Sub-cellular Targeting of Endothelial ROS in Myocardial Ischemia

心肌缺血中内皮活性氧的亚细胞靶向

• 批准号: 10705336
• 负责人: Ruhul Abid
• 金额: $ 61.32万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-10 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10705336
• 关键词: 3-nitrotyrosine; AMP-activated protein kinase kinase; Address; Anabolism; Animal Model; Animals; Antioxidants; Arteries; Biogenesis; Biological Assay; Blood Vessels; Brain Hypoxia-Ischemia; Cardiac Surgery procedures; Cardiovascular Diseases; Cell Proliferation; Cell Survival; Chronic; Coagulation Process; Complex; Congresses; Coronary; Coronary Arteriosclerosis; Coronary Vessels; Data; Down-Regulation; Electron Transport; Electrons; Endothelial Cells; Endothelium; Energy Supply; Exposure to; Extravasation; Family suidae; Fibrin; Funding; Generations; Glucose; Glycolysis; Grant; Heart; Heart Atrium; Human; Hypoxia; Incubated; Infarction; Ischemia; Lateral; Ligation; Manuscripts; Measures; Mediating; Membrane Potentials; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Mus; Myocardial Infarction; Myocardial Ischemia; NADPH Oxidase; Nutrient; Operative Surgical Procedures; Outcome; Oxidants; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Oxygen Consumption; Patients; Production; Proteomics; Publishing; Reactive Oxygen Species; Recovery; Recovery of Function; Reporting; Research; SOD2 gene; Source; Stroke; System; Therapeutic; Tissues; Transgenes; Transgenic Animals; Transgenic Organisms; United States National Institutes of Health; Vascular Endothelium; Work; ameroid; angiogenesis; base; cardiac angiogenesis; cardiovascular disorder prevention; density; extracellular; fatty acid oxidation; genetic manipulation; heart function; improved; in vivo; in vivo evaluation; innovation; insight; mimetics; mitochondrial membrane; nanodrug; nanoparticle; nanoparticle drug; neoplastic cell; normoxia; novel; novel strategies; overexpression; phosphoproteomics; preservation; prevent; recruit; resilience; subcellular targeting; trend

renewal application is based on the novel findings that reduction in mito-ROS, using genetic manipulation and/or nanoparticle drugs, improve survival and proliferation of coronary EC and help recover cardiac function in a post-myocardial infarct (MI) animal model and human atrial tissue. Preliminary findings in the current application demonstrate that reduction of mito-ROS using transgene (MnSOD) and nanodrugs (JP4-039, XJB5-131) induce mitochondrial complex I biogenesis (proteomic and phosphoproteomic data) and oxidative phosphorylation (Ox-Phos) in EC resulting in coronary angiogenesis cardiac function recovery in post-MI heart. This innovative EC, like most tumor cells, utilizes anerobic glycolysis as a major (85%) source of ATP Thus, the shift from less- efficient energy production system glycolysis (2 ATP/glucose molecule), to a more efficient Ox-Phos (34-36 ATP/glucose) may provide critical energy support to EC needed during ischemia (low glucose, oxygen). Here, we propose to examine a novel overarching hypothesis that modulation of mito-ROS improves production, while EC mitochondria are mostly involved in dNTP biosynthesis. Δψm and ‘super-complex’ (SC) formation resulting in efficient electron transport chain (ETC) and Ox-Phos-mediated ATP generation in EC during ischemia. This shift from glycolysis to a more efficient mitochondrial Ox-Phos may provide resilience/survival to coronary EC during ‘energy crisis’ in myocardial ischemia. The therapeutic benefit of intervening on subcellular ROS level will be best realized by specific down regulation of mito-ROS in ECs that have been exposed to ischemia/hypoxia. This hypothesis will be fully tested in vivo using our novel EC-specific transgenic MnSOD (SOD-OE; mitochondrial antioxidant) animals and supported using mitochondria-specific nitroxide and nanoparticle antioxidant in large animal model (swine) and in coronary vessels from CVD patients undergoing cardiac surgery. We propose three Specific Aims. Aim 1: Elucidate the molecular mechanisms by which modulation of mitochondrial ROS protect coronary EC from oxidative stress and induce coronary angiogenesis during myocardial ischemia. Using SOD-VE transgenic animals and myocardial infarct-mimicking (LAD ligation) surgeries, Δψm, super-complex (SC) formation, oxygen consumption rate (OCR) versus extracellular acidification rate (ECAR), ATP synthesis, will be assessed. Aim 2: Determine the effects mitochondria-targeted MnSOD-mimetic (JP4-039, XJB-5-131) nanoparticles on post-infarct vessel density and recovery of cardiac function in mice and on chronic myocardial ischemia in large animals (swine). Aim 3: Elucidate the molecular mechanisms by which mitochondrial antioxidant nanoparticles (JP4-039/XJB-5-131) improve angiogenic potential of human coronary vessels (from cardiac surgery patients) ex vivo. This study using unique animal models, human atrial tissue, and mitochondria- specific antioxidant will provide novel insights into the mechanisms by which mito-ROS can improve EC survival and angiogenesis by recruiting mitochondria as an efficient energy generator during ischemia.

