TMEM166基因在氧、糖剥夺培养神经细胞和基因改造小鼠脑缺血模型中的整合生物学研究

TMEM166为北京大学人类疾病基因研究中心通过高通量功能筛选获得的新基因，最近我们的实验证明TMEM166调控大鼠局部脑缺血后神经细胞的自噬和凋亡（Experimental Neurology，2012）。由此我们凝练出TMEM166如何调控缺血和缺氧性神经细胞死亡的作用机理这一科学问题。本研究项目在分子、细胞、组织、器官、系统和机体多层次上开展整合生物学研究，研究内容有三个部分：（1）通过将TMEM166基因转染培养海马神经细胞的过表达和去功能化，研究TMEM166调节神经细胞自噬和凋亡的机理；（2）在氧、糖剥夺培养的海马神经细胞分别采用自噬和凋亡的多种特异性抑制剂和基因干扰，研究TMEM166基因调节缺氧诱导自噬和凋亡的分子传导通道；（3）分别采用TMEM166转基因和基因敲除小鼠的局部脑缺血模型，深入研究TMEM166对缺血性脑损伤的作用机制，探讨新的临床治疗策略。

On the bases of the finding on new gene of TMEM166 by Peking University Center for Human Disease Genomics,and our new publication that TMEM166 regelating the autophagy and apoptosis following the focal ischemia in rat(Experimental Neurology 2012[Epub ahead of print].The mechanism of TMEM166 on the regulation in the central neuroanl system following brain ischemia and hypoxia is the subject of this research program. We will study in the fild of cells,tissues,organs,neural system and in vivo,which including three parts:(1)To study the mechanism of TMEM166 on the autophagy and apoptosis by translating TMEM166 gene over-expression and dysfunction in the culture of hippcampus neurons in vitro;(2)To study the molecular signaling pathway of TMEM166 regulating the autohagy and apoptosis by using different inhibitors of autohagy and apoptosis in the culture of hippcampus neurons in vitro;(3) To study the mechanism and effection of TMEM166 in vivo by using TMEM166 gene transgenic mouse and gene knock out mouse by genetic manipulation suffered middle cerebral artery occlusion, to explore the possible strategy for clinical therapy in cerebral strock by inhibiting the TMEM166 signaling pathway of autophagy and apoptosis.