HEMODYNAMICS AND MOLECULAR GENETIC RISK FACTORS IN ATHEROSCLEROSIS

动脉粥样硬化的血流动力学和分子遗传风险因素

• 批准号: 7056675
• 负责人: MICHAEL A GIMBRONE
• 金额: $ 39.68万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7056675
• 关键词: antioxidants; apolipoprotein E; atherosclerosis; atherosclerotic plaque; biomechanics; cardiovascular disorder risk; gene expression; genetic susceptibility; hemodynamics; high throughput technology; laboratory mouse; serial analysis of gene expression; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): The early lesions of atherosclerosis, in humans subjects and experimental animals, consistently develop in a non-random pattern in the arterial vasculature, and the geometry of these "lesion-prone areas" correlates with branch points, curvatures and other regions of altered blood flow. Previous studies in our laboratory have demonstrated direct effects of biomechanical forces on the expression of pathophysiologically relevant genes by the endothelial cell (EC). This has led us to hypothesize that hemodynamic forces, in particular wall shear stresses generated by steady laminar and disturbed laminar flow, can function as both positive and negative stimuli in atherogenesis, via effects on EC gene expression. To test this hypothesis, we propose a series of interrelated in vitro and in vivo experiments, utilizing a combination of cell biological, molecular biological and experimental pathological techniques under three Specific Aims. In the First Specific Aim, we will extend and critically test this hypothesis (established in principle during the previous project period) by utilizing a newly developed "arterial waveform" flow device to simulate the fluid mechanics of pulsatile human blood flow on cultured human endothelial cells in vitro, and define patterns of gene expression via high-throughput transcriptional profiling techniques and analytical methods previously developed in this project. In the Second Specific Aim, we will validate in vivo, patterns of expression of putative pathophysiologically relevant endothelial genes, identified via sentinel cluster analysis of in vitro transcriptional profiling data in Specific Aim 1, directly in "high-probability" and "low-probability" regions of the mouse aorta, in atherosclerosis-prone strains, and in appropriate normal and diseased human arterial tissue samples. In the Third Specific Aim, we will critically test the hypothesis that shear-regulated endothelial gene expression contributes to athero-susceptibility, using various transgenic strategies to modulate expression of candidate pathogenic and/or protective shear-regulated genes in murine models of atherosclerosis, and observe the effects on lesion localization and progression/regression. The proposed studies thus focus on a fundamental aspect of the pathobiology of atherosclerosis - the role of biomechanical forces as a determinant of the geometric pattern of lesion formation. The combination of high-throughput genomics with sentinel-cluster analysis, and in vitro modeling with in vivo validation, promises to provide new insights into disease mechanisms.

描述（由申请人提供）： 动脉粥样硬化的早期病变，在人类受试者和实验 动物在动脉脉管系统中以非随机模式持续发展，并且这些"病变倾向区域"的几何形状与分支点、曲率和 其他区域的血流改变。我们实验室以前的研究 证明了生物力学力对表达的直接影响， 内皮细胞（EC）的病理生理相关基因。这 使我们假设血液动力学力，特别是壁剪切力 稳定层流和扰动层流产生的应力， 作为动脉粥样硬化形成的正性和负性刺激，通过对EC基因的影响， 表情为了验证这一假设，我们提出了一系列相互关联的假设。 体外和体内实验，利用细胞生物学， 分子生物学和实验病理学技术 具体目标。在第一个具体目标中，我们将扩展并严格测试 这一假设（原则上在上一个项目期间确立） 通过利用新开发的"动脉波形"流动装置来模拟 人血管内皮细胞上脉动血流的流体力学研究 体外细胞，并通过高通量确定基因表达模式 转录谱分析技术和分析方法， 在这个项目中开发的。在第二个具体目标中，我们将验证 体内，假定的病理生理学相关的表达模式 内皮细胞基因，通过体外的前哨聚类分析确定 转录谱数据在特定目标1，直接在"高概率" 和小鼠主动脉的"低概率"区域， 动脉粥样硬化倾向菌株，并在适当的正常和患病的人 动脉组织样本在第三个具体目标中，我们将严格检验 剪切调节的内皮基因表达有助于 动脉粥样硬化易感性，使用各种转基因策略来调节 候选的致病性和/或保护性剪切调节基因的表达 建立小鼠动脉粥样硬化模型，观察其对动脉粥样硬化病变的影响 定位和进展/回归。因此，拟议的研究重点是 动脉粥样硬化病理生物学的基本方面- 生物力学力作为病变几何模式的决定因素 阵高通量基因组学与哨兵簇的结合 分析，并在体外建模与体内验证，承诺提供 对疾病机制的新见解。