NOX- Mediated oxidative Stress Creates a Fibrotic and Inflammatory Milieu In Vernal Keratoconjunctivitis (VKC)

Abstract

Purpose :

Purpose: Vernal keratoconjunctivitis (VKC) is a persistent and severe allergic eye disease that primarily affects children, often resulting in severe ocular complications with the potential for blindness. Here we report for the first time the role of Nicotinamide Adenine Dinucleotide Phosphate Oxidase (NOX)-mediated oxidative stress as the key driver of VKC pathogenesis.

Methods : Methods: Unstimulated tears were collected from VKC patients for proteomic analysis using LC-MS. Olink’s Proximity Extension Assay (PEA) technology was employed for further cytokine analysis, due to its high sensitivity for low-abundance cytokines and its 1 μl of sample volume requirement. Human corneal limbal epithelial cells (HCLE) were treated to for the functional validation of the proteomics findings and evaluation of reactive oxygen species (ROS) by DCFDA-based real-time PCR (RTPCR) was used to evaluate the gene expression in VKC patient-derived tissue samples.

Results : Results: LC-MS-based proteomic analysis showed the role of NOX and oxidative stress pathways. Olink’s PEA-based platform detected an increased expression of matrix metalloproteases (MMP12, MMP1, 9) and pro-inflammatory cytokines with high sensitivity (. HCLE cells treated with VKC patient-derived tears showed increased inflammatory markers, ROS, and NOX -2, -5 levels.

Conclusions : Conclusion: The current medicine regime for VKC patients effectively decreases inflammation but not fibrosis. NOX-mediated oxidative stress could be a reason for tissue damage and the adult onset of the disease or ocular complications like Glaucoma, cataracts, and keratoconus. Targeting NOX pathway strategy to prevent VKC pathogenesis.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.