Abstract
Purpose :
Decreased trabecular meshwork (TM) cellularity is a critical pathogenic cause for primary open-angle glaucoma (POAG), yet therapies to regenerate the decellularized TM are very limited. Induced pluripotent stem cell-derived TM cells (iPSC-TM) can efficiently restore aqueous humor outflow and maintain intraocular pressure homeostasis. Here, advances in single-cell RNA-sequencing (scRNA-seq) provide answers to an important question: are there subpopulations of iPSC-TM cells that are specifically involved in regenerating damaged tissues?
Methods :
scRNA-seq was conducted to characterize the molecular mechanisms underlying TM regeneration. Intraocular pressure and outflow facility were measured by rebound tonometry and a custom-made syringe pump system. Immunohistochemistry (IHC), Western Blot, siRNA technique, and RT-PCR were employed to investigate the pro-proliferative effect of the selected subpopulation of iPSC-TM.
Results :
Our clustering analysis identified a group of alpha6 integrin-positive (ITGA6+) iPSC-TM with a distinct transcription pattern that was not observed among primary TM (pTM) cells. These ITGA6+ iPSC-TM not only stimulated cell proliferation of pTM much more efficiently than other iPSC-TM subtypes, but are also capable of restoring aqueous humor (AH) outflow in glaucoma. IHC results revealed that ITGA6+ iPSC-TM repopulated the TM and reversed damage to the inner wall of Schlemm’s canal. Finally, we unraveled the molecular mechanisms underlying the pro-proliferative effect of ITGA6+ iPSC-TM by IHC and siRNA technique, which is related to the robust expression of long non-coding RNA NEAT1.
Conclusions :
This study advances our understanding of the mechanism of iPSCs-based therapy and facilitates the development of a more efficient treatment for TM regeneration.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.