Quantitative proteomic analysis of the senescence secretory phenotype in UV-A induced senescent human corneal endothelial cells.

Abstract

Purpose : We presented that 5J/cm2 ultraviolet (UV) -A radiation-induced cellular senescence in human corneal endothelial cells (hCEnCs) (Numa et al. ARVO2023 in New Orleans). This study aims to investigate the senescence and the senescence secretory phenotype (SASP) in the UV-A-induced senescent hCEnCs via comprehensive protein coverage.

Methods : Primary hCEnCs were obtained from donor corneas. The hCEnCs were irradiated with 5J/cm² UV-A or 10Gy-IR, a more conventional method of senescence induction, and were cultured to undergo senescent. Conditioned media collected from senescent hCEnCs and quiescent controls. Isolated proteins were characterized, and mass spectrometry (LC-MS/MS) analysis was performed for comparative proteome profile of UV-A induced senescent hCEnCs compared to IR-induced senescent hCEnCs.

Results : This analysis identified 766 significantly altered quantifiable proteins (with at least 2 unique peptides) in the UV-A vs. control comparison, and 729 proteins in the IR vs. control comparison. Proteomics also revealed that 93.8% of the proteins, including SASPs such as CXCL1, CXCL8, MMP2, TGFB1, TGFB2, and GDF15, significantly upregulated in UV-A-induced senescent hCEnCs overlapped with those induced by IR. Gene ontology analysis revealed that UV-A-induced senescent hCEnCs exhibited elevated levels of proteins associated with endopeptidase regulator activity, collagen, extracellular matrix organization, and glycolysis pathway.

Conclusions : In this study, we determined that UV-A modulated the expression of most proteins typically altered upon IR treatment, and more specific pathways to corneal endothelial diseases than IR.

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