新生儿缺血缺氧性脑病（HIE）严重威胁新生儿健康，但发病机制不明。申请人前期研究显示心脏骤停-心肺复苏（CA-CPR）诱导的新生动物HIE模型在处理6-24h内出现大脑自噬流受损，并发现自噬流受损与CHMP2A/VPS4介导的自噬溶酶体关闭异常密切相关。故提出研究假说：在CA-CPR 诱导的HIE中，CHMP2A在损伤神经元中表达降低，抑制VPS4介导的自噬溶酶体关闭及成熟，导致自噬流受损和HIE发生。本项目拟通过免疫电镜等技术，在CA-CPR 诱导的HIE模型中首先阐明CHMP2A/VPS4介导的自噬溶酶体关闭及成熟的分子机制，其次验证该过程是否会受到Beclin-1调控，最后在体水平探索CHMP2A/VPS4基因干预是否能恢复受损自噬流，缓解神经元损伤。本项目将揭示CHMP2A/VPS4介导自噬溶酶体的关闭及成熟在HIE中的机制，将为HIE防治及药物开发提供科学依据，具有重要临床意义。

Neonatal hypoxic ischemic encephalopathy (HIE) is a serious threat to the physical health of newborns, but its pathogenesis is still unclear. The previous studies have shown that the autophagic flux was impaired within 6 to 24 h after restoration of spontaneous circulation (ROSC) in the brain of neonatal animal HIE model induced by cardiac arrest - cardiopulmonary resuscitation (CA - CPR), which is consistent with the second and early third phases in clinical patients suffered from HIE. Moreover, through series of preliminary experiments, we focused on CHMP2A-mediated autolysosome closure and maturation. Therefore, a hypothesis is proposed: in the CA-CPR induced neonatal HIE mice model, the expression of CHMP2A is decreased in injured neurons, which leads to VPS4-mediated impairment of autolysosome closure and maturation. This project intends to investigate the mechanism of CHMP2A/VPS4-mediated autolysosome closure and maturation in CA-CPR-induced HIE model by using immunoelectron microscopy and other molecular biology technology. Further we will verify whether the process is regulated by Beclin-1 and explore whether CHMP2A and VPS4 gene intervention can restore autophagic flux and alleviate neuronal damage in vivo. Through this project, we will unveil the mechanism of CHMP2A/VPS4-mediated autolysosome closure and maturation in CA-CPR-induced HIE. It will provide a scientific basis for HIE prevention and drug development, which has important clinical significance.