转录网络调控的恒河猴iPSC分化及青光眼视神经损伤的替代治疗研究

由细胞重编程获得的多潜能干细胞（iPSC），对青光眼等视网膜神经变性疾病的神经再生具有重要价值。我们先前报道通过调控Math5基因表达，诱导小鼠iPSC定向分化，形成具有长突触的RGC样细胞；本项目在以往研究基础上，结合RGCs转录调控网络，通过调控Wnt，BMP与Nodal通路诱导恒河猴iPSC分化为视网膜祖细胞，进而调控RGCs相关基因包括Pax6，Atoh7和Notch信号通路，诱导iPSC定向分化为有功能的视网膜神经节样细胞。并结合细胞衰老相关信号通路，监测并抑制分化过程中的P53表达和线粒体氧化应激，分析抗衰老对iPSC来源的RGCs的体外生长维持作用。最终我们将iPSC来源的高效分化的RGC样细胞，进行恒河猴青光眼动物模型视网膜下腔注射，监测其体内视神经再生的可能性及效率，为探索青光眼患者自体神经节细胞替代治疗提供理论和实验基础。

Induced pluripotent stem cells (iPSCs)derived from somatic cell reprogramming, have important value in neural regeneration of glaucoma and other retinal neurodegenerative diseases. We previously reported that mouse iPSC can directly differentiate into RGC-like cells by manipulation of Math5 expression and other signaling pathways. Based on the previous studies,this proposal will take advantage of the RGCs transcription regulation network, induce rhesus iPSC to differentiate into retinal progenitor cells by regulation of the Wnt, BMP and Nodal pathway. To directly differentiate rhesus iPSC into functional retinal ganglion-like cells, we will regulation of RGCs-related genes, including Pax6, Atoh7 and Notch signaling pathway. During the differentiation process, We will investigate cellular senescence signaling pathway, and suppress P53 expression and mitochondiral oxidative stress to maintain the growth of iPSC derived RGCs. Finally, we will inject the RGC-like cells efficiently differentiated from iPSC, into the subretina of rhesus monkey with experimental glaucoma, and monitor the possibility and efficiency of neural regenration. Our research will provide a theoretical and experimental basis for exploring ganglion cell replacement therapy for glaucoma patients.