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

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.

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.

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.

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.