Direct reprogramming has been widely explored to generate various types of neurons for neurobiological research and translational medicine applications, but there is still no efficient reprogramming method to generate retinal ganglion cell (RGC)-like neurons, which are the sole projection neurons in the retina. Here, we show that three transcription factors, Ascl1, Brn3b, and Isl1, efficiently convert fibroblasts into RGC-like neurons (iRGCs). Furthermore, we show that the competence of cells to enter iRGC reprogramming route is determined by the cell-cycle status at a very early stage of the process. The iRGC reprogramming route involves intermediate states that are characterized by a transient inflammatory-like response followed by active epigenomic and transcriptional modifications. Our study provides an efficient method to generate iRGCs, which would be a valuable cell source for potential glaucoma cell replacement therapy and drug screening studies, and reveals the key cellular events that govern successful neuronal fate reprogramming.

直接重编程已被广泛探索以生成用于神经生物学研究和转化医学应用的各种类型的神经元，但仍然没有有效的重编程方法来生成视网膜神经节细胞 (RGC) 样神经元，这是视网膜中唯一的投射神经元。在这里，我们展示了三个转录因子Ascl1、Brn3b和Isl1, 有效地将成纤维细胞转化为 RGC 样神经元 (iRGC)。此外，我们表明细胞进入 iRGC 重编程途径的能力是由该过程早期阶段的细胞周期状态决定的。iRGC 重编程途径涉及中间状态，其特征是短暂的炎症样反应，然后是活跃的表观基因组和转录修饰。我们的研究提供了一种生成 iRGC 的有效方法，这将是潜在的青光眼细胞替代疗法和药物筛选研究的宝贵细胞来源，并揭示了控制成功神经元命运重编程的关键细胞事件。