Novel Regulation of Renal Function by S-Nitrosylation

S-亚硝基化对肾功能的新调节

• 批准号: 9792377
• 负责人: JONATHAN S. STAMLER
• 金额: $ 43.16万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-25 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9792377
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Animal Model; Antioxidants; Arginine; Biological Models; Cell model; Coenzyme A; Complement; Computer Simulation; Crystallization; Data; Disease; Drug Targeting; Endothelium; Enzymes; Equilibrium; Family; Glycolysis; Grant; Injury; Ischemia; Isoenzymes; Kidney; Knock-out; Knockout Mice; Mammals; Mediating; Metabolic; Metabolism; Modeling; Modification; Molecular; Mus; NADP; NOS3 gene; Nitric Oxide; Nitric Oxide Synthase; Operating System; Oxidoreductase; Pharmaceutical Preparations; Physiological; Post-Translational Protein Processing; Protein S; Proteins; Proximal Kidney Tubules; Pyruvate Kinase; Refractory; Regulation; Renal function; Renal tubule structure; Reperfusion Injury; Reperfusion Therapy; Role; S-Nitrosothiols; SKIL gene; Sepsis; Signal Transduction; Sterols; Structure; Sulfhydryl Compounds; System; Therapeutic; Tubular formation; Yeasts; base; design; drug candidate; inhibitor/antagonist; insight; member; metabolic profile; nanomolar; nephrotoxicity; novel; novel drug class; novel therapeutics; regenerative; renal ischemia; small molecule inhibitor; small molecule therapeutics; urinary tract obstruction

PROJECT SUMMARY/ABSTRACT Nitric oxide (NO), generated in the kidney by renal tubular NO synthase (eNOS), can exert renoprotective effects. However, the mechanisms by which NO does so remain poorly understood. Nitric oxide bioactivity is principally conveyed by S-nitrosylation of proteins, the oxidative modification of protein thiols by NO to form S-nitrosothiols (SNOs). Protein S-nitrosylation is reversibly regulated by enzymatic mechanisms including S-nitrosylases and denitrosylases and accumulating evidence implicates dysregulated S- nitrosylation in disease. Here, we identify a novel enzymatic machinery subserving protein S-nitrosylation in the kidney, including a novel metabolic intermediate S-nitroso-coenzyme A (SNO-CoA) that conveys NO bioactivity and its cognate reductase, SNO-CoA reductase (SCoAR), an enzyme of previously unknown function that is highly expressed in kidney proximal tubules. SCoAR mediates the breakdown of SNO-CoA thereby lowering steady-state levels of SNO-proteins (in equilibrium with SNO-CoA). Knockout of SCoAR in mice (SCoAR-/-) is shown to increase S-nitrosylation within the kidney and to protect against ischemia/reperfusion (I/R) injury, whereas deletion of eNOS (SCoAR-/-/eNOS-/-) abrogates this protection. Thus classic renoprotection by eNOS is identified with the SNO-CoA/SCoAR system. Using metabolic profiling and mass spec (MS)-based SNO-protein identification, we have found that renoprotection by the SNO-CoA/SCoAR system is likely mediated by metabolic reprogramming within the kidney. Mechanistically, our data suggest that pyruvate kinase (PKM2) is a major locus of regulation by the SNO-CoA/SCoAR system during acute kidney injury (AKI). Our grant thus explores the idea that S-nitrosylation of PKM2 is renoprotective through metabolic reprogramming that entails SCoAR-regulated coordination of fuel utilization and antioxidant/regenerative defenses. To lay the groundwork for new therapies, we have also screened for SCoAR inhibitors and successfully developed a new class of drug candidate with nanomolar potency, which will be studied herein. Thus, our discovery not only reveals the first physiological function of the SNO-CoA/SCoAR system in mammals, but promises to open a new chapter in our understanding of AKI with immediate therapeutic implications. !

项目摘要/摘要 一氧化氮（NO）是由肾小管型NO合成酶（eNOS）在肾脏中产生的， 方面的影响.然而，NO这样做的机制仍然知之甚少。一氧化氮生物活性 主要通过蛋白质的S-亚硝基化、蛋白质巯基被NO氧化修饰， 形成S-亚硝基硫醇（SNOs）。蛋白质S-亚硝基化受酶机制可逆调节 包括S-亚硝基化酶和脱亚硝基化酶，越来越多的证据表明S- 疾病中的亚硝基化在这里，我们确定了一种新的酶机制subserving蛋白质S-亚硝基化， 肾脏，包括一种新的代谢中间体S-亚硝基辅酶A（SNO-CoA）， 生物活性及其同源还原酶，SNO-CoA还原酶（SCoAR），一种以前未知的酶 在肾脏近端小管中高度表达的功能。SCoAR介导SNO-CoA的分解 从而降低SNO-蛋白质的稳态水平（与SNO-CoA平衡）。中SCoAR的敲除 小鼠（SCoAR-/-）显示增加肾脏内的S-亚硝基化，并保护免受 缺血/再灌注（I/R）损伤，而eNOS（SCoAR-/-/eNOS-/-）的缺失消除了这种保护作用。 因此，eNOS的经典肾保护作用与SNO-CoA/SCoAR系统有关。使用代谢 通过分析和基于质谱（MS）的SNO蛋白鉴定，我们发现， SNO-CoA/SCoAR系统可能是通过肾脏内的代谢重编程介导的。机械地说， 我们的数据表明丙酮酸激酶（PKM 2）是SNO-CoA/SCoAR调控的主要位点 急性肾损伤（阿基）。因此，我们的资助探索了PKM 2的S-亚硝基化是 通过代谢重编程进行肾保护，需要SCoAR调节的燃料协调 利用和抗氧化/再生防御。为了给新疗法打下基础，我们还 筛选SCoAR抑制剂，并成功开发了一类新的候选药物， 功效，将在此进行研究。因此，我们的发现不仅揭示了第一个生理功能， 哺乳动物的SNO-CoA/SCoAR系统，但有望为我们理解阿基打开新的篇章 具有直接的治疗意义。 !