Role of S-Nitrosylation on Beta-Adrenergic Signaling in Cardiac Injury and Repair

S-亚硝基化对 β-肾上腺素能信号传导在心脏损伤和修复中的作用

• 批准号: 10370376
• 负责人: Walter J. Koch
• 金额: $ 70.26万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10370376
• 关键词: ADRBK1 gene; Acute; Adrenergic Agents; Adrenergic Receptor; Affect; Aging; Arr2; Arrestins; Binding; Biological Models; Cardiac; Cardiac Myocytes; Cardiovascular system; Catecholamines; Cell physiology; Cessation of life; Chronic; Collaborations; Cyclic GMP-Dependent Protein Kinases; Cysteine; Data; Defect; Disease; Down-Regulation; Endothelium; Enzymes; Evaluation; Functional disorder; G Protein-Coupled Receptor Signaling; G protein coupled receptor kinase; G-Protein-Coupled Receptors; Goals; Guanylate Cyclase; Health; Heart; Heart Injuries; Heart failure; Hybrids; Impairment; Injury; Knock-in Mouse; Laboratories; Lead; Mediating; Modeling; Modification; Mus; Muscle Cells; Mutant Strains Mice; Myocardial; Myocardial Ischemia; Myocardial dysfunction; Myocardium; Neurons; Nitric Oxide; Nitric Oxide Synthase; Organ; Outcome; Oxidation-Reduction; Pathogenesis; Pathologic; Pathology; Phosphorylation; Phosphotransferases; Physiological; Physiology; Play; Post-Translational Protein Processing; Property; Protein Isoforms; Protein S; Proteins; Reagent; Receptor Signaling; Refractory; Regulation; Role; S-Nitrosothiols; Signal Transduction; Site; Stress; Sulfhydryl Compounds; System; Testing; Therapeutic; Therapeutic Intervention; Translations; Work; base; beta-adrenergic receptor; body system; cardiogenesis; cardioprotection; desensitization; heart function; improved; injury and repair; innovation; insight; ischemic injury; mouse model; mutant; novel; novel therapeutic intervention; receptor; receptor function; repaired; response; response to injury

SUMMARY: During the development of heart failure (HF), especially after ischemic injury, derangements in myocardial β- adrenergic receptor (βAR) signaling contribute centrally to pathogenesis, including signal uncoupling and receptor desensitization leading to myocyte death and contractility defects. Waning of signaling through cardiac βARs and other G protein-coupled receptors (GPCRs) is classically regulated via receptor phosphorylation and internalization mediated by GPCR kinases (GRKs) and β-arrestins (β-Arrs). This is significant since the activity of GRK2, which is elevated in myocardium after injury/stress, is pathologic in HF and its inhibition is therapeutic. Through a long-standing collaboration, the Koch and Stamler laboratories have found that GPCRs are regulated by nitric oxide (NO), through S-nitrosylation of cysteine to form protein S-nitrosothiol (SNO), including profound regulation of GRK2 and of β-Arr2. Since cardiac GPCRs, including all three βARs, can activate NO synthase (NOS) enzymes, there is a need to discover how this can promote SNO-mediated cardioprotection, especially downstream of β2- and β3ARs. Our prior work has shown that NO from endothelial NOS (eNOS) inhibits GRK2 by S-nitrosylation at Cys340. Loss of SNO-based regulation in GRK2-C340S mutant knock-in (KI) mice leads to un-checked and enhanced GRK2 activity, and to increased ischemic injury, and to dysfunction during aging. Our labs have also shown that neuronal/inducible NOS (n/iNOS) activity can regulate β-Arr2 through SNO-Cys253 to maintain physiological βAR signaling in the heart. The loss of this SNO-β-Arr2 regulation in β-Arr2-C253S KI mice leads to increased βAR desensitization and HF. Additionally, the Stamler lab recently discovered that β2AR is S-nitrosylated at Cys265 and that this modification regulates β2AR desensitization. Together these data suggest tightly integrated regulation of βAR/GPCR function via receptor-stimulated S-nitrosylation, which plays a central but largely unappreciated role in controlling myocardial function. Our data provides novel insight into consequences of the nitroso-redox imbalance in failing heart. The Central Hypothesis of this Multi-PI proposal is that cardiac βAR signaling and desensitization via GRK2 and β-Arr2 are regulated by S-nitrosylation and that nitroso-redox stress can be understood in terms of altered SNO of receptor, GRK and β-Arr to significantly impair the heart’s response to injury. Specific Aims are: [1] To determine whether GRK2 inhibition via S-nitrosylation plays a mechanistic role in selective βAR responses during cardiac ischemic injury; [2] To determine if β2AR is S-nitrosylated in the ischemic heart and whether this impacts injury and repair; [3] To determine if regulation of β-Arr2 by S-nitrosylation tunes βAR responses during cardiac dysfunction after injury and is integrated with β2AR and GRK2 SNO regulation. Successful completion of these studies will illuminate the role of S-nitrosylation in the integrated adrenergic response to cardiac injury and repair. These fundamental discoveries will reveal new insights into the regulation of cardiac function in health and disease by defining therapeutic interventions to promote cardioprotection, and to serve as a paradigm for signaling systems in other organs and diseases.

