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Repressed SIRT1/PGC-1α pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration.
J Transl Med. 2016 12 20; 14(1):344.JT

Abstract

BACKGROUND

Study of age related macular degeneration (AMD) has been hampered by lack of human models that represent the complexity of the disease. Here we have developed a human in vitro disease model of AMD to investigate the underlying AMD disease mechanisms.

METHODS

Generation of iPSCs from retinal pigment epithelium (RPE) of AMD donors, age-matched normal donors, skin fibroblasts of a dry AMD patient, and differentiation of iPSCs into RPE (AMD RPE-iPSC-RPE, normal RPE-iPSC-RPE and AMD Skin-iPSC-RPE, respectively). Immunostaining, cell viability assay and reactive oxygen species (ROS) production under oxidative stress conditions, electron microscopy (EM) imaging, ATP production and glycogen concentration assays, quantitative real time PCR, western blot, karyotyping.

RESULTS

The AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE present functional impairment and exhibit distinct disease phenotypes compared to RPE-iPSC-RPE generated from normal donors (Normal RPE-iPSC-RPE). The AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE show increased susceptibility to oxidative stress and produced higher levels of reactive oxygen species (ROS) under stress in accordance with recent reports. The susceptibility to oxidative stress-induced cell death in AMD RPE-iPSC-RPE and Skin-iPSC-RPE was consistent with inability of the AMD RPE-iPSC-RPE and Skin-iPSC-RPE to increase SOD2 expression under oxidative stress. Phenotypic analysis revealed disintegrated mitochondria, accumulation of autophagosomes and lipid droplets in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE. Mitochondrial activity was significantly lower in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE compared to normal cells and glycogen concentration was significantly increased in the diseased cells. Furthermore, Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), a regulator of mitochondrial biogenesis and function was repressed, and lower expression levels of NAD-dependent deacetylase sirtuin1 (SIRT1) were found in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE as compared to normal RPE-iPSC-RPE.

CONCLUSIONS

Our studies suggest SIRT1/PGC-1α as underlying pathways contributing to AMD pathophysiology, and open new avenues for development of targeted drugs for treatment of this devastating neurodegenerative disease of the visual system.

Authors+Show Affiliations

Department of Ophthalmology, Georgetown University Medical Center, 3900 Reservoir Road NW, Medical-Dental Building, Room NE203, Washington, DC, 20057, USA. ncg8@georgetown.edu. Department of Neurology, Georgetown University Medical Center, Washington, DC, USA. ncg8@georgetown.edu. Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, Washington, DC, USA. ncg8@georgetown.edu.Department of Ophthalmology, Georgetown University Medical Center, 3900 Reservoir Road NW, Medical-Dental Building, Room NE203, Washington, DC, 20057, USA.Department of Ophthalmology, Georgetown University Medical Center, 3900 Reservoir Road NW, Medical-Dental Building, Room NE203, Washington, DC, 20057, USA.Department of Ophthalmology, Georgetown University Medical Center, 3900 Reservoir Road NW, Medical-Dental Building, Room NE203, Washington, DC, 20057, USA.Retinal Cell and Molecular Biology (LRCMB), National Eye Institute, National Institutes of Health, Bethesda, MD, USA.Retina Group of Washington, Chevy Chase, MD, 20815, USA.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

27998274

Citation

Golestaneh, Nady, et al. "Repressed SIRT1/PGC-1α Pathway and Mitochondrial Disintegration in iPSC-derived RPE Disease Model of Age-related Macular Degeneration." Journal of Translational Medicine, vol. 14, no. 1, 2016, p. 344.
Golestaneh N, Chu Y, Cheng SK, et al. Repressed SIRT1/PGC-1α pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration. J Transl Med. 2016;14(1):344.
Golestaneh, N., Chu, Y., Cheng, S. K., Cao, H., Poliakov, E., & Berinstein, D. M. (2016). Repressed SIRT1/PGC-1α pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration. Journal of Translational Medicine, 14(1), 344. https://doi.org/10.1186/s12967-016-1101-8
Golestaneh N, et al. Repressed SIRT1/PGC-1α Pathway and Mitochondrial Disintegration in iPSC-derived RPE Disease Model of Age-related Macular Degeneration. J Transl Med. 2016 12 20;14(1):344. PubMed PMID: 27998274.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Repressed SIRT1/PGC-1α pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration. AU - Golestaneh,Nady, AU - Chu,Yi, AU - Cheng,Shuk Kei, AU - Cao,Hong, AU - Poliakov,Eugenia, AU - Berinstein,Daniel M, Y1 - 2016/12/20/ PY - 2016/11/28/received PY - 2016/11/29/accepted PY - 2016/12/22/entrez PY - 2016/12/22/pubmed PY - 2017/10/21/medline KW - AMD KW - Cell viability KW - Mitochondria KW - Oxidative stress KW - PGC-1α KW - ROS KW - RPE KW - SIRT1 SP - 344 EP - 344 JF - Journal of translational medicine JO - J Transl Med VL - 14 IS - 1 N2 - BACKGROUND: Study of age related macular degeneration (AMD) has been hampered by lack of human models that represent the complexity of the disease. Here we have developed a human in vitro disease model of AMD to investigate the underlying AMD disease mechanisms. METHODS: Generation of iPSCs from retinal pigment epithelium (RPE) of AMD donors, age-matched normal donors, skin fibroblasts of a dry AMD patient, and differentiation of iPSCs into RPE (AMD RPE-iPSC-RPE, normal RPE-iPSC-RPE and AMD Skin-iPSC-RPE, respectively). Immunostaining, cell viability assay and reactive oxygen species (ROS) production under oxidative stress conditions, electron microscopy (EM) imaging, ATP production and glycogen concentration assays, quantitative real time PCR, western blot, karyotyping. RESULTS: The AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE present functional impairment and exhibit distinct disease phenotypes compared to RPE-iPSC-RPE generated from normal donors (Normal RPE-iPSC-RPE). The AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE show increased susceptibility to oxidative stress and produced higher levels of reactive oxygen species (ROS) under stress in accordance with recent reports. The susceptibility to oxidative stress-induced cell death in AMD RPE-iPSC-RPE and Skin-iPSC-RPE was consistent with inability of the AMD RPE-iPSC-RPE and Skin-iPSC-RPE to increase SOD2 expression under oxidative stress. Phenotypic analysis revealed disintegrated mitochondria, accumulation of autophagosomes and lipid droplets in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE. Mitochondrial activity was significantly lower in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE compared to normal cells and glycogen concentration was significantly increased in the diseased cells. Furthermore, Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), a regulator of mitochondrial biogenesis and function was repressed, and lower expression levels of NAD-dependent deacetylase sirtuin1 (SIRT1) were found in AMD RPE-iPSC-RPE and AMD Skin-iPSC-RPE as compared to normal RPE-iPSC-RPE. CONCLUSIONS: Our studies suggest SIRT1/PGC-1α as underlying pathways contributing to AMD pathophysiology, and open new avenues for development of targeted drugs for treatment of this devastating neurodegenerative disease of the visual system. SN - 1479-5876 UR - https://www.unboundmedicine.com/medline/citation/27998274/Repressed_SIRT1/PGC_1α_pathway_and_mitochondrial_disintegration_in_iPSC_derived_RPE_disease_model_of_age_related_macular_degeneration_ L2 - https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-016-1101-8 DB - PRIME DP - Unbound Medicine ER -