Necroptosis Driven by JAK/STAT-RIPK-MLKL Signaling Mediates Corneal Epithelial Erosion.

PURPOSE: Recurrent corneal erosion (RCE) is an ocular surface disease with poor epithelial-stromal adhesion and limited therapies. This study aimed to elucidate the role of necroptosis, a pro-inflammatory form of programmed cell death, in the pathogenesis of corneal erosion.

METHODS

An in vitro UVB injury model was established in human corneal epithelial cells (HCEpiCs), followed by unbiased LC-MS/MS proteomics and pathway enrichment analysis. In parallel, a UVB-induced corneal erosion model was generated in C57BL/6 mice. To test therapeutic relevance, mice received a topical neutralizing anti-IFN-γ antibody or vehicle after injury. Corneal epithelial integrity and stromal architecture were assessed histologically, while activation of IFN-γ-JAK/STAT signaling, RIPK1/RIPK3/MLKL necroptosis, DAMP release, NLRP3 inflammasome assembly, and IL-1β maturation were examined using immunofluorescence, qRT-PCR, and Western blotting.

RESULTS

Proteomic profiling showed significant enrichment of necroptosis-related proteins following UVB exposure. UVB-treated corneal epithelium exhibited increased IFN-γ signaling and JAK/STAT activation, accompanied by elevated expression and activation of RIPK1, RIPK3, and MLKL. Necroptosis was associated with increased release of DAMPs, heightened NLRP3 inflammasome activation, and increased maturation/secretion of IL-1β. In vivo, topical anti-IFN-γ treatment reduced corneal epithelial defects, improved epithelial-stromal attachment, and decreased activation markers of the RIPK-MLKL and NLRP3-IL-1β pathways.

CONCLUSIONS

Our findings unveil a novel and critical pathogenic axis in corneal erosion, where IFN-γ signaling drives a necroptotic-inflammatory loop. This work provides a new molecular framework for understanding corneal epithelial injury and identifies the IFN-γ-necroptosis axis as a promising therapeutic target for RCE.