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血管壁的进行性慢性炎症性疾病，受氧化应激的调节通过疾病发展。内皮功能障碍是在动脉粥样硬化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注射的重要贡献者。烟酰胺核苷酸在线粒体NADPH水平的调节中，经氢化酶（NNT）作为一种重要酶出现通过线粒体调节，这可能会对许多代谢途径产生重大影响氧化还原平衡。我们实验室的初步数据支持这个概念，表明NNT表达与健康对照相比，动脉粥样硬化患者的内皮减少了。这些数据是与我们实验室先前发布的数据一致，表明AAV8-PCSK9和高脂饮食治疗导致缺乏NNT的C57BL/6J小鼠的斑块Burnen增加。最近使用小发夹的研究人主动脉内皮示范中NNT的RNA敲低增加了过氧化氢的产生与线粒体NADPH的减少和下游抗氧化剂损伤有关谷胱甘肽过氧化物酶和谷胱甘肽减少。此外，这些细胞中NNT的丢失也导致增加的粘附分子表达和炎性细胞运输表明NNT在调节线粒体平衡可以在调节动脉粥样硬化斑块发育中起关键作用。我们已经使用一种新型的内皮特异性基因敲除小鼠在体内证实了这些观察结果 NNT促进了受AAV8-PCSK9的小鼠主动脉弓的增强的粘合分子表达，部分颈动脉连接和高脂饮食在流动障碍的模型中引起的牙菌斑发育。建立这些发现，我们建议NNT活性的丧失有助于促氧化线粒体通过增强线粒体ROS来加剧动脉镜的进展的表型产生，内皮功能障碍和血管炎症。为了检验这一假设，研究是提出的将确定i）如果NNT抑制线粒体ROS的产生并保留线粒体人血管内皮细胞中的抗氧化活性和氧化代谢； ii）如果降低了NNT表达线粒体ROS产生的增加会刺激NOX活性，导致无生物掌声减少并促进内皮激活； iii）如果内皮NNT严重调节线粒体氧化还原平衡用高脂饮食治疗的小鼠中的血管功能。来自拟议研究的数据将确定为NNT 线粒体功能和ROS产生的主要调节剂，其缺失使发展的发展加剧通过促进内皮功能障碍和血管感染，动脉粥样硬化，导致斑块增加发展。