TY  - BOOK
AU  - Berinstein, Daniel M
TI  - Repressed SIRT1/PGC-1alpha pathway and mitochondrial disintegration in iPSC-derived RPE disease model of age-related macular degeneration
SN  - 1479-5876
PY  - 2016///
KW  - *Induced Pluripotent Stem Cells/pa [Pathology]
KW  - *Macular Degeneration/me [Metabolism]
KW  - *Macular Degeneration/pa [Pathology]
KW  - *Mitochondria/me [Metabolism]
KW  - *Models, Biological
KW  - *Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/me [Metabolism]
KW  - *Retinal Pigment Epithelium/pa [Pathology]
KW  - *Sirtuin 1/me [Metabolism]
KW  - Aged
KW  - Aged, 80 and over
KW  - Cell Differentiation
KW  - Female
KW  - Humans
KW  - Induced Pluripotent Stem Cells/me [Metabolism]
KW  - Induced Pluripotent Stem Cells/ul [Ultrastructure]
KW  - Male
KW  - Middle Aged
KW  - Oxidative Stress
KW  - Phagocytosis
KW  - Phenotype
KW  - Reactive Oxygen Species/me [Metabolism]
KW  - Retinal Pigment Epithelium/ul [Ultrastructure]
KW  - Signal Transduction
KW  - Skin/pa [Pathology]
KW  - Superoxide Dismutase/me [Metabolism]
KW  - MedStar Washington Hospital Center
KW  - Ophthalmology
KW  - Journal Article
N1  - Available online through MWHC library: 2003 - present
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; CONCLUSIONS: Our studies suggest SIRT1/PGC-1alpha 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; 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-1alpha), 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
UR  - https://dx.doi.org/10.1186/s12967-016-1101-8
ER  -