利用iPSC-视网膜类器官研究4-HNE诱导Caspase-3凋亡通路在RDH12相关视网膜变性中的作用

视黄醇脱氢酶(RDH12)基因突变可导致严重的视网膜退行性疾病，发病机制不清。RDH12视网膜病变尚无理想的疾病模型，缺乏有效的治疗手段。我们拟构建RDH12突变iPSC-视网膜类器官，摸索合适的培养条件，进行光感受器细胞定向分化。以视网膜类器官研究对象，阐明患者特异RDH12突变的致病性和致病机制，包括突变对酶活性影响和细胞凋亡等的差异，以期发现新的分子靶点和通路。并通过对4-HNE加合物测定、Caspase-3信号通路关键蛋白测定、氧化应激指标检测、4-HNE拮抗剂作用、细胞凋亡等方面研究RDH12在4-HNE对光感受器毒性作用中的保护机制。本项目预期结果能够为RDH12视网膜病变的发病机制提供组织细胞学及氧化应激方面依据，为进一步的治疗研究提供新的靶点。

Mutations of the photoreceptor retinol dehydrogenase 12 (RDH12) gene cause severe retinal degeneration by mechanisms not completely resolved. There is no ideal disease model or cure up to now. The project aims to establish patient specific iPSC derived retinal organoids and to explore the culture conditions for the differentiation of photoreceptor cells. Based on the iPSC derived retinal organoids, we try to expore the the underlying pathogenic mechanisms of RDH12 retinopathy, including the effect of mutation on enzyme activity and cell apoptosis, leading to discovery of new targets for therapy and new pathways. By testing the 4-HNE adduct, key proteins of Caspase-3 pathway, oxidative stress markers, 4-HNE antagonist and cell apoptosis, the protective mechanism of RDH12 in detoxifying 4-HNE in photoreceptor cells will be studied. The expected results of this project will bring insight into the understanding of pathogenesis of RDH12-related recessive retinopathy in the aspect of cytohistology and oxidative stress, which then will lead to discovery of new targets for therapy and new pathways for further investigation.