Wolfram综合征（WS）是一种常染色体隐性遗传疾病，WS神经病变主要由神经元和神经胶质细胞的Wolframin蛋白基因（WFS1）突变所致。然而由于缺乏有效的研究手段，人WS神经病变发病机制尚不明确。我们利用CRISPR/Cas9靶向基因编辑技术建立了WFS1基因编辑的人多能干细胞等基因细胞系，并将其定向诱导分化为神经元和星型胶质细胞。前期研究发现WFS1敲除神经元树突短且分支少，而突触形成较多；WFS1敲除星型胶质细胞显著抑制神经元轴突生长。故推测：WFS1可能通过调控神经元及星型胶质细胞的形态和功能进而诱导WS神经病变。本研究拟通过前期建立的人等基因细胞系构建、神经细胞诱导分化、RNAseq基因表达谱分析、小分子化合物筛选等技术，阐明WFS1突变或敲除导致的神经元及星型胶质细胞形态和功能异常的分子机制，为WS神经病变发病机制的研究提供新思路，并为WS神经病变的治疗提供新药物。

Wolfram Syndrome (WS) is a rare autosomal recessive genetic disorder that leads to neuropathy by Wolframin protein (WFS1) gene mutations in neurons and glial cells. However, due to the lack of effective research models, the pathogenesis and mechanism of human WS neuropathy are still unclear. Here, we establish isogenic human pluripotent stem cells carrying WFS1 mutations using CRISPR/Cas9 gene-editing system. Using this system, we find that WS-derived neurons possess shorter total dendrites, increased numbers of spines and synapses; while WS-derived astrocytes significantly inhibit neuronal axon growth. In this project, we will identify detailed WFS1 genotype-specific defects of neurons or astrocytes by comparing WFS1 mutate/knock-out isogenic human pluripotent cells with wild-type controls and unveil the molecular mechanism under these defects. Finally, with our novel isogenic human pluripotent cell system, we will set up a high-throughput drug-screening platform targeting the defects of neural cells and try to rescue the neuropathy in WS. Taken together, our project will not only improve our understanding of the mechanism of WS neuropathy but also provide new pharmaceutical drug targets for the treatment of WS.