Otsuka Yuki, Imamura Keiko, Oishi Akio, Kondo Takayuki, Suga Mika, Yada Yuichiro, Shibukawa Ran, Okanishi Yasue, Sagara Yukako, Tsukita Kayoko, Tsujikawa Akitaka, Inoue Haruhisa
iPSC-based Drug Discovery and Development Team, RIKEN BioResource Research Center, Kyoto, Japan.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
iScience. 2022 Mar 8;25(4):103987. doi: 10.1016/j.isci.2022.103987. eCollection 2022 Apr 15.
Retinal dystrophies (RDs) lead to irreversible vision impairment with no radical treatment. Although photoreceptor cells (PRCs) differentiated from human induced pluripotent stem cells (iPSCs) are essential for the study of RDs as a scalable source, current differentiation methods for PRCs require multiple steps. To address these issues, we developed a method to generate PRCs from human iPSCs by introducing the transcription factors, CRX and NEUROD1. This approach enabled us to generate induced photoreceptor-like cells (iPRCs) expressing PRC markers. Single-cell RNA sequencing revealed the transcriptome of iPRCs in which the genes associated with phototransduction were expressed. Generated iPRCs exhibited their functional properties in calcium imaging. Furthermore, light-induced damage on iPRCs was inhibited by an antioxidant compound. This simple approach would facilitate the availability of materials for PRC-related research and provide a useful application for disease modeling and drug discovery.
视网膜营养不良（RDs）会导致不可逆的视力损害，且没有根治方法。尽管从人类诱导多能干细胞（iPSCs）分化而来的光感受器细胞（PRCs）作为一种可扩展的来源，对RDs的研究至关重要，但目前PRCs的分化方法需要多个步骤。为了解决这些问题，我们开发了一种通过引入转录因子CRX和NEUROD1从人类iPSCs生成PRCs的方法。这种方法使我们能够生成表达PRC标志物的诱导性光感受器样细胞（iPRCs）。单细胞RNA测序揭示了iPRCs的转录组，其中与光转导相关的基因得以表达。生成的iPRCs在钙成像中表现出其功能特性。此外，一种抗氧化化合物可抑制光对iPRCs的损伤。这种简单的方法将有助于提供与PRC相关研究的材料，并为疾病建模和药物发现提供有用的应用。
