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

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

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

Journal article

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

Emma M Hartness, Hanna Shevalye, Jessica M Skeie, Timothy Eggleston, Matthew G Field, Gregory A Schmidt, Pornpoj Phruttiwanichakun, Aliasger K Salem and Mark A Greiner

Investigative ophthalmology & visual science, Vol.66(4), p.23

04/01/2025

DOI: 10.1167/iovs.66.4.23

PMCID: PMC11993131

PMID: 40202733

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Abstract

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. 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. 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). 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.

Aconitate Hydratase - genetics

Aconitate Hydratase - metabolism

Animals

Blotting, Western

Cells, Cultured

Endothelium, Corneal - metabolism

Endothelium, Corneal - pathology

Ferroptosis

Fuchs' Endothelial Dystrophy - genetics

Fuchs' Endothelial Dystrophy - metabolism

Fuchs' Endothelial Dystrophy - pathology

Gene Expression Regulation - physiology

Homeostasis

Humans

Iron - metabolism

Iron-Sulfur Proteins - genetics

Iron-Sulfur Proteins - metabolism

Mice

Mice, Inbred C57BL

Details

Title: Subtitle: Iron-Sulfur Clusters and Iron Responsive Element Binding Proteins Mediate Iron Accumulation in Corneal Endothelial Cells in Fuchs Dystrophy
Creators: Emma M Hartness - University of Iowa
Hanna Shevalye - Lions Eye Bank (Coralville)
Jessica M Skeie - Lions Eye Bank (Coralville)
Timothy Eggleston - Lions Eye Bank (Coralville)
Matthew G Field - Minnesota Eye Consultants
Gregory A Schmidt - Lions Eye Bank (Coralville)
Pornpoj Phruttiwanichakun - University of Iowa
Aliasger K Salem - University of Iowa
Mark A Greiner - University of Iowa
Resource Type: Journal article
Publication Details: Investigative ophthalmology & visual science, Vol.66(4), p.23
DOI: 10.1167/iovs.66.4.23
PMID: 40202733
PMCID: PMC11993131
NLM abbreviation: Invest Ophthalmol Vis Sci
ISSN: 1552-5783
eISSN: 1552-5783
Publisher: ASSOC RESEARCH VISION OPHTHALMOLOGY INC
Grant note: NEI: 1R21EY034198, 1R01EY 037135 NSF: 2227373
Funded by NEI 1R21EY034198, NSF 2227373, NEI 1R01EY 037135.
Language: English
Date published: 04/01/2025
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Research Administration; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Dental Research; Chemical and Biochemical Engineering; Ophthalmology and Visual Sciences
Record Identifier: 9984808538502771

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