Molecular Mechanisms of Ischemia

描述（由申请人提供）： 该计划项目的中心主题是，强有力的内源性神经保护机制编码在基因组中，这些基因的一个子集的表达有助于确定细胞是否在缺血中存活。科学目标是识别和表征这些基因及其蛋白质产物运作的神经保护途径。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主动过程。鉴定内源性神经保护剂的基因产物将大大有助于我们理解缺血性神经元损伤的病理生理学，并为中风和相关疾病（如创伤性脑损伤）的新治疗方法指明方向。例如，发现一个转录因子和靶基因的网络，调节脑缺血耐受性，将推动药理学的努力，以模拟这种效果。我们将集中在体内和体外系统，其中内源性神经保护已被诱导和大脑已作出容忍随后的缺血性损伤（缺血预处理和耐受性）。发现缺血中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鉴定耐受中转录调节的基因（项目1）。然后将在缺血和耐受模型中对鉴定的基因进行体外研究，以表征和确认神经保护功能（项目2）。最后，将使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物（项目3）。基因组学核心（核心A）将为每个项目提供Affymbox微阵列分析。我们在太平洋西北国家实验室的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）将协调手稿、计算机连接、数据共享、演讲者旅行、赠款管理和相互作用实验室的统计咨询，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.