Molecular Mechanisms of Ischemia

描述(申请人提供)：该计划项目的中心主题是，强大的内源性神经保护机制在基因组中编码，这些基因的子集的表达有助于确定细胞是否能在缺血中存活。科学目标是识别和表征这些基因以及它们的蛋白质产物发挥作用的神经保护途径。这种方法的基本原理是理解大脑对损伤的反应是一个涉及新蛋白质合成的活跃过程。识别内源性神经保护剂的基因产物将有助于我们理解缺血性神经元损伤的病理生理学，并将为中风和相关疾病(如创伤性脑损伤)的新治疗方法指明方向。例如，调控脑缺血耐受的转录因子和靶基因网络的发现将推动模拟这一效应的药理学努力。我们将集中于体内和体外系统，在这些系统中，内源性神经保护已经被诱导，并且大脑已经对随后的缺血损伤(缺血预适应和耐受)进行了耐受。在缺血中发现神经保护基因的策略是使用缺血耐受的小鼠模型和微阵列分析来识别在耐受中转录调控的基因(项目1)。然后，将在缺血和耐受模型中体外研究已识别的基因，以确定和确认神经保护功能(项目2)。最后，将利用药理学和遗传学方法，通过增加或减少其在小鼠体内的表达来研究具有神经保护作用的基因产品(项目3)。基因组学核心(核心A)将为每个项目提供Affymetrix微阵列分析。我们在太平洋西北国家实验室超级计算机和生物信息学部门的合作者将通过条件概率方法对基因簇进行网络分析，并使用序列相似性分析对未知基因进行功能分配。行政核心(核心B)将协调相互作用的实验室的手稿、计算机连接、数据共享、演讲者差旅、赠款管理和统计咨询，以及通过内部和外部咨询委员会进行的科学咨询。

DESCRIPTION (provided by applicant): The central theme of this program project is that potent endogenous mechanisms of neuroprotection are encoded in the genome and that the expression of a subset of these genes helps to determine whether cells survive ischemia. The scientific goals are to identify and characterize these genes and the neuroprotective pathways through which their protein products operate. The rationale for this approach is the understanding that the brain's response to injury is an active process that involves new protein synthesis. Identifying gene products that are endogenous neuroprotectants would contribute significantly to our understanding of the pathophysiology of ischemic neuronal injury and would point the way toward new therapeutic approaches to stroke and to related disorders, such as traumatic brain injury. For example, the discovery of a network of transcription factors and target genes that regulate ischemic tolerance in brain would advance pharmacologic efforts to mimic this effect. We will focus on in vivo and in vitro systems wherein endogenous neuroprotection has been induced and the brain has been made tolerant to subsequent ischemic injury (ischemic preconditioning and tolerance). The strategy for discovering neuroprotective genes in ischemia is to use mouse models of ischemic tolerance and microarray analysis to identify genes that are transcriptionally regulated in tolerance (Project 1). Identified genes will then be studied in vitro in models of ischemia and tolerance to characterize and confirm neuroprotective function (Project 2). Finally, gene products that are neuroprotective will be investigated by increasing or reducing their expression in mice in vivo, using pharmacologic and genetic approaches (Project 3). A Genomics Core (Core A) will provide Affymetrix microarray analysis to each project. Our collaborators at Pacific Northwest National Laboratory's Supercomputer and Bioinformatics Division will employ network analysis of gene clusters via conditional probability approaches and functional assignment of unknown genes using analysis of sequence similarities. The Administrative Core (Core B) will coordinate manuscripts, computer connections, data sharing, speaker travel, grants management, and statistical consultation for the interacting laboratories, as well as scientific consultation through internal and external advisory boards.