A Balancing Act in Corneal Epithelial Repair: A MAPK-JUN/EGR1/TFAP2A Network Regulates Ferroptotic Cell Fate

Hanyi Jiang,1,* Zhiwei Chen,2,* Wenkang Luan,3,* Jia Li,4 Ningbei Yin1 1Department of Cleft Lip and Palate, Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 2The Third Medical Aesthetic Center, Plastic Surgery Hospital (Institute), Beijing, People’s Republic of China; 3Department of Auricular Reconstruction, Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 4Department of Ophthalmology, Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jia Li, Department of Ophthalmology, Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences & Peking Union Medical College, No. 33 Ba-Da-Chu Road, Shi Jing Shan District, Beijing, 100144, People’s Republic of China, Email [email protected] Ningbei Yin, Department of Cleft Lip and Palate, Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences & Peking Union Medical College, No. 33 Ba-Da-Chu Road, Shi Jing Shan District, Beijing, 100144, People’s Republic of China, Tel +86-10-53968006, Email [email protected]: To explore the role of ferroptosis in the process of corneal epithelial repair and to elucidate the underlying molecular regulatory mechanisms.Methods: This study performed transcriptomic analysis based on the zebrafish corneal epithelial repair dataset (GSE193784). We intersected differentially expressed genes with the core weighted gene co-expression network analysis (WGCNA) module, and integrated Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, protein-protein interaction (PPI) network construction, and cross-referencing with the FerrDb v2 database to screen for key ferroptosis-related molecules involved in the repair process. Subsequently, using an Erastin-induced ferroptosis model in human corneal epithelial cells (HCE-T), we performed in vitro validation via RT-qPCR. Additionally, we compared the identified transcriptomic signatures with the latest mammalian and human single-cell atlases to assess cross-species conservation.Results: A total of 252 differentially expressed genes (DEGs) were identified, Intersection with the antiquewhite1 WGCNA module, which is most highly correlated with corneal epithelial repair, yielded 99 overlapping genes. Functional enrichment analysis revealed their significant roles in transcriptomic reprogramming, with a prominent enrichment in the MAPK signaling pathway. Further cross-screening using a PPI network and the FerrDb v2 database pinpointed three core biological factors: jun, egr1, and tfap2a. In vitro experimental verification revealed their differential expression patterns of JUN downregulation, EGR1 upregulation and TFAP2A downregulation under ferroptosis stress. Cross-species bioinformatic comparisons demonstrated that this MAPK-driven transcriptomic reprogramming is highly consistent with the hyper-proliferative and highly plastic cellular states observed during mammalian and human corneal repair.Conclusion: This study found that JUN, EGR1 and TFAP2A, the core biological factors in the corneal epithelial repair process, are closely related to ferroptosis. This suggests that key biological factors enriched in the MAPK pathway may affect corneal epithelial repair by regulating ferroptosis.Keywords: corneal epithelial injury, ferroptosis, bioinformatics analysis, hub genes