Sub-cellular Targeting of Endothelial ROS in Myocardial Ischemia

续订申请是基于新的发现，即减少有丝分裂-ROS，使用基因 操纵和/或纳米药物，改善冠状动脉内皮细胞的存活和增殖并帮助恢复 心肌梗死(MI)后动物模型和人类心房组织的心功能。初步调查结果 在目前的应用中证明了使用转基因(MnSOD)和纳米药物减少有丝分裂-ROS (JP4-039，XJB5-131)诱导线粒体复合体I的生物发生(蛋白质组和磷蛋白质组数据) 内皮细胞氧化磷酸化(Ox-PHOS)导致冠脉血管生成和心功能恢复 在心肌梗死后的心脏。 这是一项创新 EC像大多数肿瘤细胞一样，利用厌氧糖酵解作为ATP的主要来源(85%) 因此，从较少的转变- 高效的能量产生系统糖酵解(2个ATP/葡萄糖分子)，转化为更有效的Ox-Phos(34-36 ATP/葡萄糖)可能为缺血(低糖、低氧)所需的内皮细胞提供关键的能量支持。这里, 我们建议检验一个新的总体假设，即MITO-ROS的调制可以改善 而EC线粒体主要参与dNTP的生物合成。 Δψm和 形成高效的电子传输链(ETC)的超络合物(SC)和Ox-Phos介导的 缺血时内皮细胞产生三磷酸腺苷。这一转变从 糖酵解为更有效的线粒体Ox-PHOS 在心肌缺血的“能量危机”期间，可能为冠脉内皮细胞提供韧性/存活率。 治疗性 干预亚细胞内ROS水平的好处将通过对MITO-ROS的特异性下调来实现 在已经暴露于缺血/缺氧的内皮细胞中。这一假设将在体内得到充分验证，使用我们的 新型EC特异性转基因MnSOD动物及其支持应用 线粒体特异性氮氧化物和纳米抗氧化剂在大动物模型(猪)和冠状动脉中的作用 接受心脏手术的心血管疾病患者的血管。我们提出了三个具体目标。目标1：澄清 线粒体ROS调节保护冠状动脉内皮细胞免受氧化损伤的分子机制 在心肌缺血期间应激并诱导冠脉血管生成。利用超氧化物歧化酶-维生素E转基因动物 和模拟心肌梗死(LAD结扎)手术，Δψm，超复合体(SC)形成，氧气 消耗率(OCR)与胞外酸化率(ECAR)，即ATP合成，将进行评估。目标 2：确定线粒体靶向MnSOD模拟物(JP4-039，XJB-5-131)纳米粒对 小鼠心肌梗死后血管密度与心功能恢复及慢性心肌缺血 大型动物(猪)。目的3：阐明线粒体抗氧化剂的分子机制 纳米粒(JP4-039/XJB-5-131)提高人冠状动脉血管新生能力 手术患者)体外。这项研究使用了独特的动物模型、人类心房组织和线粒体- 特定的抗氧化剂将为MITO-ROS改善EC的机制提供新的见解 通过招募线粒体作为缺血期间的有效能量生成器来存活和血管生成。

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.