Long-term corneal recovery by simultaneous delivery of hPSC-derived corneal endothelial precursors and nicotinamide

Abstract

Human pluripotent stem cells (hPSCs) hold great promise for the treatment of various human diseases. However, their therapeutic benefits and mechanisms for treating corneal endothelial dysfunction remain undefined. Here, we developed a therapeutic regimen consisting of the combination of hPSC-derived corneal endothelial precursors (CEPs) with nicotinamide (NAM) for effective treatment of corneal endothelial dysfunction. In rabbit and nonhuman primate models, intracameral injection of CEPs and NAM achieved long-term recovery of corneal clarity and thickness, similar with the therapeutic outcome of cultured human corneal endothelial cells (CECs). The transplanted human CEPs exhibited structural and functional integration with host resident CECs. However, the long-term recovery relied on the stimulation of endogenous endothelial regeneration in rabbits, but predominantly on the replacing function of transplanted cells during the 3-year follow-up in nonhuman primates, which resemble human corneal endothelium with limited regenerative capacity. Mechanistically, NAM ensured in vivo proper maturation of transplanted CEPs into functional CECs by preventing premature senescence and endothelial-mesenchymal transition within the TGF-β–enriched aqueous humor. Together, we provide compelling experimental evidence and mechanistic insights of simultaneous delivery of CEPs and NAM as a potential approach for treating corneal endothelial dysfunction.

Recent advances in cell-based regenerative therapies for corneal disease

“…Meanwhile, additional candidates for adjunctive therapy are being investigated, such as fibroblast grow factor 1 derivatives which may promote endothelial proliferation and migration [61,62], and other preclinical studies use cells that will not require corneal donation, such as human SC from embryonic [63], induced pluripotent [64–67], umbilical cord-derived [68], dental pulp [69] and eyelid hair follicle-derived [70] sources. None of these approaches are currently in human trials.…”

Cell therapy in corneal endothelial disease

“…In the last 18 months however, several preclinical animal trials have been published. We highlight the interesting contributions of four, one with human embryonic stem cell (hESC)-derived corneal endothelial precursor (CEP) cells, two using human-induced pluripotent cell (hiPSC)-derived CEC and one with donor cadaveric human CECs [15 ▪ –18 ▪ ].…”

“…Li et al [18 ▪ ] cultured hESC-derived precursor cells for injection in a rabbit and monkey model of bullous keratopathy. hESCs first underwent a 5-day induction to neural crest cells (NCCs), exhibiting cobblestone morphology and neural crest markers (P75, HNK-1, AP-2α and AP-2β).…”

Corneal Endothelial Characteristics in Normal Chinese Han Children and Youngsters: A Study from the Specular Microscopy Descriptions

“…In vivo, postpartum human CECs are terminally differentiated (TD), nonmitotic organs, which are static in the G1 phase of the cell cycle [14]. A plurality of factors, including cellcell contact inhibition, the exits of cell cycle negative regulators (e.g., cell cyclin-dependent kinase inhibitors, p15INK4b, and p27kip1), growth inhibitors (e.g., transforming growth factor-β and TGF-β) immersing in the aqueous humor, and stress-induced precocious senescence, contribute to the retention of CECs in their nonregenerative conformation [15,16].…”