Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration

Deborah A. Ferrington; Mara C. Ebeling; Rebecca J. Kapphahn; Marcia R. Terluk; Cody R. Fisher; Jorge R. Polanco; Heidi Roehrich; Michaela M. Leary; Zhaohui Geng; James R. Dutton; Sandra R. Montezuma

doi:10.1016/j.redox.2017.05.015

Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration

Deborah A. Ferrington, Mara C. Ebeling, Rebecca J. Kapphahn, Marcia R. Terluk, Cody R. Fisher, Jorge R. Polanco, Heidi Roehrich, Michaela M. Leary, Zhaohui Geng, James R. Dutton, Sandra R. Montezuma

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Abstract

Age-related macular degeneration (AMD) is the leading cause of blindness among older adults. It has been suggested that mitochondrial defects in the retinal pigment epithelium (RPE) underlies AMD pathology. To test this idea, we developed primary cultures of RPE to ask whether RPE from donors with AMD differ in their metabolic profile compared with healthy age-matched donors. Analysis of gene expression, protein content, and RPE function showed that these cultured cells replicated many of the cardinal features of RPE in vivo. Using the Seahorse Extracellular Flux Analyzer to measure bioenergetics, we observed RPE from donors with AMD exhibited reduced mitochondrial and glycolytic function compared with healthy donors. RPE from AMD donors were also more resistant to oxidative inactivation of these two energy-producing pathways and were less susceptible to oxidation-induced cell death compared with cells from healthy donors. Investigation of the potential mechanism responsible for differences in bioenergetics and resistance to oxidative stress showed RPE from AMD donors had increased PGC1α protein as well as differential expression of multiple genes in response to an oxidative challenge. Based on our data, we propose that cultured RPE from donors phenotyped for the presence or absence of AMD provides an excellent model system for studying “AMD in a dish”. Our results are consistent with the ideas that (i) a bioenergetics crisis in the RPE contributes to AMD pathology, and (ii) the diseased environment in vivo causes changes in the cellular profile that are retained in vitro.

Original language	English (US)
Pages (from-to)	255-265
Number of pages	11
Journal	Redox Biology
Volume	13
DOIs	https://doi.org/10.1016/j.redox.2017.05.015
State	Published - Oct 2017

Bibliographical note

Funding Information:
This work was supported by the Elaine and Robert Larson Endowed Vision Research Chair (to DAF); National Institutes of Health/ National Institute of Aging (T32-AG29796 to MRT) and National Eye Institute (R01 EY026012 to DAF); Beckman Initiative for Macular Research (#1303); Foundation Fighting Blindness (TA-NMT-0613-0620-UMN); Regenerative Medicine Minnesota (2015-5337);an anonymous benefactor for Macular Degeneration Research; the Lindsay Family Foundation; Minnesota Lions Vision Foundation; an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology and Visual Neurosciences. None of the funding agencies had a role in study design, in the collection, analysis and interpretation of data, in writing the manuscript, or in the decision to submit the manuscript for publication.

Publisher Copyright:
© 2017 The Authors

Keywords

6 max) Age-related macular degeneration
Antioxidants
Glycolytic function
Mitochondrial function
Oxidative stress
Retinal pigment epithelium

Publisher link

10.1016/j.redox.2017.05.015

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466586

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Ferrington, D. A., Ebeling, M. C., Kapphahn, R. J., Terluk, M. R., Fisher, C. R., Polanco, J. R., Roehrich, H., Leary, M. M., Geng, Z., Dutton, J. R., & Montezuma, S. R. (2017). Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration. Redox Biology, 13, 255-265. https://doi.org/10.1016/j.redox.2017.05.015

Ferrington, DA, Ebeling, MC, Kapphahn, RJ, Terluk, MR, Fisher, CR, Polanco, JR, Roehrich, H, Leary, MM, Geng, Z , Dutton, JR & Montezuma, SR 2017, 'Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration', Redox Biology, vol. 13, pp. 255-265. https://doi.org/10.1016/j.redox.2017.05.015

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title = "Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration",

abstract = "Age-related macular degeneration (AMD) is the leading cause of blindness among older adults. It has been suggested that mitochondrial defects in the retinal pigment epithelium (RPE) underlies AMD pathology. To test this idea, we developed primary cultures of RPE to ask whether RPE from donors with AMD differ in their metabolic profile compared with healthy age-matched donors. Analysis of gene expression, protein content, and RPE function showed that these cultured cells replicated many of the cardinal features of RPE in vivo. Using the Seahorse Extracellular Flux Analyzer to measure bioenergetics, we observed RPE from donors with AMD exhibited reduced mitochondrial and glycolytic function compared with healthy donors. RPE from AMD donors were also more resistant to oxidative inactivation of these two energy-producing pathways and were less susceptible to oxidation-induced cell death compared with cells from healthy donors. Investigation of the potential mechanism responsible for differences in bioenergetics and resistance to oxidative stress showed RPE from AMD donors had increased PGC1α protein as well as differential expression of multiple genes in response to an oxidative challenge. Based on our data, we propose that cultured RPE from donors phenotyped for the presence or absence of AMD provides an excellent model system for studying “AMD in a dish”. Our results are consistent with the ideas that (i) a bioenergetics crisis in the RPE contributes to AMD pathology, and (ii) the diseased environment in vivo causes changes in the cellular profile that are retained in vitro.",

keywords = "6 max) Age-related macular degeneration, Antioxidants, Glycolytic function, Mitochondrial function, Oxidative stress, Retinal pigment epithelium",

