Mechanisms of Neuroprotection in Ischemic Tolerance in Mice

小鼠缺血耐受的神经保护机制

• 批准号: 6779019
• 负责人: Mary Stenzel-Pore
• 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/6779019
• 关键词: antisense nucleic acid; biological models; cell type; cerebral ischemia /hypoxia; cytoprotection; flow cytometry; functional /structural genomics; gene expression; gene targeting; genetic regulation; genetically modified animals; laboratory mouse; messenger RNA; microarray technology; neurogenetics; neuroregulation; osteopontin; polymerase chain reaction; prostaglandin endoperoxide synthase; protease inhibitor; protein structure function; secretory protein; serine proteinases; stimulus /response; stroke; vascular endothelial growth factors

lschemic tolerance is the genome's own endogenous protective response to ischemic stress. The goal of this application is to discover the genes and gene pathways regulating this protection and to do so with a goal of providing new therapeutic strategies for stroke. We will be guided by genes discovered in tolerance models developed in project 1 and genes further characterized as neuroprotective in vitro in project 2. We will then directly test Candidate gene products and candidate neuroprotective pathways in mice by use of routine models over-expressing a candidate gene, in knockouts or by altering the expression of the gene product of interest. We will use mouse lines available to us or create new models if required as we have done previously. Thus we plan to: 1) Determine whether increased expression or presence of candidate neuroprotective genes identified in Projects 1 and 2 using in vitro and in vivo models of preconditioning, improves neuronal survival in ischemia in vivo. 2) Determine whether decreased expression or inhibition of function of candidate neuroprotective genes, identified in Projects 1 and 2 using in vitro and in vivo models of preconditioning, reduces tolerance and neuronal survival in ischemia in vivo. 3) Determine which genes are altered in tolerance in the absence of neuroprotective genes to elucidate the function of novel genes in ischemic preconditioning. We will begin with a novel inflammatory pathway surrounding the osteopontin gene, which was revealed in our initial microarray screening. By a combined analysis of genes discovered in multiple models in vivo and in vitro in projects 1 and 2 our goal is to discover gene sets which share common promoter elements and reveal the common restricted pathways regulating tolerance to ischemia in brain. Knowledge of such pathways should be broadly relevant to injury states across organs and beyond ischemia.

缺血耐受是基因组自身对缺血应激的内源性保护反应。该应用的目标是发现调节这种保护的基因和基因途径，并以此为中风提供新的治疗策略。我们将以项目 1 开发的耐受模型中发现的基因以及进一步的基因为指导 在项目 2 中被表征为体外神经保护作用。然后，我们将通过使用过度表达候选基因的常规模型、敲除或通过改变感兴趣的基因产物的表达，直接在小鼠中测试候选基因产物和候选神经保护途径。我们将使用可用的鼠标线或根据需要创建新模型，就像我们之前所做的那样。因此我们计划： 1) 使用体外和体内预处理模型确定项目 1 和 2 中确定的候选神经保护基因的表达增加或存在是否改善体内缺血中的神经元存活。 2)确定项目1和2中使用体外和体内预处理模型确定的候选神经保护基因的表达减少或功能抑制是否会降低体内缺血时的耐受性和神经元存活。 3）确定在缺乏神经保护基因的情况下哪些基因的耐受性发生改变，以阐明新基因在缺血预处理中的功能。 我们将从围绕骨桥蛋白基因的一种新的炎症途径开始，这在我们最初的微阵列筛选中已被揭示。通过对项目 1 和 2 中多个体内和体外模型中发现的基因进行组合分析，我们的目标是发现具有共同启动子元件的基因集，并揭示调节脑缺血耐受性的共同限制途径。对这些途径的了解应该与跨器官和缺血以外的损伤状态广泛相关。