Restoration of corneal endothelial dysfunction by transplanting functional corneal endothelial cells (CEC) derived from human induced pluripotent stem cells

Abstract

Purpose : To develop a highly efficient method for induction of CECs from cord blood-originated iPSCs and investigate the efficacy and safety of transplantation of iPSC-derived CECs into a corneal endothelial dysfunction (CED) rabbit model.

Methods : Cord blood-derived iPSC (generated in a clinical grade) was used to maintain and differentiate into CEC. For CEC differentiation, direct differentiation (iPSC-CEC; ACE2) or iPSC-neural crest cell (NCC)-CEC differentiation (ACE1) were performed. Single cell RNA-sequeuncing was performed using the 10X Genomics platform. A total of four different RNA libraries were prepared. Immunocytochemistry and Western blotting were performed. New Zealand white rabbits were used for iPSC-derived CEC transplantation. We injected iPSC-CECs (1×10⁶ iPSC-CECs in 150 μl of PBS supplemented with 100 μM of Fasudil) into the anterior chamber and followed for up to 36 weeks.

Results : iPSC-derived CECs exhibited a well-preserved hexagonality and expressed a group of CEC-specific markers. Single cell sequencing data showed similar functionalities of iPSC-CEC as compared to primary human CEC in terms of ATP synthesis and metabolism, cell adhesion, migration, extracellular vesicles, developmental process, anatomical structure development, and serine/threonine kinase activity. Our in vivo study demonstrated safety and therapeutic efficacy of iPSC-CEC on a CED rabbit model, as the transplanted group showed improved levels of corneal transparency. No immune rejections and inflammatory responses were detected in the host corneas.

Conclusions : Functional CECs were successfully induced from hiPSC using an improved protocol of differentiation. Noticeable recovery of corneal clarity of the CED model without responses of graft rejection indicates therapeutic capacity of iPSC-CEC as a powerful source of clinical therapy for patients with CED.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.