Atg5-Mediated Lipophagy Induces Ferroptosis in Corneal Epithelial Cells in Dry Eye Disease

作者：Xin Zuo, Hao Zeng, Xue Yang, Dalian He, Bowen Wang, Jin Yuan · 发表于：Investigative Ophthalmology & Visual Science · 年份：2024 · DOI：10.1167/iovs.65.14.12 · 被引用次数：20 · 研究领域：Ferroptosis and cancer prognosis、Inflammasome and immune disorders、interferon and immune responses

Purpose: Ferroptosis occurred in corneal epithelial cells has been implicated in the inflammation in dry eye disease (DED). Given the proposed link between ferroptosis and autophagy, this study aims to investigate the role of autophagy in driving ferroptosis in corneal epithelial cell and enrich the pathogenesis underlying DED. Methods: DED models were established in C57BL/6 mice via scopolamine injection and in human corneal epithelial cell line (HCEC) using hyperosmotic medium. Lipidomic and transcriptomic analysis were conducted to assess lipid metabolism and regulatory pathways. Atg5 expression was manipulated in vivo using cholesterol-modified small interfering RNA. Lipid droplets (LDs) and lysosomes were labeled with BODIPY 493/503 and Lysotracker Red DND-99, respectively. Western blot, immunofluorescence (IF) staining, co-immunoprecipitation (CO-IP), transmission electron microscopy and microplate reader were used to explore protein expressions and interactions, cellular structures, and free fatty acid (FFA) content. Results: Our results revealed that autophagy was activated in DED, as evidenced by lipidomic and transcriptomic analyses. Enhanced lipophagy was observed in HCECs exposed to hyperosmolarity, manifested by lysosome-LD co-localization and autophagic vacuoles containing LDs. Upregulation of Atg5 promoted lipophagy, leading to elevated cellular FFA levels, lipid peroxidation, and expression of ferroptosis markers. Interaction between Atg5 and perilipin3 was co...