Single- cell transcriptomics reveals maturation of transplanted stem cell-derived retinal pigment epithelial cells toward native state

Transplantation of stem cell-derived retinal pigment epithelial (RPE) cells is considered a viable therapeutic option for age-related macular degeneration (AMD). Several landmark Phase I/II clinical trials have demonstrated safety and tolerability of RPE transplants in AMD patients, albeit with limited efficacy. Currently, there is limited understanding of how the recipient retina regulates the survival, maturation, and fate specification of transplanted RPE cells. To address this, we transplanted stem cell-derived RPE into the subretinal space of immunocompetent rabbits for 1 mo and conducted single -cell RNA sequencing analyses on the explanted RPE monolayers, compared to their age-matched in vitro counterparts. We observed an unequivocal retention of RPE identity, and a trajectory-inferred survival of all in vitro RPE populations after transplantation. Furthermore, there was a unidirectional maturation toward the native adult human RPE state in all transplanted RPE, regardless of stem cell resource. Gene regulatory network analysis suggests that tripartite transcription factors (FOS, JUND, and MAFF) may be specifically activated in posttransplanted RPE cells, to regulate canonical RPE signature gene expression crucial for supporting host photoreceptor function, and to regulate prosurvival genes required for transplanted RPE's adaptation to the host subretinal microenvironment. These findings shed insights into the transcriptional landscape of RPE cells after subretinal transplantation, with important implications for cell -based therapy for AMD.

Automated summary

Transplantation of stem cell-derived retinal pigment epithelial (RPE) cells is a potential treatment for age-related macular degeneration (AMD). However, there is limited understanding of how the recipient retina regulates the survival and maturation of transplanted RPE cells. In this study, we transplanted stem cell-derived RPE into rabbits and analyzed their gene expression, finding that the transplanted RPE cells retained their identity and showed maturation towards the normal RPE state. Transcription factors FOS, JUND, and MAFF were activated in the transplanted RPE cells and played important roles in regulating gene expression related to supporting host photoreceptor function and promoting cell survival.