NHC Key Laboratory of Human Stem and Reproductive Engineering, School of Basic Medical Science, Central South University, Changsha, China.
Clinical Research Center for Reproduction and Genetics in Hunan Province, Reproductive and Genetic Hospital of Citic-Xiangya, Changsha, China.
Stem Cells Dev. 2023 Nov;32(21-22):681-692. doi: 10.1089/scd.2023.0034. Epub 2023 Oct 20.
Retinitis pigmentosa (RP) is a prevalent inherited retinal degenerative disease resulting from photoreceptor and pigment epithelial apoptosis. The is the most commonly associated pathogenic gene in RP. However, mutations (c.512C>T P171L) have been infrequently reported, and the RP pathogenesis caused by these mutations remains unclear. The objective of this study was to investigate the impact of (c.512C>T P171L) mutation on retinal cell differentiation and elucidate the underlying mechanisms of RP. An effective retinal organoid induction scheme for inhibiting the Wnt signaling pathway was selected for further experiments, and the established cell line HES-406 was demonstrated to be heterozygous for c.512C>T, with a normal karyotype and pluripotency potential. Furthermore, the development of HES-406 organoids may be delayed, and apoptosis detection and co-localization revealed that HES-406 organoids had more apoptotic cells than HES-90 in the outer nuclear layer (ONL), mutant RHO protein was mislocalized in the endoplasmic reticulum (ER), and stress-related and apoptotic gene expression increased. Overall, our study elucidated a possible mechanism by which ER stress caused by RHO P171L protein mislocalization may lead to ONL cell apoptosis.
色素性视网膜炎（RP）是一种常见的遗传性视网膜退行性疾病，由光感受器和色素上皮细胞凋亡引起。 是 RP 最常相关的致病基因。然而， 突变（c.512C>T P171L）报道较少，这些突变引起的 RP 发病机制尚不清楚。本研究旨在探讨 （c.512C>T P171L）突变对视网膜细胞分化的影响，并阐明 RP 的潜在机制。选择了一种有效的抑制 Wnt 信号通路的视网膜类器官诱导方案进行进一步实验，所建立的细胞系 HES-406 表现为 c.512C>T 的杂合子，具有正常的核型和多能性潜力。此外，HES-406 类器官的发育可能延迟，凋亡检测和共定位显示，HES-406 类器官的外核层（ONL）中凋亡细胞比 HES-90 多，突变 RHO 蛋白在内质网（ER）中定位错误，应激相关和凋亡基因表达增加。总的来说，我们的研究阐明了 RHO P171L 蛋白定位错误导致 ER 应激可能导致 ONL 细胞凋亡的可能机制。
