Microvascular endothelial cell heterogeneity in the central nervous system

中枢神经系统微血管内皮细胞异质性

• 批准号: 7342513
• 负责人: Joel S Pachter
• 金额: $ 16.19万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-22 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7342513
• 关键词: Address; Affect; Area; Attention; Behavior; Biochemistry; Biological; Blood; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Brain; Capillary Endothelial Cell; Cerebral cortex; Cerebrum; Classification; Condition; Coupling; DNA; DNA Microarray Chip; DNA Microarray format; Data; Depth; Development; Disease; Drug Formulations; Endothelial Cells; Evaluation; Exploratory/Developmental Grant; Gene Expression; Gene Expression Profiling; Genes; Goals; Grant; Health; Heterogeneity; Homeostasis; In Situ; Inflammatory; Investigation; Lead; Light; Location; Microarray Analysis; Modeling; Molecular; Molecular Profiling; Nature; Neuraxis; Neurons; Parietal Lobe; Pattern; Peripheral; Permeability; Physiological; Population; Process; RNA; Rate; Reporting; Reproducibility; Research; Risk; Sampling; Source; Spinal Cord; Staging; Support Contracts; System; Techniques; Testing; Tissues; Transcript; Trees; United States National Institutes of Health; arteriole; base; capillary; design; hemodynamics; laser capture microdissection; neurovascular unit; novel; tissue culture; tool; vascular bed; venule

The issue of microvascular endothelial heterogeneity greatly impacts vascular function in both health and disease, and potentially severly complicates the use of tissue culture-based models to study endothelial behavior. Yet, despite this topic having been explored in considerable depth in peripheral vascular beds, surprisingly little attention has yet been focused on the microvascular axis of central nervous system (CNS). Unrelated reports describing particular genes have, nonetheless, provocatively suggested that a diversity in endothelial gene expression may exist at the segmental level (i.e., between arterioles, capillaries and venules) as well as the regional level of the CNS, and approximate or surpass that found in the periphery. Given the preeminance of the microvascular network in forming both the blood-brain barrier and blood- spinal cord barrier, and the significant microvascular involvement in inflammatory, infectious, degenerative and traumatic conditions of the CNS, it is critical that there be a systematic and detailed evaluation of this cliinically important issue. It is of further importance that such analysis be performed on microvascular tissue in situ," as endothelial cell gene expression is exquisitely prone to environmental modulation. Accordingly, we propose the following Specific Aims: 1) To validate and optimize the approach of coupling laser capture microdissection (LCM) of brain microvascular endothelial cells, with global gene expression profiling byDMA microarray analysis. This will include demonstrating feasibility, by maximizing the percentage of transcripts detected (P call rate) in capillaries only, and establishing reproducibility, by identifying both technical and biological sources of variance in microarray data; and 2) To use the LCM/microarray approach to compare the global gene expression profiles of endothelial cells from capillaries, venules and arterioles. The R21 format chosen is specifically designed for projects that are exploratory and may involve considerable risk, but lead to the development of novel techniques that could have major impact on a field of research. In this regard, results obtained here will set the stage for evaluating regional gene expression by the neurovascular unit (i.e,endothelial cells and intimately associated neural cells) along the CNS microvascular tree in both health and disease. In turn, these studies will enable formulation of more precise endothelial models to study the molecular basis of physiological and pathophysiological processes of the cerebral microvasculature.

微血管内皮细胞异质性问题对健康和健康的血管功能都有很大影响。 疾病，并可能使基于组织培养的模型用于研究内皮细胞的使用严重复杂化 行为。然而，尽管在外周血管床中已经相当深入地探讨了这一主题， 令人惊讶的是，很少有人关注中枢神经系统(CNS)的微血管轴。 然而，描述特定基因的无关报告已经挑衅性地表明， 血管内皮细胞基因的表达可能存在于节段性水平(即，小动脉、毛细血管和 小静脉)以及中枢神经系统的区域水平，并接近或超过在外围发现的。 鉴于微血管网络在形成血脑屏障和血-脑屏障方面的优势， 脊髓屏障和显著的微血管参与炎症、感染性、退行性变 对于中枢神经系统的创伤状况，至关重要的是对此进行系统和详细的评估 具有重要意义的问题。更重要的是，对微血管组织进行这种分析 AS内皮细胞基因的表达非常容易受到环境调节的影响。因此， 我们提出了以下具体目标：1)验证和优化耦合激光捕获方法 脑微血管内皮细胞的显微解剖(LCM)，并用DMA分析全球基因表达谱 微阵列分析。这将包括通过最大限度地提高成绩单的百分比来展示可行性 仅在毛细血管中检测到(P呼叫率)，并通过确定技术和 微阵列数据的生物变异来源；以及2)使用LCM/微阵列方法进行比较 毛细血管、小静脉和小动脉内皮细胞的整体基因表达谱。R21 选择的格式是专门为探索性项目设计的，可能会涉及相当大的风险，但 导致可能对某一研究领域产生重大影响的新技术的发展。在这 因此，本研究的结果将为通过神经血管评估局部基因表达奠定基础。 单位(即，内皮细胞和密切相关的神经细胞)沿中枢微血管树在 健康和疾病。反过来，这些研究将使更精确的内皮模型的形成成为可能 脑微血管生理和病理生理过程的分子基础。