author = "Ferrington, {Deborah A.} and Ebeling, {Mara C.} and Kapphahn, {Rebecca J.} and Terluk, {Marcia R.} and Fisher, {Cody R.} and Polanco, {Jorge R.} and Heidi Roehrich and Leary, {Michaela M.} and Zhaohui Geng and Dutton, {James R.} and Montezuma, {Sandra R.}",

note = "Funding Information: This work was supported by the Elaine and Robert Larson Endowed Vision Research Chair (to DAF); National Institutes of Health/ National Institute of Aging (T32-AG29796 to MRT) and National Eye Institute (R01 EY026012 to DAF); Beckman Initiative for Macular Research (#1303); Foundation Fighting Blindness (TA-NMT-0613-0620-UMN); Regenerative Medicine Minnesota (2015-5337);an anonymous benefactor for Macular Degeneration Research; the Lindsay Family Foundation; Minnesota Lions Vision Foundation; an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology and Visual Neurosciences. None of the funding agencies had a role in study design, in the collection, analysis and interpretation of data, in writing the manuscript, or in the decision to submit the manuscript for publication. Publisher Copyright: {\textcopyright} 2017 The Authors",

year = "2017",

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doi = "10.1016/j.redox.2017.05.015",

language = "English (US)",

volume = "13",

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journal = "Redox Biology",

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T1 - Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration

AU - Ferrington, Deborah A.

AU - Ebeling, Mara C.

AU - Kapphahn, Rebecca J.

AU - Terluk, Marcia R.

AU - Fisher, Cody R.

AU - Polanco, Jorge R.

AU - Roehrich, Heidi

AU - Leary, Michaela M.

AU - Geng, Zhaohui

AU - Dutton, James R.

AU - Montezuma, Sandra R.

N1 - Funding Information: This work was supported by the Elaine and Robert Larson Endowed Vision Research Chair (to DAF); National Institutes of Health/ National Institute of Aging (T32-AG29796 to MRT) and National Eye Institute (R01 EY026012 to DAF); Beckman Initiative for Macular Research (#1303); Foundation Fighting Blindness (TA-NMT-0613-0620-UMN); Regenerative Medicine Minnesota (2015-5337);an anonymous benefactor for Macular Degeneration Research; the Lindsay Family Foundation; Minnesota Lions Vision Foundation; an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology and Visual Neurosciences. None of the funding agencies had a role in study design, in the collection, analysis and interpretation of data, in writing the manuscript, or in the decision to submit the manuscript for publication. Publisher Copyright: © 2017 The Authors

PY - 2017/10

Y1 - 2017/10

N2 - Age-related macular degeneration (AMD) is the leading cause of blindness among older adults. It has been suggested that mitochondrial defects in the retinal pigment epithelium (RPE) underlies AMD pathology. To test this idea, we developed primary cultures of RPE to ask whether RPE from donors with AMD differ in their metabolic profile compared with healthy age-matched donors. Analysis of gene expression, protein content, and RPE function showed that these cultured cells replicated many of the cardinal features of RPE in vivo. Using the Seahorse Extracellular Flux Analyzer to measure bioenergetics, we observed RPE from donors with AMD exhibited reduced mitochondrial and glycolytic function compared with healthy donors. RPE from AMD donors were also more resistant to oxidative inactivation of these two energy-producing pathways and were less susceptible to oxidation-induced cell death compared with cells from healthy donors. Investigation of the potential mechanism responsible for differences in bioenergetics and resistance to oxidative stress showed RPE from AMD donors had increased PGC1α protein as well as differential expression of multiple genes in response to an oxidative challenge. Based on our data, we propose that cultured RPE from donors phenotyped for the presence or absence of AMD provides an excellent model system for studying “AMD in a dish”. Our results are consistent with the ideas that (i) a bioenergetics crisis in the RPE contributes to AMD pathology, and (ii) the diseased environment in vivo causes changes in the cellular profile that are retained in vitro.

AB - Age-related macular degeneration (AMD) is the leading cause of blindness among older adults. It has been suggested that mitochondrial defects in the retinal pigment epithelium (RPE) underlies AMD pathology. To test this idea, we developed primary cultures of RPE to ask whether RPE from donors with AMD differ in their metabolic profile compared with healthy age-matched donors. Analysis of gene expression, protein content, and RPE function showed that these cultured cells replicated many of the cardinal features of RPE in vivo. Using the Seahorse Extracellular Flux Analyzer to measure bioenergetics, we observed RPE from donors with AMD exhibited reduced mitochondrial and glycolytic function compared with healthy donors. RPE from AMD donors were also more resistant to oxidative inactivation of these two energy-producing pathways and were less susceptible to oxidation-induced cell death compared with cells from healthy donors. Investigation of the potential mechanism responsible for differences in bioenergetics and resistance to oxidative stress showed RPE from AMD donors had increased PGC1α protein as well as differential expression of multiple genes in response to an oxidative challenge. Based on our data, we propose that cultured RPE from donors phenotyped for the presence or absence of AMD provides an excellent model system for studying “AMD in a dish”. Our results are consistent with the ideas that (i) a bioenergetics crisis in the RPE contributes to AMD pathology, and (ii) the diseased environment in vivo causes changes in the cellular profile that are retained in vitro.

KW - 6 max) Age-related macular degeneration

KW - Antioxidants

KW - Glycolytic function

KW - Mitochondrial function

KW - Oxidative stress

KW - Retinal pigment epithelium

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Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration

Abstract

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