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，人类受试者和实验性病变 动物在动脉脉管系统中始终以非随机模式发展，这些“易病变区域”的几何形状与分支点，曲率和 其他改变血流的区域。我们实验室以前的研究已经 展示了生物力学对表达的直接影响 内皮细胞（EC）与病理生理相关的基因。这就是 导致我们假设血流动力学，特别是壁剪 稳定的层流和干扰层流产生的应力，可以起作用 作为动脉粥样硬化中的阳性刺激和阴性刺激，通过对EC基因的影响 表达。为了检验这一假设，我们提出了一系列相互关联的 体外和体内实验，利用细胞生物学的组合， 分子生物学和实验病理技术在三个 具体目标。在第一个特定目标中，我们将扩展并进行批判性测试 该假设（原则上是在上一个项目期间建立的） 通过利用新开发的“动脉波形”流动设备模拟 脉动性人体血流的流体力学在培养的人内皮上 体外细胞，并通过高通量定义基因表达的模式 先前的转录分析技术和分析方法 在这个项目中开发。在第二个特定目标中，我们将在 体内，假定的病理生理相关的表达模式 内皮基因，通过哨兵簇分析的体外分析鉴定 在“高概率”中直接在特定目标1中的转录分析数据 和小鼠主动脉的“低概率”区域 动脉粥样硬化易菌株，并以适当的正常和患病人类 动脉组织样品。在第三个特定目标中，我们将进行批判性测试 剪切调节的内皮基因表达的假设有助于 使用各种转基因策略调节动脉敏感性 候选病原体和/或保护性剪切基因中的表达 动脉粥样硬化的鼠模型，观察对病变的影响 本地化和进程/回归。因此，提出的研究集中于 动脉粥样硬化病理生物学的基本方面 - 生物力学力是病变几何模式的决定因素 形成。高通量基因组学与哨兵群的结合 分析和体内验证的体外建模有望提供 对疾病机制的新见解。