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 |