Repressed SIRT1/PGC-1α pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration

Journal of Translational Medicine

Research

Shuk Kei Cheng¹  Hong Cao¹  Yi Chu¹  Nady Golestaneh²  Daniel M. Berinstein³  Eugenia Poliakov⁴ 

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

关键词: AMD;    RPE;    Oxidative stress;    Mitochondria;    Cell viability;    ROS;    PGC-1α;    SIRT1;   

DOI  :  10.1186/s12967-016-1101-8

 received in 2016-11-28, accepted in 2016-11-29,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundStudy 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.MethodsGeneration 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.ResultsThe 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.ConclusionsOur 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.

【 授权许可】

CC BY   
© The Author(s) 2016