Comparative Gene Expression in Human vs Rodent Stroke Models

项目摘要： 许多缺血性卒中临床试验的失败导致了越来越多的争论，即小鼠和大鼠是否 模型有效。我们认为，转化性卒中研究的关键不在于啮齿动物模型是否能够 完全预测人类中风的所有治疗方法，但如何正确使用细胞和动物模型进行药物治疗 发展，取决于目标机制。很可能当小鼠和大鼠模型 与人类中风相比，有些靶点和机制是相同的，有些是不同的。然而，在这方面， 没有系统的研究来评估小鼠和大鼠细胞和模型之间的相似性和差异， 与人类中风患者相比。这是我们试图填补的翻译中的关键空白。在这个项目中，我们将绘制 缺氧缺糖或脑缺血后神经元、星形胶质细胞和小胶质细胞的基因表达 从小鼠，大鼠和人类，并建立一个比较数据库的网络和途径反应，在所有 三个物种。 在目标1中，我们将检测来自小鼠、大鼠或大鼠的原代神经元、星形胶质细胞和小胶质细胞中的基因表达， 人在基线条件下和氧-葡萄糖剥夺后，通过基因阵列，并评估途径 使用标准的京都基因和基因组百科全书（KEGG）和基因本体（GO）分析。 将通过基于MetaBase和Incidity的z评分评估通路的上调或下调 数据库。在目标2中，我们将确定缺血周围神经元、星形胶质细胞和小胶质细胞中的基因反应， 青年与老年以及正常血压与高血压小鼠和大鼠局灶性脑缺血后皮质 模型与六名经过验证的人类中风患者进行比较。mRNA水平和基因通路反应 将再次用基因阵列和KEGG/GO分析进行评估。途径的上调或下调 将通过基于MetaBase和Incidity数据库的z评分进行评估。 本项目将建立一个神经元、星形胶质细胞和小胶质细胞相关基因的数据库 在体外对氧-葡萄糖剥夺和在体内对缺血作出反应的网络和途径。这 比较数据库将允许我们（a）比较啮齿动物与人类中风，（B）比较原代细胞培养 对比体内模型，和（c）比较年轻与老年动物，和（d）比较正常血压与高血压 动物最终，这个关系和交互式数据库应该允许用户询问潜在的 在细胞和啮齿动物中风模型中的翻译机制和靶点。

Project Summary: The failure of many clinical trials in ischemic stroke has led to increasing debate about whether mouse and rat models are valid. We propose that the key for translational stroke research is not whether rodent models can completely predict ALL treatments for human stroke, but how to correctly use cell and animal models for drug development, depending on the mechanisms being targeted. It is likely that when mouse and rat models are compared to human stroke, some targets and mechanisms are the same and some are different. However, there are no systematic studies to assess similarities and differences between mouse and rat cells and models vs human stroke patients. This is the critical gap in translation that we seek to fill. In this project, we will map gene expression after oxygen-glucose deprivation or cerebral ischemia in neurons, astrocytes and microglia from mouse, rats and humans, and build a comparative database of network and pathway responses in all three species. In Aim 1, we will detect gene expression in primary neurons, astrocytes and microglia from mouse, rat or human under baseline condition and after oxygen-glucose deprivation by gene array, and assess pathways with standard Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis. Upregulation or downregulation of pathways will be assessed via z-scores based on MetaBase and Ingenuity databases. In Aim 2, we will determine gene response in neurons, astrocytes and microglia in peri-ischemic cortex after focal cerebral ischemia in young vs aged as well as normotensive vs hypertensive mouse and rat models for comparison with six validated human stroke patients. mRNA levels and gene pathway responses will again be assessed with gene array and KEGG/GO analysis. Upregulation or downregulation of pathways will be assessed via z-scores based on MetaBase and Ingenuity databases. This project will produce a relational and interactive database of neuronal, astrocytic and microglial gene networks and pathways that respond to oxygen-glucose deprivation in vitro and ischemia in vivo. This comparative database will allow us to (a) compare rodents vs human stroke, (b) compare primary cell culture vs in vivo models, and (c) compare young vs aged animals, and (d) compare normotensive vs hypertensive animals. Ultimately, this relational and interactive database should allow users to interrogate potential mechanisms and targets for translation in cell and rodent stroke models.