白内障是一种蛋白错误折叠疾病，自噬-溶酶体功能异常与之密切相关，但具体机制未明。TFEB是自噬-溶酶体系统（ALS）关键调控分子，可入核激活CLEAR网络基因表达，维持晶状体生理功能。申请人前期利用人iPSC体外再生晶状体（LB）诱导分化技术获得衰老LB和先天性白内障患者特异性突变LB，模拟年龄相关性及先天性白内障的晶状体病理改变。进而发现与正常LB相比，衰老和突变LB中TFEB表达水平下降，胞核分布减少，CLEAR网络ALS相关基因表达下降，自噬体降解受阻。据此提出假说：TFEB很可能通过CLEAR网络调控ALS功能影响晶状体内蛋白质稳态。本研究拟利用衰老及突变过表达细胞模型、衰老及突变LB模型、白内障小鼠模型、白内障患者晶状体样本，结合CRISPRi、DNA pull down、ChIP-seq、蛋白质谱等技术，探讨TFEB在白内障发生发展中的作用及机制，为白内障防治提供潜在药物靶点。

Cataract is a protein misfolding disease closely related to autophagy-lysosome dysfunction, yet the specific mechanism is largely unknown. TFEB is a key regulator of autophagy-lysosomal system (ALS), which translocate into the nucleus to activate the Coordinated Lysosomal Expression and Regulation (CLEAR) network and maintain the physiological functions of the lens. In our previous study, we developed a protocol to acquire regenerate lens (lentoid body, LB) from human induced pluripotent stem cells. Based on this, we obtained aged LB and congenital cataract patient-specific LB to simulate the pathological alterations of age-related cataract and congenital cataract. It was further discovered that the expression and nuclei distribution of TFEB were significantly decreased in aged LB and mutant LB, compared to normal LB. The expression of ALS-related genes in CLEAR network was decreased and autophagosome degradation was also hindered. Therefore, we proposed a hypothesis that TFEB is likely to regulate ALS function through mediating CLEAR network, thereby affecting lens proteostasis. This study aims to explore the role of TFEB in cataractogenesis and its specific mechanism, based on cell model, LB model, mouse model, and lens samples from cataract patients, through various techniques including CRISPRi, DNA pull down, co-immunoprecipitation, protein mass spectrometry, etc. This study is expected to provide potential therapeutic targets for cataract prevention and treatment in the future.