A novel protein encoded by hsa_circ_0068626 contributes to age-related cataract via the p62/Keap1/Nrf2 signaling pathway-mediated ferroptosis

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research Pub Date : 2025-09-03 DOI:10.1016/j.exer.2025.110601

Yuxin Chen , Huajian Li , Lei Wu , Xi Zou , Yue Zhang , Rongrong Huang , Yong Wang

{"title":"A novel protein encoded by hsa_circ_0068626 contributes to age-related cataract via the p62/Keap1/Nrf2 signaling pathway-mediated ferroptosis","authors":"Yuxin Chen , Huajian Li , Lei Wu , Xi Zou , Yue Zhang , Rongrong Huang , Yong Wang","doi":"10.1016/j.exer.2025.110601","DOIUrl":null,"url":null,"abstract":"<div><div>This study investigates whether circular RNAs (circRNAs) modulate ferroptosis in lens epithelial cells (LECs) during age-related cataract (ARC) pathogenesis via novel encoded proteins. Initial circRNA-sequencing identified hsa_circ_0068626 (circTFRC) as significantly upregulated in ARC, predominantly localized to the cytoplasm through nuclear-cytoplasmic fractionation and fluorescence in situ hybridization (FISH). Functional assays revealed that circTFRC depletion impaired LECs proliferation and viability, while overexpression exacerbated ferroptosis, evidenced by elevated intracellular reactive oxygen species (ROS) and Fe<sup>2+</sup> level via fluorescence probes and flow cytometry. Mechanistically, circTFRC harbored an open reading frame (ORF) and internal ribosome entry site (IRES), enabling translation of the circTFRC-236aa protein, confirmed by polysome profiling and custom antibody detection. Western blot analyses demonstrated that circTFRC-236aa activated the p62/Keap1/Nrf2 axis, correlating with GPX4 suppression and ferroptosis. Transmission electron microscopy further visualized mitochondrial morphological abnormalities consistent with ferroptotic stress. Collectively, these findings establish circTFRC as a pro-ferroptotic regulator in ARC, where its encoded circTFRC-236aa drives pathological progression via p62/Keap1/Nrf2 pathway activation, offering a novel therapeutic target for mitigating ARC-associated LECs damage.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"260 ","pages":"Article 110601"},"PeriodicalIF":2.7000,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental eye research","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0014483525003720","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

This study investigates whether circular RNAs (circRNAs) modulate ferroptosis in lens epithelial cells (LECs) during age-related cataract (ARC) pathogenesis via novel encoded proteins. Initial circRNA-sequencing identified hsa_circ_0068626 (circTFRC) as significantly upregulated in ARC, predominantly localized to the cytoplasm through nuclear-cytoplasmic fractionation and fluorescence in situ hybridization (FISH). Functional assays revealed that circTFRC depletion impaired LECs proliferation and viability, while overexpression exacerbated ferroptosis, evidenced by elevated intracellular reactive oxygen species (ROS) and Fe²⁺ level via fluorescence probes and flow cytometry. Mechanistically, circTFRC harbored an open reading frame (ORF) and internal ribosome entry site (IRES), enabling translation of the circTFRC-236aa protein, confirmed by polysome profiling and custom antibody detection. Western blot analyses demonstrated that circTFRC-236aa activated the p62/Keap1/Nrf2 axis, correlating with GPX4 suppression and ferroptosis. Transmission electron microscopy further visualized mitochondrial morphological abnormalities consistent with ferroptotic stress. Collectively, these findings establish circTFRC as a pro-ferroptotic regulator in ARC, where its encoded circTFRC-236aa drives pathological progression via p62/Keap1/Nrf2 pathway activation, offering a novel therapeutic target for mitigating ARC-associated LECs damage.

查看原文本刊更多论文

一种由hsa_circ_0068626编码的新蛋白通过p62/Keap1/Nrf2信号通路介导的铁下沉参与年龄相关性白内障。

本研究探讨环状rna （circRNAs）是否通过新的编码蛋白在年龄相关性白内障（ARC）发病过程中调节晶状体上皮细胞（LECs）中的铁下垂。初始circrna测序发现hsa_circ_0068626 （circTFRC）在ARC中显著上调，通过核细胞质分离和荧光原位杂交（FISH）主要定位于细胞质。功能分析显示，circTFRC缺失会损害LECs的增殖和活力，而过表达会加剧铁凋亡，荧光探针和流式细胞术显示细胞内活性氧（ROS）和Fe2+水平升高。从机制上讲，circTFRC具有开放阅读框（ORF）和内部核糖体进入位点（IRES），能够翻译circTFRC-236aa蛋白，这已被多体分析和定制抗体检测证实。Western blot分析表明，circTFRC-236aa激活了p62/Keap1/Nrf2轴，与GPX4抑制和铁凋亡相关。透射电镜进一步显示线粒体形态异常与铁致应力一致。综上所述，这些发现表明circTFRC在ARC中是一种亲铁性调节因子，其编码的circTFRC-236aa通过p62/Keap1/Nrf2通路激活驱动病理进展，为减轻ARC相关lec损伤提供了一种新的治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Experimental eye research 医学-眼科学

CiteScore

6.80

自引率

5.90%

发文量

323

审稿时长

66 days

期刊介绍： The primary goal of Experimental Eye Research is to publish original research papers on all aspects of experimental biology of the eye and ocular tissues that seek to define the mechanisms of normal function and/or disease. Studies of ocular tissues that encompass the disciplines of cell biology, developmental biology, genetics, molecular biology, physiology, biochemistry, biophysics, immunology or microbiology are most welcomed. Manuscripts that are purely clinical or in a surgical area of ophthalmology are not appropriate for submission to Experimental Eye Research and if received will be returned without review.