Nicotinamide nucleotide transhydrogenase regulates redox balance in atherosclerosis

烟酰胺核苷酸转氢酶调节动脉粥样硬化的氧化还原平衡

基本信息

批准号：
10298827
负责人：
DAVID M KRZYWANSKI
金额：
$ 36.5万
依托单位：
LOUISIANA STATE UNIV HSC SHREVEPORT
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10298827
关键词：
Adhesions Adipose tissue Animals Antioxidants Arterial Fatty Streak Atherosclerosis Bioenergetics Biological Availability Blood Vessels CRISPR/Cas technology Cell Adhesion Molecules Cell Respiration Cell physiology Cells Clinical Clinical Trials Coronary Arteriosclerosis Data Development Diabetes Mellitus Disease Endothelial Cells Endothelium Enzymes Equilibrium Event Functional disorder Genes Glutathione Reductase Heart failure High Fat Diet Human Hydrogen Peroxide Hypertension Impairment Inflammation Inflammatory Knockout Mice Laboratories Ligation Lipids Lipoproteins Metabolic Pathway Metabolism Mitochondria Modeling Modification Morbidity - disease rate Mus NAD(P)+ transhydrogenase NADH NADP NADPH Oxidase Obesity Oxidation-Reduction Patients Phenotype Play Production Publishing Reactive Oxygen Species Regulation Role Serum System Testing United States Vascular Cell Adhesion Molecule-1 Vascular Diseases Vascular Endothelial Cell Viral antioxidant enzyme antioxidant therapy aortic arch catalase chronic inflammatory disease endothelial dysfunction genome wide association study glutathione peroxidase in vivo intercellular cell adhesion molecule knock-down mortality new therapeutic target novel obese patients overexpression oxidant stress oxidized low density lipoprotein preservation prevent recruit response small hairpin RNA success therapeutically effective trafficking vascular inflammation

项目摘要

PROJECT ABSTRACT Atherosclerosis, a progressive chronic inflammatory disease of the vessel wall, is regulated by oxidant stress throughout the course of disease development. Endothelial dysfunction is a critical, initiating step in the development of atherosclerosis and increasing evidence implicates mitochondrial reactive oxygen species (ROS) as an important contributor to endothelial dysfunction and vascular inflammation. Nicotinamide nucleotide transhydrogenase (NNT) is emerging as an important enzyme in the regulation of mitochondrial NADPH levels which can have a significant impact on a number of metabolic pathways through the regulation of mitochondrial redox balance. Preliminary data from our laboratory supports this concept demonstrating that NNT expression is reduced in the endothelium of atherosclerotic patients when compared to healthy controls. These data are consistent with previously published data from our laboratory indicating that AAV8-PCSK9 and high fat diet treatment led to increased plaque burden in C57Bl/6J mice that lack NNT. Recent studies utilizing small hairpin RNA knockdown of NNT in human aortic endothelial demonstrates increased hydrogen peroxide production that is associated with reductions in mitochondrial NADPH and impairment of downstream antioxidant enzymes glutathione peroxidase and glutathione reductase. Furthermore, the loss of NNT in these cells also led to increased adhesion molecule expression and inflammatory cell trafficking suggesting that NNT’s role in regulating mitochondrial balance can play a critical role in modulating atherosclerotic plaque development. We have confirmed these observations in vivo using a novel endothelial specific knockout mouse where the loss of NNT promotes enhanced adhesion molecule expression in the aortic arch of mice subjected to AAV8-PCSK9, partial carotid ligation, and high fat diet in a model of disturbed flow induced plaque development. Building upon these findings, we propose that the loss of NNT activity contributes to a pro-oxidative mitochondrial phenotype that exacerbates the progression of atheroscelrosis by enhancing mitochondrial ROS production, endothelial dysfunction, and vascular inflammation . To test this hypothesis, studies are proposed that will determine i) if NNT inhibits mitochondrial ROS production and preserves mitochondrial antioxidant activity and oxidative metabolisim in human vascular endothelial cells; ii) if reduced NNT expression and increased mitochondrial ROS production stimulates Nox activity that contributes to reduced NO bioavailibilty and promotes endothelial activation; and iii) if endothelial NNT critically regulates mitochondrial redox balance and vascular function in mice treated with high fat diet. Data from the proposed studies will identify NNT as a master regulator of mitochondrial function and ROS production whose absence exacerbates the development of atherosclerosis by promoting endothelial dysfunction and vascular inflammation, leading to increased plaque development.

项目摘要动脉粥样硬化是一种进行性血管壁慢性炎症性疾病，受氧化应激调节在整个疾病发展过程中，内皮功能障碍是关键的起始步骤。动脉粥样硬化的发展和越来越多的证据表明线粒体活性氧（ROS）是内皮功能障碍和血管炎症的重要贡献者。转氢酶 (NNT) 正在成为调节线粒体 NADPH 水平的重要酶它可以通过线粒体的调节对许多代谢途径产生重大影响我们实验室的初步数据支持这一概念，表明 NNT 表达。与健康对照相比，动脉粥样硬化患者的内皮细胞减少。与我们实验室之前发表的数据一致，表明 AAV8-PCSK9 和高脂肪饮食最近的研究利用小发夹，治疗导致缺乏 NNT 的 C57Bl/6J 小鼠斑块负担增加。人主动脉内皮细胞中 NNT 的 RNA 下调表明过氧化氢敲击产生增加，与线粒体 NADPH 减少和下游抗氧化酶受损有关谷胱甘肽过氧化物酶和谷胱甘肽还原酶此外，这些细胞中NNT的损失也导致。粘附分子表达和炎症细胞运输增加表明 NNT 在调节线粒体平衡可以在调节动脉粥样硬化斑块的发展中发挥关键作用。已经使用新型内皮特异性敲除小鼠在体内证实了这些观察结果，其中 NNT 促进 AAV8-PCSK9 小鼠主动脉弓中粘附分子的表达增强，部分颈动脉结扎和高脂肪饮食在血流紊乱诱导斑块形成的模型中的应用。这些发现表明，NNT 活性的丧失有助于线粒体的促氧化。通过增强线粒体 ROS 加剧动脉粥样硬化进展的表型为了验证这一假设，正在进行研究。提议将确定 i) NNT 是否抑制线粒体 ROS 产生并保护线粒体人血管内皮细胞的抗氧化活性和氧化代谢；ii) 如果 NNT 表达减少；线粒体 ROS 产生的增加会刺激 Nox 活性，从而降低 NO 生物利用度并促进内皮激活；以及 iii) 如果内皮 NNT 关键性调节线粒体氧化还原平衡拟议研究的数据将确定 NNT 是一种高脂肪饮食的小鼠的血管功能。线粒体功能和 ROS 产生的主要调节因子，其缺失会恶化通过促进内皮功能障碍和血管炎症导致动脉粥样硬化，导致斑块增加发展。