Investigative Ophthalmology & Visual Science Cover Image for Volume 65, Issue 7
June 2024
Volume 65, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2024
Proteomics analysis of the regulation of proteins involved in steroid hormone signaling in immortalized human retinal pigment epithelial cells under oxidative stress
Author Affiliations & Notes
  • R Duncan
    Ophthalmology, University of Missouri-Kansas City, Kansas City, Missouri, United States
  • Conner Hall
    Ophthalmology, University of Missouri-Kansas City, Kansas City, Missouri, United States
  • Peter Koulen
    Ophthalmology, University of Missouri-Kansas City, Kansas City, Missouri, United States
  • Footnotes
    Commercial Relationships   R Duncan None; Conner Hall None; Peter Koulen None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2024, Vol.65, 6732. doi:
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      R Duncan, Conner Hall, Peter Koulen; Proteomics analysis of the regulation of proteins involved in steroid hormone signaling in immortalized human retinal pigment epithelial cells under oxidative stress. Invest. Ophthalmol. Vis. Sci. 2024;65(7):6732.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Oxidative stress can cause retinal pigment epithelium (RPE) dysfunction in age-related macular degeneration (AMD), but details on how RPE cells adapt to oxidative stress are incomplete. The current study was conducted to measure differentially expressed proteins that play a role in steroid hormone signaling in human telomerase reverse transcriptase (hTERT)-immortalized RPE cell line after oxidant exposure.

Methods : To induce oxidative stress, hTERT RPE-1 cells (ATCC# CRL-4000) were exposed to 100μM tBHP for 24 hours and were then processed for tandem mass tag (TMT) labeling, liquid chromatography (LC), LC-mass spectrometry and TMT quantitation. Data collection, reconciliation, organization, and Gene Ontology/functional category analyses were performed to identify differentially expressed proteins involved in steroid hormone signaling.

Results : analyses were performed to identify differentially expressed proteins involved in steroid hormone signaling.
Results: hTERT-RPE-1 cell exposure to tBHP led to a change in the expression of proteins found in the GO biological process categories ‘cellular response to glucocorticoid stimulus’ (GO:0071385), ‘response to glucocorticoid’ (GO:0051384), ‘regulation of androgen receptor signaling pathway’ (GO:0060765), and ‘intracellular steroid hormone receptor signaling pathway’ (GO:0030518). tBHP exposure led to the upregulation of 13 proteins involved in steroid hormone signaling by between 8.9- and 27.5-fold and decreased the expression of 11 such proteins by between 6.1- and 22.7-fold. Most of the upregulated proteins were protein chaperones/heat shock proteins while most downregulated proteins were involved in steroid hormone receptor-mediated gene expression.

Conclusions : Conditions of oxidative stress altered the expression levels of proteins that control steroid hormone signaling networks. We identified proteins that may play a significant adaptive and/or protective role in RPE cells and their susceptibility to oxidative stress in AMD.

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

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