The TCF4 gene regulates apoptosis of corneal endothelial cells in Fuchs endothelial corneal dystrophy

Nakagawa, Tatsuya, Honda, Tetsuro, Yuasa, Taichi, Nishiuchi, Go, Sato, Masakazu, Tokunaga, Ayumi, Nakahara, Makiko, Tourtas, Theofilos, Schlötzer-Schrehardt, Ursula, Kruse, Friedrich, Padmanabhan, Prema, Chatterjee, Amit, Sathe, Gajanan, Ghose, Vivek, Janakiraman, Narayanan, Blake, Derek J. ORCID: https://orcid.org/0000-0002-5005-4731, Koizumi, Noriko, Elchuri, Sailaja and Okumura, Naoki 2025. The TCF4 gene regulates apoptosis of corneal endothelial cells in Fuchs endothelial corneal dystrophy. Investigative Ophthalmology & Visual Science 66 (3) , 16. 10.1167/iovs.66.3.16

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Abstract

Purpose: Fuchs endothelial corneal dystrophy (FECD) is a progressive corneal disorder characterized by excessive extracellular matrix (ECM) accumulation and corneal endothelial cell death. CTG trinucleotide repeat expansion in the transcription factor 4 (TCF4) gene represents the most significant genetic risk factor. This study aimed to elucidate the role of TCF4 in FECD pathogenesis through comprehensive proteomic analysis. Methods: Corneal endothelial cells isolated from patients with FECD harboring TCF4 trinucleotide repeat expansion were immortalized to establish an FECD cell model (iFECD). CRISPR/Cas9-mediated genome editing was employed to generate TCF4-knockout iFECD cells. Whole-cell proteome analysis was performed using liquid chromatography–mass spectrometry, followed by pathway enrichment analysis of differentially expressed proteins (DEPs). The effects of TCF4 deletion on TGF-β–mediated protein aggregation and cell death were evaluated using Western blot analysis, flow cytometry, and aggresome detection assays. Results: Proteomic analysis identified 88 DEPs among 6510 detected proteins. Pathway analysis revealed significant enrichment in ECM-associated pathways, oxidative stress responses, and cellular motility. TCF4 deletion attenuated TGF-β–induced cell death in iFECD cells. Concordantly, Western blot analysis demonstrated that TCF4 deletion suppressed TGF-β2–mediated cleavage of caspase-3 and poly (ADP-ribose) polymerase. Flow cytometric analysis of Annexin V–positive cells confirmed reduced apoptosis in TCF4-deleted cells following TGF-β2 treatment. Additionally, aggresome detection assays revealed that TCF4 deletion diminished TGF-β2–induced protein aggregation. Conclusions: This study demonstrates a crucial role for TCF4 in FECD pathogenesis, particularly in ECM regulation and protein aggregation–induced cell death.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Schools > Medicine
Publisher:	Association for Research in Vision and Ophthalmology
ISSN:	0146-0404
Date of First Compliant Deposit:	19 March 2025
Date of Acceptance:	11 February 2025
Last Modified:	19 Mar 2025 10:00
URI:	https://orca.cardiff.ac.uk/id/eprint/176986

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