Browsing by Author "Chang, Chia-Ching"

Now showing 1 - 1 of 1
  • Thumbnail Image
    Item
    Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis
    (Elsevier, 2019-11-12) Huang, Kang-Chieh; Wang, Mong-Lien; Chen, Shih-Jen; Kuo, Jean-Cheng; Wang, Won-Jing; Nguyen, Phan Nguyen Nhi; Wahlin, Karl J.; Lu, Jyh-Feng; Tran, Audrey A.; Shi, Michael; Chien, Yueh; Yarmishyn, Aliaksandr A.; Tsai, Ping-Hsing; Yang, Tien-Chun; Jane, Wann-Neng; Chang, Chia-Ching; Peng, Chi-Hsien; Schlaeger, Thorsten M.; Chiou, Shih-Hwa; Biology, School of Science
    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.