GAPDH, DNA Repair and Atherosclerosis

GAPDH、DNA 修复和动脉粥样硬化

• 批准号: 10210430
• 负责人: Sergiy Sukhanov
• 金额: $ 38万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10210430
• 关键词: Acute; Apoptosis; Apoptotic; Arterial Fatty Streak; Arteries; Atherosclerosis; Binding; Blood Vessels; Cardiovascular Diseases; Carotid Artery Plaques; Cells; Collagen; Complementary DNA; Coronary heart disease; DNA; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA Repair Pathway; Data; Dependovirus; Development; Diagnostic; Disease; Down-Regulation; Enzymes; Event; Genomic Instability; Glyceraldehyde-3-Phosphate Dehydrogenases; Glycolysis; Goals; Heart; Homeostasis; Human; Lesion; Link; Lipids; Mediating; Molecular; Morbidity - disease rate; Mus; Necrosis; Neurodegenerative Disorders; Nuclear; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxides; Peptides; Pharmaceutical Preparations; Play; Prevention; Role; Rupture; Signal Transduction; Smooth Muscle Myocytes; Specimen; Testing; Therapeutic; Thick; Thinness; Vascular Smooth Muscle; atherogenesis; atheroprotective; base; endonuclease; gain of function; healthy lifestyle; improved; in vivo; in vivo evaluation; innovation; loss of function; mortality; mouse model; new therapeutic target; novel; overexpression; oxidized low density lipoprotein; prevent; repaired; sensor; vector

Project Summary/Abstract Accumulated DNA damage is recognized as a causal factor in the initiation and progression of atherosclerosis. Genomic instability in the vascular smooth muscle cells (SMC) leads to cell apoptosis and contributes to atherosclerotic plaque vulnerability. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is the major cellular sensor ultimately responsible for maintaining of cellular homeostasis, however its specific role in atherogenesis is completely unknown. It has been shown that a key pro-atherogenic lipid OxLDL downregulated GAPDH in human aortic SMC and that GAPDH expression was markedly decreased in the atherosclerotic plaque SMC and that low GAPDH level was associated with increased apoptosis. GAPDH reduced DNA damage and suppressed SMC apoptosis via a novel molecular mechanism involving nuclear GAPDH interaction with apurinic/apyrimidinic endonuclease 1 (Ape1), the major oxidized DNA repair enzyme. SMC-specific GAPDH overexpression decreased DNA damage, reduced plaque SMC apoptosis and decreased atherosclerotic burden. Importantly, atherosclerotic plaques in GAPDH-overexpressing mice had elevated SMC levels, increased collagen, reduced necrotic cores and thicker SMC-rich fibrous caps suggesting enhanced plaque stability. The major focus of the current proposal is to study the mechanism mediating GAPDH-induced anti-atherosclerotic and plaque stabilizing effect and to determine whether GAPDH mimicking peptide (GMP) reduces atherosclerotic burden and improve plaque stability in atherosclerotic mice. The main hypothesis is that GAPDH reduces atherosclerotic burden and enhances features of plaque stability via stimulation of Ape1-dependent DNA repair and suppression of SMC apoptosis. This will test the hypothesis in following Specific Aims: Specific Aim 1: To demonstrate that SMC-specific GAPDH regulates DNA repair, apoptosis, atherosclerotic burden and features of plaque stability and identify mechanism. Specific Aim 2: To determine whether GAPDH-mimicking peptide (GMP) will activate Ape1, stimulate DNA repair, suppress cell apoptosis and reduce atherosclerosis. Proposal will use adeno-associated viruses (AAVs) to perform SMC-targeted GMP cDNA transfer in atherosclerotic mice. AAV-based vectors are approved to use in humans, therefore the long-term goal is to use the AAV-GMP vector as an innovative pro-DNA repair and anti-apoptotic therapy to treat unstable atherosclerosis. Studying of anti-atherosclerotic effects of GAPDH and demonstration that GMP induces plaque-stabilizing effect will lead to the development of novel targeted therapies to treat atherosclerosis and prevent acute vascular events.

项目总结/摘要 累积的DNA损伤被认为是动脉粥样硬化发生和发展的一个致病因素。 血管平滑肌细胞（SMC）中基因组的不稳定性导致细胞凋亡， 动脉粥样硬化斑块的脆弱性。甘油醛3-磷酸脱氢酶（GAPDH）是主要的 细胞传感器最终负责维持细胞内稳态，然而其在 动脉粥样硬化形成是完全未知的。已经表明，一种关键的促动脉粥样硬化脂质OxLDL 在人主动脉平滑肌细胞中GAPDH表达下调， 动脉粥样硬化斑块SMC和GAPDH水平降低与细胞凋亡增加相关。GAPDH 通过一种新的分子机制减少DNA损伤和抑制SMC凋亡， GAPDH与脱嘌呤/脱嘧啶核酸内切酶1（Ape 1）的相互作用，Ape 1是主要的氧化DNA修复酶。 SMC特异性GAPDH过表达减少DNA损伤，减少斑块SMC凋亡， 降低动脉粥样硬化负担。重要的是，GAPDH过表达小鼠的动脉粥样硬化斑块 SMC水平升高，胶原蛋白增加，坏死核心减少，富含SMC的纤维帽变厚，表明 增强斑块稳定性。目前建议的主要重点是研究调解机制 GAPDH诱导的抗动脉粥样硬化和斑块稳定作用，并确定是否GAPDH模拟 在动脉粥样硬化小鼠中，GMP肽降低动脉粥样硬化负荷并改善斑块稳定性。主要 假设GAPDH通过以下途径降低动脉粥样硬化负荷并增强斑块稳定性特征： 刺激Ape 1依赖的DNA修复和抑制SMC凋亡。这将测试假设在 具体目标： 具体目的1：证明SMC特异性GAPDH调节DNA修复、凋亡， 动脉粥样硬化负荷和斑块稳定性的特点及识别机制。 具体目标2：为了确定GAPDH模拟肽（GMP）是否会激活Ape 1，刺激Ape 1的表达， DNA修复，抑制细胞凋亡，减轻动脉粥样硬化。 该提案将使用腺相关病毒（AAV）进行SMC靶向GMP cDNA转移， 动脉粥样硬化小鼠基于AAV的载体被批准用于人类，因此长期目标是使用 AAV-GMP载体作为一种创新性pro-DNA修复和抗凋亡疗法， 动脉粥样硬化GAPDH抗动脉粥样硬化作用的研究及GMP诱导的实验验证 斑块稳定作用将导致开发新的靶向疗法来治疗动脉粥样硬化， 预防急性血管事件。