The hiPSC-derived corneal endothelial progenitor-like cell recovers the rabbit model of corneal endothelial dystrophy

Yi-Jen Hsueh; Hung-Chi Chen; Yu-Yun Pan; Fang-Chi Hsiao; Shun-Jie Yang; Mei-Chun Liu; Wei-Yu Lai; Guigang Li; David Hui-Kang Ma; Yaa-Jyuhn James Meir

doi:10.1016/j.jare.2024.05.008

The hiPSC-derived corneal endothelial progenitor-like cell recovers the rabbit model of corneal endothelial dystrophy

J Adv Res. 2024 May 8:S2090-1232(24)00184-X. doi: 10.1016/j.jare.2024.05.008. Online ahead of print.

Authors

Affiliations

¹ Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Dept. of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan.
² Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Dept. of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan.
³ Dept. of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan.
⁴ Dept. of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan.
⁵ Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan.
⁶ Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei Province 430030, China.
⁷ Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Dept. of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Department of Chinese Medicine, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan.
⁸ Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan; Dept. of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan; Dept. of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33305, Taiwan. Electronic address: jmeir@mail.cgu.edu.tw.

PMID: 38729560
DOI: 10.1016/j.jare.2024.05.008

Abstract

Introduction: Corneal endothelial dysfunction results in cornea opacity, damaging sightedness, and affecting quality of life. A corneal transplant is the current effective intervention. Due to the scarcity of donated cornea, such an unmet medical need requires a novel therapeutic modality.

Objectives: Customizing patients' corneal endothelial progenitor cells with proliferative activity and lineage restriction properties shall offer sufficient therapeutic cells for corneal endothelial dystrophy.

Methods: The customized induced human corneal endothelial progenitor-like cell (iHCEPLC) was obtained through cell fate conversions starting from PBMC (peripheral blood mononuclear cell), hiPSC (human induced pluripotent stem cell), and hNCC (human neural crest cell), while it finally reached the iHCEPLC state via a series of induction. Several molecular diagnoses were applied to depict its progenitor state, including RNAseq, FlowCytometer, immunostainings, and rtPCR. Significantly, it can be induced to gain differentiation maturity through contact inhibition. In addition, a BAK-mediated rabbit model of corneal endothelial dystrophy was established in the present study to test the therapeutic effectiveness of the iHCEPLC.

Results: After inducing cell fate conversion, the specific HCEC markers were detected by rtPCR and immunostaining in iHCEPLC. Further, RNAseq was applied to distinguish its progenitor-like cell fate from primary human corneal endothelial cells (HECE). FlowCytometry profiled the heterogeneity subpopulation, consistently displaying a subtle difference from primary HCEC. A terminal differentiation can be induced in iHCEPLC, addressing its progenitor-like fate. iHCEPLC can restore the BAK-based rabbit model of corneal endothelial dystrophy. Immunohistochemistry verified that such acuity restoration of the BAK-treated cornea is due to the introduced iHCEPLC, and such therapeutic effectiveness is observed in the long term.

Conclusion: Here, we demonstrated that customized iHCEPLC has long-term therapeutic efficacy. As a progenitor cell, our iHCEPLC has a restricted cell lineage nature and can proliferate in vitro, supporting sufficient therapeutic candidate cells. Due to the immune-privileged nature of the cornea, our iHCEPLC proves the principle of therapeutical feasibility in both autogenic and allogeneic modalities.

Keywords: Human corneal endothelial cell (HCEC); Human corneal endothelial progenitor-like cells (HCEPLC); Human induced pluripotent stem cell (hiPSC); Neural crest cell (NCC); Peripheral blood mononuclear cell (PBMC).