Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform

碩士 === 國立陽明大學 === 藥理學研究所 === 105 === Leber’s hereditary optic neuropathy (LHON) is a maternally inherited mitochondrial disease caused by homoplasmic point mutations in Complex I subunit genes of mitochondrial DNA. LHON is characterized by incomplete penetrance, as only part of Complex I mutation ca...

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Main Authors: You-Ren Wu, 吳祐任
Other Authors: Shih-Hwa Chiou
Format: Others
Language:en_US
Published: 2017
Online Access:http://ndltd.ncl.edu.tw/handle/8z523p
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spelling ndltd-TW-105YM0055500252019-05-15T23:39:47Z http://ndltd.ncl.edu.tw/handle/8z523p Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform 利用誘導型多能幹細胞分化視網膜神經節細胞平台-探討利伯氏遺傳神經病變之致病機轉 You-Ren Wu 吳祐任 碩士 國立陽明大學 藥理學研究所 105 Leber’s hereditary optic neuropathy (LHON) is a maternally inherited mitochondrial disease caused by homoplasmic point mutations in Complex I subunit genes of mitochondrial DNA. LHON is characterized by incomplete penetrance, as only part of Complex I mutation carriers become affected. Therefore, the mitochondrial DNA mutation is necessary, but not sufficient to cause optic neuropathy. Multiple factors were considered for explaining its variable penetrance, such as environmental triggers and genetic modifying factors. However, the mechanism of low penetrance of Complex I mutation is still largely unclear. LHON patient-specific induced pluripotent stem cells (iPSCs) from affected patients and carriers provide a powerful in vitro model for evaluating the pathological phenotypes of the disease. First, we detected an identical homoplasmic 11778 G>A mutation in all individuals after RGCs differentiation to verify mitochondrial DNA mutation load would not affect by reprogramming and differentiation. To characterize the pathological phenotypes of LHON iPSC-RGCs, we compared the neurite outgrowth and mitochondrial function between control, affected and carriers. The common compensatory strategy to mitochondrial dysfunction observed in LHON iPSC-RGCs, a globally activated mitochondrial biogenesis associated with increased mitochondrial DNA content. The neurites morphology indicated affected individuals were defective in RGC neurogenesis and maintenance. We measured the mitochondrial Complex I enzyme activity and oxygen consumption, and showed that carriers have a significantly higher Complex I activity and affected have lower maximal respiratory capacity as compared to control. Furthermore, the whole genome profile and LHON-related pathways would be analyzed by microarray analysis in LHON carriers and patients. This study will provide a novel finding for the pathological mechanism of LHON. Shih-Hwa Chiou 邱士華 2017 學位論文 ; thesis 55 en_US
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description 碩士 === 國立陽明大學 === 藥理學研究所 === 105 === Leber’s hereditary optic neuropathy (LHON) is a maternally inherited mitochondrial disease caused by homoplasmic point mutations in Complex I subunit genes of mitochondrial DNA. LHON is characterized by incomplete penetrance, as only part of Complex I mutation carriers become affected. Therefore, the mitochondrial DNA mutation is necessary, but not sufficient to cause optic neuropathy. Multiple factors were considered for explaining its variable penetrance, such as environmental triggers and genetic modifying factors. However, the mechanism of low penetrance of Complex I mutation is still largely unclear. LHON patient-specific induced pluripotent stem cells (iPSCs) from affected patients and carriers provide a powerful in vitro model for evaluating the pathological phenotypes of the disease. First, we detected an identical homoplasmic 11778 G>A mutation in all individuals after RGCs differentiation to verify mitochondrial DNA mutation load would not affect by reprogramming and differentiation. To characterize the pathological phenotypes of LHON iPSC-RGCs, we compared the neurite outgrowth and mitochondrial function between control, affected and carriers. The common compensatory strategy to mitochondrial dysfunction observed in LHON iPSC-RGCs, a globally activated mitochondrial biogenesis associated with increased mitochondrial DNA content. The neurites morphology indicated affected individuals were defective in RGC neurogenesis and maintenance. We measured the mitochondrial Complex I enzyme activity and oxygen consumption, and showed that carriers have a significantly higher Complex I activity and affected have lower maximal respiratory capacity as compared to control. Furthermore, the whole genome profile and LHON-related pathways would be analyzed by microarray analysis in LHON carriers and patients. This study will provide a novel finding for the pathological mechanism of LHON.
author2 Shih-Hwa Chiou
author_facet Shih-Hwa Chiou
You-Ren Wu
吳祐任
author You-Ren Wu
吳祐任
spellingShingle You-Ren Wu
吳祐任
Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
author_sort You-Ren Wu
title Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
title_short Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
title_full Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
title_fullStr Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
title_full_unstemmed Pathogenic Investigation of Leber’s Hereditary Optic Neuropathy: Using Patient-Specific iPSCs-derived Retinal Ganglion Cells as a Diseased Platform
title_sort pathogenic investigation of leber’s hereditary optic neuropathy: using patient-specific ipscs-derived retinal ganglion cells as a diseased platform
publishDate 2017
url http://ndltd.ncl.edu.tw/handle/8z523p
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