June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Multi-omics based metabolic profiling of the retinal pigment epithelium
Author Affiliations & Notes
  • Lisa Grönnert
    Roche Pharma Research and Early Development, Ophthalmology Discovery, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
    Faculty of Science, Universitat Zurich, Zurich, ZH, Switzerland
  • David Avila
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Juliane Siebourg-Polster
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Fabian Köchl
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Manuel Tzouros
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Megana Prasad
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Javier Gayan
    Roche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Peter Westenskow
    Roche Pharma Research and Early Development, Ophthalmology Discovery, Roche Innovation Center Basel, F.Hoffmann-La Roche Ltd, Basel, Basel-Stadt, Switzerland
  • Footnotes
    Commercial Relationships   Lisa Grönnert, F.Hoffmann-La Roche Ltd. (E); David Avila, F.Hoffmann-La Roche Ltd. (E); Juliane Siebourg-Polster, F.Hoffmann-La Roche Ltd. (E); Fabian Köchl, F.Hoffmann-La Roche Ltd. (E); Manuel Tzouros, F.Hoffmann-La Roche Ltd. (E); Megana Prasad, F.Hoffmann-La Roche Ltd. (E); Javier Gayan, F.Hoffmann-La Roche Ltd. (E); Peter Westenskow, F.Hoffmann-La Roche Ltd. (E)
  • Footnotes
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Investigative Ophthalmology & Visual Science June 2021, Vol.62, 2229. doi:
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      Lisa Grönnert, David Avila, Juliane Siebourg-Polster, Fabian Köchl, Manuel Tzouros, Megana Prasad, Javier Gayan, Peter Westenskow; Multi-omics based metabolic profiling of the retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 2021;62(8):2229.

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

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Abstract

Purpose : Photoreceptors rely on continuous nutrient delivery from the choriocapillaris via the retinal pigment epithelium (RPE). With advancing age, choriocapillaris atrophy and reduced choroidal blood flow create a hypoxic environment disturbing metabolism in the RPE that ceases metabolic support for the photoreceptors. Chronic ischemia can induce photoreceptor dysfunction and degeneration, hallmark features of age-related retinal diseases. Learning how hypoxia drives the metabolic stress response in RPE could give insights into hypoxia-induced retinal pathologies and identify strategies to prevent RPE, photoreceptor and vision loss. In this study, we perform proteomic, metabolomic and transcriptomic analyses to characterize the metabolic effects of hypoxia in RPE.

Methods : Primary human retinal pigment epithelial cells were cultured in normoxic (21% O2) or hypoxic (4% O2) conditions. After 48 hours, intracellular proteomic and metabolomic profiles were determined using quantitative proteomics and metabolomics (CE-TOF-MS and CE-QqQ-MS). RNA-Seq based transcriptomics was conducted after 4, 8, 12, 24, 48 and 72 hours of hypoxia, respectively. Differential expression and pathway analysis of hypoxia vs normoxia was performed per dataset and timepoint. 48 hour results for all experiments were then assessed jointly.

Results : Hypoxia induced a dependency on anaerobic glycolysis, consistent with increased glucose uptake and lactic acid production. TCA cycle metabolite levels indicate that in hypoxic RPE, reductive carboxylation is favored, whereas oxidative phosphorylation is suppressed. Despite a substantial decrease in ATP, hypoxic RPE maintained an energy status comparable to that of normoxic cells, presumably due to downregulation of anabolic pathways. Reduction in total glutathione and a decreased glutathione redox ratio indicate that hypoxia enhances oxidative stress.

Conclusions : Our study provides a comprehensive metabolic profile of hypoxic RPE that adds to the understanding of hypoxia-induced metabolic reprogramming as a driving force of age-related degenerative diseases in the retina. If confirmed by prospective studies, these findings can be used to develop novel strategies for modifying the metabolic stress response in hypoxic RPE and prevent vision loss.

This is a 2021 ARVO Annual Meeting abstract.

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