Pou2f2 enhances retinal ganglion cell survival and axon regeneration after optic nerve injury

Retinal ganglion cell (RGC) degeneration and the limited regenerative capacity of mammalian central nervous system axons result in irreversible vision loss in optic neuropathies. Identifying intrinsic regulators that promote both neuroprotection and axon regeneration is critical for developing effective therapies. This study identified POU domain class 2 transcription factor 2 (Pou2f2) as a previously unrecognized regulator involved in RGC biology. Single-cell RNA sequencing analysis of developing mouse and human retinas showed that Pou2f2 is enriched in RGC clusters and suppressed by an RGC fate suppressor in the mouse retinal progenitor cell culture. Pou2f2 protein was detected in mouse RGCs and optic nerves, where its expression decreased following optic nerve crush (ONC). In primary RGC cultures, AAV2-mediated Pou2f2 overexpression enhanced neurite outgrowth, whereas Pou2f2 knockdown reduced neurite length. In vivo, intravitreal injection of Pou2f2 overexpression AAV2s increased axon regeneration, improved RGC survival, and partially preserved RGC function after ONC. Bulk RNA sequencing data revealed early activation of cytoskeletal growth pathways and later upregulation of DNA repair-associated genes, whereas immunostaining and in vitro analyses suggested activation of AKT/mTOR signaling following Pou2f2 overexpression. Together, these findings identify Pou2f2 as a promising molecular target for gene-based therapies for RGC degenerative diseases, including glaucoma and other optic neuropathies.