Autosomal dominant optic atrophy (ADOA), mostly caused by heterozygous OPA1 mutations and characterized by retinal ganglion cell (RGC) loss and optic nerve degeneration, is one of the most common types of inherited optic neuropathies. Previous work using a two-dimensional (2D) differentiation model of induced pluripotent stem cells (iPSCs) has investigated ADOA pathogenesis but failed to agree on the effect of OPA1 mutations on RGC differentiation. Here, we use 3D retinal organoids capable of mimicking in vivo retinal development to resolve the issue. We generated isogenic iPSCs carrying the hotspot OPA1 c.2708_2711delTTAG mutation and found that the mutant variant caused defective initial and terminal differentiation and abnormal electrophysiological properties of organoid-derived RGCs. Moreover, this variant inhibits progenitor proliferation and results in mitochondrial dysfunction. These data demonstrate that retinal organoids coupled with gene editing serve as a powerful tool to definitively identify disease-related phenotypes and provide valuable resources to further investigate ADOA pathogenesis and screen for ADOA therapeutics.

常染色体显性视神经萎缩（ADOA）主要由OPA1杂合突变引起，以视网膜神经节细胞（RGC）丢失和视神经变性为特征，是最常见的遗传性视神经病类型之一。先前的工作使用诱导多能干细胞 (iPSC) 的二维 (2D) 分化模型研究了 ADOA 发病机制，但未能就OPA1突变对 RGC 分化的影响达成一致。在这里，我们使用能够模仿体内视网膜发育的 3D 视网膜类器官来解决这个问题。我们生成了携带热点OPA1 c.2708_2711delTTAG 突变的同基因 iPSC，发现该突变体导致类器官来源的 RGC 的初始和终末分化缺陷以及电生理特性异常。此外，这种变异会抑制祖细胞增殖并导致线粒体功能障碍。这些数据表明，视网膜类器官与基因编辑相结合，可以作为明确识别疾病相关表型的强大工具，并为进一步研究 ADOA 发病机制和筛选 ADOA 治疗方法提供宝贵的资源。