巨自噬/自噬失调与多种神经系统疾病有关，包括维西综合征。我们的目的是确定自噬在早期大脑发育中的作用。我们从人类胚胎干细胞中产生神经元，并通过在 EPG5 基因中引入功能丧失突变来开发 Vici 综合征模型。自噬相关基因在神经祖细胞阶段上调。在神经元祖细胞阶段用巴弗洛霉素 A1 处理抑制自溶酶体形成，延迟神经元分化。值得注意的是，WNT（Wnt 家族成员）信号传导可能是潜在机制的一部分，该机制受到自噬介导的 DVL2（蓬乱片段极性蛋白 2）降解的负调控。自溶酶体形成的破坏可能导致 WNT 信号传导失败，从而延迟神经元分化。 EPG5突变扰乱了自溶酶体的形成，随后诱导类器官中祖细胞分化和皮质层生成的缺陷。自噬破坏会导致类器官变小，重现维西综合征相关的小头畸形，并验证我们研究的疾病相关性。缩写：BafA1：巴弗洛霉素 A1； co-IP：免疫共沉淀； DVL2：蓬乱片段极性蛋白2； EPG5：异位P-颗粒5自噬束缚因子； gRNA，引导RNA； hESC：人类胚胎干细胞； KO：淘汰赛； mDA，中脑多巴胺； NIM：神经诱导介质； NPC：神经元祖细胞； qPCR：定量聚合酶链式反应； UPS：泛素-蛋白酶体系统； WNT：Wnt家庭成员； WT：野生型。

Macroautophagy/autophagy dysregulation is associated with various neurological diseases, including Vici syndrome. We aimed to determine the role of autophagy in early brain development. We generated neurons from human embryonic stem cells and developed a Vici syndrome model by introducing a loss-of-function mutation in the EPG5 gene. Autophagy-related genes were upregulated at the neuronal progenitor cell stage. Inhibition of autolysosome formation with bafilomycin A1 treatment at the neuronal progenitor cell stage delayed neuronal differentiation. Notably, WNT (Wnt family member) signaling may be part of the underlying mechanism, which is negatively regulated by autophagy-mediated DVL2 (disheveled segment polarity protein 2) degradation. Disruption of autolysosome formation may lead to failure in the downregulation of WNT signaling, delaying neuronal differentiation. EPG5 mutations disturbed autolysosome formation, subsequently inducing defects in progenitor cell differentiation and cortical layer generation in organoids. Disrupted autophagy leads to smaller organoids, recapitulating Vici syndrome-associated microcephaly, and validating the disease relevance of our study.Abbreviations: BafA1: bafilomycin A1; co-IP: co-immunoprecipitation; DVL2: dishevelled segment polarity protein 2; EPG5: ectopic P-granules 5 autophagy tethering factor; gRNA, guide RNA; hESC: human embryonic stem cells; KO: knockout; mDA, midbrain dopamine; NIM: neural induction media; NPC: neuronal progenitor cell; qPCR: quantitative polymerase chain reaction; UPS: ubiquitin-proteasome system; WNT: Wnt family member; WT: wild type.