Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, MSC0610, 6 Center Drive, Bethesda, MD 20892, USA.
Ocular Gene Therapy Core, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Stem Cell Reports. 2021 Feb 9;16(2):252-263. doi: 10.1016/j.stemcr.2020.12.018. Epub 2021 Jan 28.
Mutations in the photoreceptor transcription factor gene cone-rod homeobox (CRX) lead to distinct retinopathy phenotypes, including early-onset vision impairment in dominant Leber congenital amaurosis (LCA). Using induced pluripotent stem cells (iPSCs) from a patient with CRX-I138fs48 mutation, we established an in vitro model of CRX-LCA in retinal organoids that showed defective photoreceptor maturation by histology and gene profiling, with diminished expression of visual opsins. Adeno-associated virus (AAV)-mediated CRX gene augmentation therapy partially restored photoreceptor phenotype and expression of phototransduction-related genes as determined by single-cell RNA-sequencing. Retinal organoids derived from iPSCs of a second dominant CRX-LCA patient carrying K88N mutation revealed the loss of opsin expression as a common phenotype, which was alleviated by AAV-mediated augmentation of CRX. Our studies provide a proof-of-concept for developing gene therapy of dominant CRX-LCA and other CRX retinopathies.
CRX 基因（cone-rod homeobox）中的突变会导致不同的视网膜病变表型，包括显性莱伯先天性黑矇（LCA）的早期视力损伤。我们利用一位 CRX-I138fs48 突变患者的诱导多能干细胞（iPSC），在视网膜类器官中建立了 CRX-LCA 的体外模型，该模型通过组织学和基因谱分析显示感光细胞成熟缺陷，视蛋白表达减少。腺相关病毒（AAV）介导的 CRX 基因增强治疗部分恢复了感光细胞表型和光转导相关基因的表达，这是通过单细胞 RNA 测序确定的。源自第二位携带 K88N 突变的显性 CRX-LCA 患者 iPSC 的视网膜类器官表现出视蛋白表达缺失的共同表型，而 AAV 介导的 CRX 增强可以缓解这种情况。我们的研究为开发显性 CRX-LCA 和其他 CRX 视网膜病变的基因治疗提供了概念验证。
