Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis

作者：Kang‐Chieh S Huang, Mong-Lien Wang, Shih-Jen Chen, Jean-Cheng Kuo, Won-Jing Wang, Phan Nguyen Nhi Nguyen, K. Wahlin, Jyh-Feng Lu, Audrey A. Tran, Michael Shi, Yueh Chien, A. Yarmishyn, P. Tsai, Tien-Chun Yang, W. Jane, Chia-Ching Chang, Chi-Hsien Peng, T. Schlaeger, S. Chiou · 发表于：Stem Cell Reports · 年份：2019 · DOI：10.1016/j.stemcr.2019.09.010 · 被引用次数：82 · 研究领域：Medicine、Biology

Summary X-linked juvenile retinoschisis (XLRS), linked to mutations in the RS1 gene, is a degenerative retinopathy with a retinal splitting phenotype. We generated human induced pluripotent stem cells (hiPSCs) from patients to study XLRS in a 3D retinal organoid in vitro differentiation system. This model recapitulates key features of XLRS including retinal splitting, defective retinoschisin production, outer-segment defects, abnormal paxillin turnover, and impaired ER-Golgi transportation. RS1 mutation also affects the development of photoreceptor sensory cilia and results in altered expression of other retinopathy-associated genes. CRISPR/Cas9 correction of the disease-associated C625T mutation normalizes the splitting phenotype, outer-segment defects, paxillin dynamics, ciliary marker expression, and transcriptome profiles. Likewise, mutating RS1 in control hiPSCs produces the disease-associated phenotypes. Finally, we show that the C625T mutation can be repaired precisely and efficiently using a base-editing approach. Taken together, our data establish 3D organoids as a valid disease model.