Mitochondrial Modulation of Endothelial Phenotype

内皮表型的线粒体调节

• 批准号: 7137141
• 负责人: John F. Keaney
• 金额: $ 38.84万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7137141
• 关键词: atherosclerosis; biological signal transduction; cell biology; cell component structure /function; cell migration; cell proliferation; free radical oxygen; gene expression; genetically modified animals; homeostasis; laboratory mouse; mitochondria; nitric oxide; oxidation reduction reaction; oxidative stress; phenotype; tissue /cell culture; vascular cell adhesion molecule; vascular endothelium

The endothelium is an important component of normal vascular homeostasis. It is known that normal endothelial function is disturbed in the setting of atherosclerosis and its risk factors such as hypercholesterolemia, diabetes, and hypertension. It is likely that multiple mechanisms contribute to impaired endothelial function, however, oxidant stress in the form of reactive oxygen species (ROS) production appear particularly important. It is also now widely appreciated that ROS act as signaling molecules that contribute to the vascular injury response, but in atherosclerosis ROS signaling becomes dysregulated and contributes to endothelial dysfunction. Despite this knowledge, the mechanisms of ROS signaling in the endothelium remain obscure. This proposal is based upon the central hypothesis that mitochondria are an important component of redox-sensitive signaling and, as a consequence, are a key determinant of endothelial cell phenotype. The objective of this application, therefore, is to determine the role of the mitochondrion in modulating endothelial cell phenotype and elucidate any operative mechanisms. To accomplish this goal, we first will undertake a detailed examination of how endothelial cell phenotype modulates mitochondrial functions such as protonmotive force (delta-muH+), mitochondrial ROS, and uncoupling protein expression. Using this knowledge and reagents we have developed, we will then manipulate specific mitochondrial functions (delta-muH+, UCPs) in cultured endothelial cells and determine the implications for endothelial functions known to involve ROS, such as nitric oxide bioactivity, cell proliferation and migration, and adhesion molecule expression. Because cells imperfectly model events in vivo, we will also manipulate mitochondrial function in vivo using UCP-2 null mice and mice we will develop with inducible endothelial cell-specific over-expression of UCP-2. We will then go on to determine the implications of UCP-2 manipulation endothelial phenotype in vivo both in the resting state and in response to stress in the form of arterial injury. Successful completion of these studies will provide mechanistic information on redox-mediated control of endothelial cell phenotype and afford us the necessary insight to design new therapeutic strategies that focus on improving vascular homeostasis in the setting of atherosclerosis and its risk factors.

内皮是正常血管稳态的重要组成部分。据了解，正常情况下 动脉粥样硬化及其危险因素（例如， 高胆固醇血症、糖尿病和高血压。多种机制可能会导致受损 然而，内皮功能会出现活性氧 (ROS) 产生形式的氧化应激 特别重要。现在人们还广泛认识到，ROS 作为信号分子，有助于 血管损伤反应，但在动脉粥样硬化中，ROS 信号变得失调并有助于 内皮功能障碍。尽管有了这些知识，内皮细胞中 ROS 信号传导的机制仍然存在 朦胧。该提议基于线粒体是重要的线粒体这一中心假设。 氧化还原敏感信号传导的组成部分，因此是内皮细胞的关键决定因素 细胞表型。因此，本应用的目的是确定线粒体在 调节内皮细胞表型并阐明任何操作机制。为了实现这一目标，我们 首先将详细检查内皮细胞表型如何调节线粒体功能 例如质子动力 (delta-muH+)、线粒体 ROS 和解偶联蛋白表达。使用这个 利用我们开发的知识和试剂，我们将操纵特定的线粒体功能（delta-muH+， UCP）在培养的内皮细胞中，并确定对已知涉及的内皮功能的影响 ROS，例如一氧化氮生物活性、细胞增殖和迁移以及粘附分子表达。 由于细胞不能完美地模拟体内事件，我们还将使用以下方法操纵体内线粒体功能 我们将开发 UCP-2 缺失小鼠和可诱导内皮细胞特异性过度表达 UCP-2 的小鼠。 然后我们将继续确定 UCP-2 操纵内皮表型在体内的影响 静息状态以及对动脉损伤形式的压力的反应。成功完成这些研究 将提供有关氧化还原介导的内皮细胞表型控制的机制信息，并为我们提供 设计新的治疗策略的必要见解，重点是改善血管内稳态 动脉粥样硬化的背景及其危险因素。