標題: Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis
作者: Huang, Kang-Chieh
Wang, Mong-Lien
Chen, Shih-Jen
Kuo, Jean-Cheng
Wang, Won-Jing
Phan Nguyen Nhi Nguyen
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
生物科技學系
Department of Biological Science and Technology
公開日期: 12-十一月-2019
摘要: 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.
URI: http://dx.doi.org/10.1016/j.stemcr.2019.09.010
http://hdl.handle.net/11536/153151
ISSN: 2213-6711
DOI: 10.1016/j.stemcr.2019.09.010
期刊: STEM CELL REPORTS
Volume: 13
Issue: 5
起始頁: 906
結束頁: 923
顯示於類別:期刊論文