Master Student Sun Yat-sen University Guangzhou, Guangdong, China (People's Republic)
Abstract: Glaucoma is the most prevalent retinal degeneration disease that is caused by the lesion and loss of retinal ganglion cells (RGCs). Finding a method to regenerate RGCs represents a critical direction toward curing glaucoma, but there is still no effective one to regenerate RGCs. In our previous study, we identified a combination of three transcription factors (TFs)—Ascl1, Brn3b, and Isl1(abbreviated as ABI)—that efficiently reprogrammed fibroblasts into RGC-like neurons (iRGCs) in vitro. In this study, we used mouse late-stage retinal progenitor cells (late-RPCs) which lack the competence to differentiate into RGCs under the established developmental process, as a model to examine the ability of ABI to induce the RGC fate through direct somatic cell reprogramming in situ. Our results showed that overexpressing ABI reprogrammed the developmental competent state of the late RPCs, enabling them efficiently to generate RGCs. The ABI-induced RGCs exhibited RGC-like electrophysiological activities, projected axons to and formed synaptic connections with the lateral geniculate body (LGN) and superior colliculus (SC). Single cell RNA sequencing confirmed the RGC nature of the induced cells and revealed the molecular route underlying the reprogramming process. Furthermore, CUT&RUN and ATAC-Seq analyses revealed roles of each TF in the ABI combination during the reprogramming process. Taken together, our study demonstrated the potent capability of Ascl1, Brn3b, and Isl1 in in vivo RGC fate induction. These findings may pave the way for translational applications in glaucoma treatment in the future.
Funding Source: the National Natural Science Foundation of China (32470848)