概括： 在心力衰竭（HF）的发展过程中，尤其是在缺血性损伤之后，心肌β-的进化 肾上腺素受体（βAR）信号转导促进发病机理，包括信号解偶联和 受体脱敏导致肌细胞的死亡和收缩性缺陷。通过心脏的信号降低 βAR和其他G蛋白偶联受体（GPCR）通过受体磷酸化和 由GPCR激酶（GRK）和β-arrestin（β-arrs）介导的内在化。这很重要，因为活动 损伤/应激后心肌中升高的GRK2的of病理学在HF中是病理性的，其抑制作用是治疗性的。 通过长期的合作，科赫和斯坦勒实验室发现GPCR受到监管 通过一氧化氮（NO），通过半胱氨酸的S-硝基化形成蛋白质S-硝基硫醇（SNO），包括深刻 GRK2和β-arr2的调节。由于心脏GPCR（包括所有三个βAR）可以激活NO合酶 （nos）酶，有必要发现这如何促进SNO介导的心脏保护，尤其是 β2-和β3AR的下游。我们先前的工作表明，内皮NOS（ENOS）的NO抑制GRK2 通过Cys340的S-硝基化。在GRK2-C340S突变敲入（Ki）小鼠中基于SNO的调节损失导致 未检查和增强的GRK2活性，增加缺血性损伤，并在衰老期间功能障碍。我们的 实验室还表明，神经元/诱导的NOS（N/INOS）活性可以通过SNO-CYS253调节β-arr2 保持心脏中的物理β信号传导。 β-arr2-C253S ki中这种SNO-β-arr2调节的丢失 小鼠导致βAR脱敏和HF增加。此外，Stamler Lab最近发现β2AR 在Cys265处被S-亚硝基化，并且这种修饰调节β2AR脱敏。这些数据在一起 建议通过受体刺激的S-硝基化对βAR/GPCR功能的紧密整合调节，该硝基化的作用 在控制心肌功能中的中心但很大程度上没有批准的作用。我们的数据提供了新的见解 硝基 - 雷克斯不平衡心脏失败的后果。该多PI提案的中心假设 是通过S-亚硝基化调节了通过GRK2和β-Arr2通过GRK2和β-Arr2脱敏的心脏β信号传导和脱敏的 可以通过改变受体，grk和β-arr的SNO明显损害的硝基 - 雷斯胁迫来理解 心脏对伤害的反应。具体目的是：[1]确定是否通过S-硝基化抑制GRK2是否 在心脏缺血性损伤期间选择性βAR反应中起着机理作用。 [2]确定β2AR是否为 缺血性心脏中的S-亚硝基化，这是否会影响损伤和修复； [3]确定是否调节 损伤后心功能障碍期间S-亚硝基化调音βAR反应的β-arr2β-arr2，并与β2AR整合 和GRK2 SNO调节。这些研究的成功完成将阐明S-硝基化在 对心脏损伤和修复的综合肾上腺素反应。这些基本发现将揭示新的 通过将治疗干预措施定义为健康和疾病中心脏功能的调节 促进心脏保护，并充当其他器官和疾病中信号系统的范式。