更新申请是基于新的发现，减少线粒体活性氧，使用遗传 操作和/或纳米颗粒药物，改善冠状动脉EC的存活和增殖，并帮助恢复 心肌梗塞（MI）后动物模型和人心房组织中的心脏功能。初步调查结果 在本申请中，证明了使用转基因（MnSOD）和纳米药物减少线粒体-ROS （JP 4 -039，XJB 5 -131）诱导线粒体复合物I生物发生（蛋白质组学和磷酸蛋白质组学数据） 和氧化磷酸化（Ox-Phos）导致冠状动脉血管生成，心功能恢复 心肌梗死后心脏 这一创新 EC，像大多数肿瘤细胞一样，利用无氧糖酵解作为ATP的主要（85%）来源 所以说，少一点的转变， 有效的能量生产系统糖酵解（2 ATP/葡萄糖分子），以更有效的Ox-Phos（34-36 ATP/葡萄糖）可以为缺血（低葡萄糖、氧）期间所需的EC提供关键的能量支持。在这里， 我们提出了一个新的总体假设，即线粒体活性氧的调节可以改善 而EC线粒体主要参与dNTP的生物合成。 Δ Δ m和 “超复合物”（SC）形成，导致有效的电子传递链（ETC）和Ox-Phos介导的 缺血时EC中ATP的产生。这种转变，从 糖酵解为更有效的线粒体Ox-Phos 可能在心肌缺血的“能量危机”期间为冠状动脉EC提供弹性/存活。 治疗 干预亚细胞ROS水平的益处最好通过特异性下调线粒体-ROS来实现 在已经暴露于缺血/缺氧的EC中。这一假设将在体内进行充分测试，使用我们的 新的EC特异性转基因MnSOD（SOD-OE;线粒体抗氧化剂）动物和使用 在大型动物模型（猪）和冠状动脉中的动脉特异性氮氧化物和纳米颗粒抗氧化剂 接受心脏手术的CVD患者的血管。我们提出三个具体目标。目标1：阐明 调节线粒体ROS保护冠状动脉EC免受氧化损伤的分子机制 在心肌缺血期间应激并诱导冠状动脉血管生成。使用SOD-VE转基因动物 和心肌梗死模拟（LAD结扎）手术，Δ m，超复合物（SC）形成，氧 将评估消耗速率（OCR）与细胞外酸化速率（ECAR）、ATP合成的关系。目的 2：确定靶向MnSOD模拟物（JP 4 -039，XJB-5-131）纳米颗粒对细胞增殖的影响。 对小鼠心肌梗死后血管密度和心功能恢复的影响，以及对慢性心肌缺血小鼠的影响。 大型动物（猪）。目的3：阐明线粒体抗氧化剂对心肌细胞凋亡的分子机制。 纳米颗粒（JP 4 -039/XJB-5-131）改善人冠状血管（来自心脏）的血管生成潜力 手术患者）离体。这项研究使用独特的动物模型，人类心房组织和线粒体- 特异性抗氧化剂将为线粒体活性氧改善EC的机制提供新的见解 存活和血管生成通过募集线粒体作为缺血期间的有效能量发生器。