Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis
Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis
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DOI：
10.1016/j.stemcr.2019.09.010
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发表时间：
2019-11-12
影响因子：
5.9
通讯作者：
Chiou, Shih-Hwa
中科院分区：
文献类型：
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作者：
Huang, Kang-Chieh;Wang, Mong-Lien;Chiou, Shih-Hwa
与RS 1基因突变相关的X连锁青少年视网膜劈裂症（XLRS）是一种具有视网膜劈裂表型的退行性视网膜病变。我们从患者产生人诱导多能干细胞（hiPSC）以在3D视网膜类器官体外分化系统中研究XLRS。该模型概括了XLRS的关键特征，包括视网膜分裂、有缺陷的视网膜分裂素产生、外节缺陷、异常桩蛋白周转和受损的ER-高尔基体运输。RS 1突变还影响感光细胞感觉纤毛的发育，并导致其他视网膜病变相关基因的表达改变。CRISPR/Cas9对疾病相关C625 T突变的校正使分裂表型、外节缺陷、桩蛋白动力学、纤毛标志物表达和转录组谱正常化。同样，在对照hiPSC中突变RS 1产生疾病相关表型。最后，我们证明了C625 T突变可以使用碱基编辑方法精确有效地修复。总之，我们的数据将3D类器官建立为有效的疾病模型。
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.