NDUFA4基因缺陷通过调控VDAC1介导内质网应激及神经元凋亡参与Dandy-Walker畸形的作用机制研究

Dandy-Walker畸形（DWM）是主要累及小脑等后颅窝结构的常见先天性疾病之一。课题组前期研究已证实NDUFA4基因缺陷促进神经元凋亡并导致DWM的发生，但具体机制未明。预实验①成功构建了NDUFA4基因敲除鼠模型；②F2代敲除鼠出现小脑蚓部形态改变等DWM表型；③串联质谱分析发现敲除鼠线粒体膜蛋白VDAC1及内质网应激（ERS）标志物ATF4及CHOP表达上调；④在细胞模型中发现沉默NDUFA4可上调VDAC1并激活ERS凋亡通路，促进细胞凋亡。沉默NDUFA4同时干扰VDAC1表达可逆转ERS凋亡因子的相关改变，保护神经元。据此本项目提出科学假说：NDUFA4基因缺陷通过上调VDAC1介导ERS及神经元凋亡，最终导致DWM的发生。本课题拟进一步从细胞、分子、动物水平及正反面论证研究NDUFA4基因缺陷导致DWM的发病机制，为寻找神经保护治疗靶点提供科学依据。

Dandy-Walker malformation (DWM) is one of the most common congenital diseases, which mainly involves cerebellum and other posterior fossa structures. Previous studies have confirmed that NDUFA4 gene defect promotes neuronal apoptosis and leads to DWM, but the specific mechanism is not clear. Our previous research showed that: (1) NDUFA4 knockout mouse model was successfully constructed; (2) DWM phenotypes such as cerebellar vermis necrosis were found in F2 knockout mice; (3) the expression of VDAC1 and Endoplasmic reticulum stress (ERs) markers: ATF4 and CHOP were up-regulated in knockout mice by tandem mass spectrometry; (4) silencing NDUFA4 upregulated VDAC1 and activated ERs apoptosis pathway in cell model. Simutaneously silencing the expression of NDUFA4 and VDAC1 can reverse the related changes of ERs apoptotic factors and protect neurons. Based on these results, this project proposed a scientific hypothesis: NDUFA4 gene defect mediates ERs and neuronal apoptosis by up-regulating VDAC1, and eventually leads to DWM. This project intends to further study the pathogenesis of DWM caused by NDUFA4 gene defect from the cellular, molecular and animal level, so as to provide scientific basis for potential neuroprotective therapeutic targets.