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），通过肾小管NO合酶（ENOS）在肾脏中产生，可以发挥肾脏保护性 效果。但是，没有这样的机制仍然很熟悉。一氧化氮生物活性 主要是通过蛋白质的S-亚硝基化传达 形成S-硝基硫醇（SNOS）。蛋白质S-硝基化受酶促机制可逆调节 包括S-亚硝基酶和硝酸酶以及积累的证据表明S-的失调失调 硝基化疾病。在这里，我们确定了一种新型的酶促机械固定在 肾脏，包括一种新型代谢中间的S-硝基辅酶A（SNO-COA），它传达了NO 生物活性及其同源还原酶SNO-COA还原酶（SCOAR），这是先前未知的酶 在肾近端小管中高度表达的功能。 Scoar介导了Sno-COA的崩溃 从而降低了SNO蛋白质的稳态水平（与SNO-COA平衡）。淘汰赛 显示小鼠（SCOAR - / - ）显示会增加肾脏内的S-亚硝基化并防止 缺血/再灌注（I/R）损伤，而eNOS（SCOAR - / - / - /ENOS - / - ）的删除却消除了这种保护。 因此，通过SNO-COA/SCOAR系统鉴定出ENOS经典的肾脏保护。使用代谢 分析和质量规格（MS）基于SNO蛋白的识别，我们发现该重骨均受 SNO-COA/SCOAR系统可能是通过肾脏内代谢重编程介导的。机械上， 我们的数据表明，丙酮酸激酶（PKM2）是SNO-COA/SCOAR调节的主要轨迹 急性肾脏损伤期间的系统（AKI）。因此，我们的赠款探讨了PKM2的S-亚硝基化的想法 通过代谢重新编程进行重新保护 利用和抗氧化/再生防御。为了为新疗法奠定基础，我们也有 筛选为SCOAR抑制剂，并成功地开发了一种新的纳摩尔药物候选药物 效力，将在此研究。因此，我们的发现不仅揭示了 哺乳动物中的SNO-COA/SCOAR系统，但有望在我们对Aki的理解中打开新的篇章 具有直接的治疗意义。 呢