glutathione Reductase Height Heterotrimeric GTP-Binding Proteins Homeostasis Human Inhalation Injury Intervention Kinetics Knock-in Link Lung Maintenance Mediating Mediator Modeling Molecular Molecular Weight Mus Mutant Strains Mice Mutate Mutation Nitric Oxide Nitric Oxide Synthase Oxidation-Reduction Oxidoreductase Pathogenesis Pathology Pathway interactions Patient Selection Patients Pharmaceutical Preparations Phosphorylation Physiology Point Mutation Positioning Attribute Post-Translational Protein Processing Proteins Publishing Pulmonary Function Test/Forced Expiratory Volume 1 Receptor Signaling Reduced Glutathione Regulation Resistance Role S-Nitrosoglutathione S-Nitrosothiols Sampling Signal Pathway Signal Transduction Single Nucleotide Polymorphism Site Smooth Muscle Myocytes Sulfhydryl Compounds System Tachyphylaxis Testing Work airway hyperresponsiveness airway inflammation airway remodeling arrestin 2 asthma model asthmatic asthmatic patient bench to bedside beta-2 Adrenergic Receptors beta-arrestin biological systems clinically relevant cohort cost desensitization efficacy testing improved inhibitor innovation mortality mouse model patient subsets personalized approach pre-clinical preservation prevent programs protective effect protein function pulmonary function receptor receptor function synergism

项目摘要

PROJECT SUMMARY/ABSTRACT Project 1 Asthma afflicts over 7% of Americans and is the result of chronic inflammation of the airways leading to airway remodeling and hyperresponsiveness that impedes air flow. Severe asthma is asthma that remains problematic despite maximal intervention with conventional asthma therapies, and it accounts for the majority of the mortality and cost of asthma. Dysfunction of airway β2-adrenergic receptor (β2AR) signaling contributes to severe asthma pathogenesis, but the precise signaling mechanisms responsible are unclear. While activation of β2AR using inhaled “β-agonist” drugs is a mainstay of acute asthma treatment, overactivation of β2AR is detrimental and can be fatal. Airway β2ARs signal both through heterotrimeric G proteins and through G protein-coupled receptor kinase (GRK)/β-arrestin pathways that also mediate receptor phosphorylation, desensitization, and internalization. Through a long-standing collaboration, the Stamler and Gaston groups have found that airways are regulated by nitric oxide (NO) through S-nitrosylation of thiols to form S-nitrosothiol (SNO), including on cysteine residues in proteins, a post-translational modification that alters protein functions. In addition, SNO forms on low molecular weight thiols, including glutathione to form SNO-glutathione (GSNO). We demonstrated that inhaled GSNO elevates lung protein-SNO and is protective in asthma, identified the enzyme SNO- glutathione reductase (GSNOR) that inactivates GSNO, and demonstrated that mice lacking GSNOR are protected from developing asthma. Since the β2AR can activate NO synthase in the airways to generate NO, there is a need to discover how this promotes endogenous SNO-mediated bronchoprotection. Our recent work has shown that β2AR is S-nitrosylated after activation, and preventing SNO-β2AR with a point mutation augments β2AR signaling. Importantly, mice bearing β2AR with a knock-in of this mutation are protected from developing asthma. We have previously shown that β2AR regulators GRK2 and β-arrestin2 are S-nitrosylated to inhibit their activity to desensitize the β2AR, and mice bearing GRK2-C340S and β-arrestin2-C253S knock-in exhibit heighted β2AR activity and worsened injury in cardiac models. We have begun to test the efficacy of inhaled GSNO to affect bronchorelaxation and improved lung function in patients, and these data and clinical samples uniquely position us to examine the role of S-nitrosylation in regulating the β2AR pathway from bench to bedside. The Central Hypothesis of Project 1 is that the β2AR signaling system is a key target and mediator of SNO-GSNO- GSNOR protective effects in the airways in severe asthma. Our studies will define the role of S-nitrosylation of specific β2AR signaling pathway components in a murine model of asthma, delineate the roles of inhaled GSNO and of GSNO dinitrosylases on β2AR signaling components in murine models of asthma and in human lung primary cells, and demonstrate that inhaled GSNO improves both airway flow and β2-agonist responsiveness in severe asthma. This work complements the clinical aims in Projects 2 and 3 by providing a mechanistic link between NO/GSNO/GSNOR actions and the regulation of β2AR signaling in severe asthma.

项目摘要/摘要项目1 哮喘遭受了超过7％的美国人的痛苦，这是导致气道的慢性炎症的结果重塑和反应性会阻碍空气流动。严重的哮喘是哮喘，仍然有问题尽管对常规哮喘疗法进行了最大的干预，但它占死亡率的大部分和哮喘的成本。气道β2-肾上腺素受体（β2AR）信号的功能障碍有助于严重哮喘发病机理，但是负责的确切信号传导机制尚不清楚。同时使用β2AR激活吸入的“β-激动剂”药物是急性哮喘治疗的中流，β2AR的过度激活是有害的，可以致命。气道β2AR通过异三聚体G蛋白和G蛋白偶联受体发出信号激酶（GRK）/β-arrest蛋白途径也介导受体磷酸化，脱敏和内部化。通过长期的合作，Stamler和Gaston团体发现Airways 由一氧化氮（NO）通过硫醇的S-硝基化来调节，形成S-硝基硫醇（SNO），包括ON 半胱氨酸保留在蛋白质中，这是一种改变蛋白质功能的翻译后修饰。另外，SNO 在低分子量硫醇（包括谷胱甘肽）上形成以形成Sno-Glutathione（GSNO）。我们证明了遗传GSNO升高了肺蛋白-SNO并在哮喘中受到保护，确定了SNO- 使GSNO失活的谷胱甘肽还原（GSNOR），并证明缺乏GSNOR的小鼠是免受哮喘的影响。由于β2AR可以在气道中激活NO合酶以生成NO，所以有必要发现这如何促进内源性SNO介导的支气管保护。我们最近的工作已经表明，激活后β2AR被S-亚硝基化，并防止SNO-β2AR具有点突变 β2AR信号传导。重要的是，携带β2AR的小鼠被敲入该突变受到保护哮喘。我们先前已经表明，β2AR调节剂GRK2和β-arrestin2被S-硝基化以抑制其使β2AR脱敏的活性，以及带有GRK2-C340和β-arrestin2-C253S的小鼠敲击高度 β2AR活性和心脏模型中的损伤恶化。我们已经开始测试继承GSNO的效率影响患者的支气管扫描和改善的肺功能，这些数据和临床样品独特定位我们要检查S-亚硝基化在调节从长凳到床边的β2AR途径中的作用。这项目1的中心假设是β2AR信号系统是SNO-GSNO-的关键目标和中介者严重哮喘的GSNOR保护效应。我们的研究将定义S-亚硝基化的作用特定的β2AR信号通路途径成分在哮喘的鼠模型中，描述了遗传GSNO的作用在哮喘和人肺中鼠模型中的β2AR信号成分上的GSNO二硝基酶主细胞，并证明遗传GSNO可改善气道流和β2激动剂的反应性严重的哮喘。这项工作通过提供机械链接来完成项目2和3的临床目标在NO/GSNO/GSNOR作用与严重哮喘中β2AR信号的调节之间。