Neuroprotective Genes in Ischemic Tolerance

缺血耐受中的神经保护基因

• 批准号: 6779015
• 负责人: ROGER Pancoast SIMON
• 金额: $ 36.06万
• 依托单位: LEGACY EMANUEL HOSPITAL AND HEALTH CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6779015
• 关键词: apoptosis; cerebral ischemia /hypoxia; confocal scanning microscopy; cytoprotection; endotoxins; functional /structural genomics; gene expression; gene induction /repression; gene interaction; hippocampus; immunocytochemistry; in situ hybridization; informatics; laboratory mouse; laser capture microdissection; lipopolysaccharides; microarray technology; neurogenetics; neuroregulation; polymerase chain reaction; pyramidal cells; statistics /biometry; stimulus /response; stroke

Project 1: Focal ischemia affects gene expression in brain, and such gene expression affects outcome. Neuroprotective genes are induced by premonitory sublethal focal ischernia resulting in tolerance. Ischemic tolerance also results from sublethal stress caused by global ischemia and by systemic endotoxin priming. Our goal is to discover the subset of genesthat produce tolerance in each of these paradigms. Our previous approach to gene modulation in brain ischenaia was subtractive hybridization and cloning. We cloned 27 new genes over the past five years. We will now use microarray technology to increase our capabilities for gene discovery and focus the efforts on protective genes modeled in ischemic tolerance. We have developed new mouse models of tolerance in order to use murine gene chips and exploit the power of the mouse genome. Our approach is to use several models, each of which results in tolerance but with distinct stimuli or regional targets in brain. We will characterize tolerance to focal ischemia in cortex protected by prior brief focal ischemia or endotoxin priming, and will characterize tolerance to global ischemia in CA 1 neurons protected by prior brief global ischemia. Multiple models and replicates will permit sophisticated statistical analysis of data. Established core facilities for genomics will enable optimal data management and analysis. New informatics and data mining collaborators at Pacific Northwest National Laboratories Supercomputer and Bioinformatics divisions adds substantial strength to our data analysis capabilities. Our goal is to find endogenous pathways of neuroprotection modeled in tolerance. To this end we will identify genes and pathways regulated during tolerance by preconditioning focal ischemia (Aim 1); identify genes and pathways regulated during tolerance by endotoxin priming (Aim 2); and identify genes and pathways regulated during tolerance in the selectively vulnerable neurons of the CA 1 sector following global ischemia (Aim 3). Genes of known and unknown function discovered in these tolerance models will be advanced to projects two and three for validation in in vitro and in vivo systems based on a specific strategic plan.The elucidation of endogenous protective systems should identify novel approaches for stroke treatment.

项目1：局灶性缺血会影响大脑中的基因表达，这种基因表达会影响预后。神经保护基因是由前局部局灶性缺口诱导的，导致耐受性。缺血性耐受性也来自全球性缺血和全身性内毒素启动引起的全部压力。我们的目标是发现基因的子集在每个范式中产生耐受性。我们先前在脑部伊希纳氏菌中调节基因的方法是减法杂交和克隆。在过去的五年中，我们克隆了27个新基因。现在，我们将使用微阵列技术来提高基因发现的能力，并将精力集中在缺血性耐受性的保护基因上。我们开发了新的耐受性小鼠模型，以便使用鼠基因芯片并利用小鼠基因组的功率。我们的方法是使用多种模型，每种模型都会产生耐受性，但在大脑中具有不同的刺激或区域目标。我们将表征受到先前短暂局灶性缺血或内毒素启动保护的皮质局部缺血的耐受性，并将表征受到先前短暂全局缺血保护的CA 1神经元中对全局缺血的耐受性。多个模型和重复将允许对数据进行复杂的统计分析。 建立的基因组学核心设施将实现最佳数据管理和分析。西北太平洋国家实验室超级计算机和生物信息学部门的新信息和数据挖掘合作者为我们的数据分析功能增添了很大的优势。我们的目标是找到以耐受性建模的神经保护的内源性途径。为此，我们将通过预处理局灶性缺血来鉴定在耐受性期间调节的基因和途径（AIM 1）；通过内毒素启动确定在耐受性期间调节的基因和途径（AIM 2）；并确定在有选择性的公差期间调节的基因和途径 全球缺血后，CA 1部门的脆弱神经元（AIM 3）。在这些耐受性模型中发现的已知和未知功能的基因将基于特定的战略计划中的第二和第三项目进行验证。内源性保护系统的阐明应确定中风治疗的新方法。