Iron-Sulfur Clusters and Iron Responsive Element Binding Proteins Mediate Iron Accumulation in Corneal Endothelial Cells in Fuchs Dystrophy.

IF 5 2区 医学 Q1 OPHTHALMOLOGY
Emma M Hartness, Hanna Shevalye, Jessica M Skeie, Timothy Eggleston, Matthew G Field, Gregory A Schmidt, Pornpoj Phruttiwanichakun, Aliasger K Salem, Mark A Greiner
{"title":"Iron-Sulfur Clusters and Iron Responsive Element Binding Proteins Mediate Iron Accumulation in Corneal Endothelial Cells in Fuchs Dystrophy.","authors":"Emma M Hartness, Hanna Shevalye, Jessica M Skeie, Timothy Eggleston, Matthew G Field, Gregory A Schmidt, Pornpoj Phruttiwanichakun, Aliasger K Salem, Mark A Greiner","doi":"10.1167/iovs.66.4.23","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Evidence suggests that corneal endothelial cell (CEC) death in Fuchs endothelial corneal dystrophy (FECD) is due to ferroptosis, an iron-mediated cell death. Iron-sulfur cluster (ISC)-containing aconitases and the iron responsive element binding proteins IREBP1 and IREBP2 are known mediators of iron homeostasis. This study investigates mechanisms underlying iron dysregulation in CECs and proposes a role for ISCs and IREBPs in the context of FECD pathogenesis.</p><p><strong>Methods: </strong>We studied gene expression of proteins responsible for ISC synthesis and iron homeostasis in human and mouse CECs and analyzed published RNA sequencing datasets. We validated a subset of transcriptional changes between FECD and control tissues using microfluidic Western blotting with human CEC tissues. Finally, we silenced proteins involved in ISC synthesis or iron homeostasis in cell cultures and assessed ferroptosis susceptibility.</p><p><strong>Results: </strong>RNA-seq and qPCR data demonstrated significantly decreased transcription of genes required for ISC synthesis in FECD tissues (P < 0.05). Protein quantification revealed a significant decrease in mitochondrial aconitase (P < 0.05), ferredoxin 1 (P < 0.001), and mitofusin (P < 0.05), and a significant increase in cysteine desulfurase (P < 0.05), cytosolic aconitase/IREBP1, and IREBP2 (P < 0.05) in FECD tissues. Silencing studies revealed increased susceptibility to ferroptosis upon siRNA knockdown of ferredoxin 1 (P < 0.05).</p><p><strong>Conclusions: </strong>We identified differential gene expression of proteins responsible for ISC synthesis, ISC-containing proteins, IREBPs that mediate cellular iron homeostasis, and mitofusin, which promotes mitochondrial fusion in FECD. We also identified increased susceptibility to ferroptosis after ferredoxin 1 knockdown in CECs. These results advance an ISC- and IREBP-mediated mechanism of iron accumulation in FECD CECs.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 4","pages":"23"},"PeriodicalIF":5.0000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Investigative ophthalmology & visual science","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1167/iovs.66.4.23","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

Purpose: Evidence suggests that corneal endothelial cell (CEC) death in Fuchs endothelial corneal dystrophy (FECD) is due to ferroptosis, an iron-mediated cell death. Iron-sulfur cluster (ISC)-containing aconitases and the iron responsive element binding proteins IREBP1 and IREBP2 are known mediators of iron homeostasis. This study investigates mechanisms underlying iron dysregulation in CECs and proposes a role for ISCs and IREBPs in the context of FECD pathogenesis.

Methods: We studied gene expression of proteins responsible for ISC synthesis and iron homeostasis in human and mouse CECs and analyzed published RNA sequencing datasets. We validated a subset of transcriptional changes between FECD and control tissues using microfluidic Western blotting with human CEC tissues. Finally, we silenced proteins involved in ISC synthesis or iron homeostasis in cell cultures and assessed ferroptosis susceptibility.

Results: RNA-seq and qPCR data demonstrated significantly decreased transcription of genes required for ISC synthesis in FECD tissues (P < 0.05). Protein quantification revealed a significant decrease in mitochondrial aconitase (P < 0.05), ferredoxin 1 (P < 0.001), and mitofusin (P < 0.05), and a significant increase in cysteine desulfurase (P < 0.05), cytosolic aconitase/IREBP1, and IREBP2 (P < 0.05) in FECD tissues. Silencing studies revealed increased susceptibility to ferroptosis upon siRNA knockdown of ferredoxin 1 (P < 0.05).

Conclusions: We identified differential gene expression of proteins responsible for ISC synthesis, ISC-containing proteins, IREBPs that mediate cellular iron homeostasis, and mitofusin, which promotes mitochondrial fusion in FECD. We also identified increased susceptibility to ferroptosis after ferredoxin 1 knockdown in CECs. These results advance an ISC- and IREBP-mediated mechanism of iron accumulation in FECD CECs.

铁硫簇和铁响应元件结合蛋白介导富克斯营养不良患者角膜内皮细胞中的铁积累。
目的:有证据表明,福氏内皮性角膜营养不良症(FECD)中角膜内皮细胞(CEC)的死亡是由于铁突变(一种铁介导的细胞死亡)引起的。含铁硫簇(ISC)的乌头酶和铁应答元件结合蛋白 IREBP1 和 IREBP2 是已知的铁平衡介质。本研究探讨了CECs中铁失调的机制,并提出了ISCs和IREBPs在FECD发病机制中的作用:我们研究了人和小鼠 CECs 中负责 ISC 合成和铁稳态的蛋白质的基因表达,并分析了已发表的 RNA 测序数据集。我们利用微流体Western印迹法对人CEC组织与FECD和对照组织之间的转录变化进行了验证。最后,我们沉默了细胞培养物中参与 ISC 合成或铁稳态的蛋白质,并评估了铁中毒的易感性:结果:RNA-seq 和 qPCR 数据显示,在 FECD 组织中,ISC 合成所需的基因转录量明显减少(P < 0.05)。蛋白质定量显示,在 FECD 组织中,线粒体乌头酶(P < 0.05)、铁氧还蛋白 1(P < 0.001)和丝裂霉素(Mitofusin)明显减少,而半胱氨酸脱硫酶(P < 0.05)、细胞质乌头酶/IREBP1 和 IREBP2(P < 0.05)则明显增加。沉默研究显示,在 siRNA 敲除铁氧还蛋白 1 后,铁中毒的易感性增加(P < 0.05):结论:我们发现在 FECD 中,负责 ISC 合成的蛋白、含 ISC 的蛋白、介导细胞铁稳态的 IREBPs 和促进线粒体融合的 mitofusin 的基因表达存在差异。我们还发现,在 CECs 中敲除铁氧还蛋白 1 后,其对铁突变的易感性增加。这些结果推进了由 ISC 和 IREBP 介导的 FECD CECs 铁积累机制。
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
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信