Mitochondrial gene replacement in human pluripotent stem cell-derived neural progenitors

被引：0
作者：
S Iyer
E Xiao
K Alsayegh
N Eroshenko
M J Riggs
J P Bennett
R R Rao
机构：
[1] Center for the Study of Biological Complexity,Department of Chemical and Life Science Engineering
[2] Life Sciences Program,Department of Human and Molecular Genetics
[3] Virginia Commonwealth University,Department of Neurology
[4] Virginia Commonwealth University,undefined
[5] Virginia Commonwealth University,undefined
[6] School of Engineering and Applied Sciences,undefined
[7] Harvard University,undefined
[8] Integrated Life Sciences Program,undefined
[9] Virginia Commonwealth University,undefined
[10] Parkinson's Disease Center,undefined
[11] Virginia Commonwealth University,undefined
[12] Virginia Commonwealth University,undefined
[13] Current address: Neuroimaging Core; Genes,undefined
[14] Cognition,undefined
[15] and Psychosis Program,undefined
[16] National Institute of Mental Health,undefined
[17] Bethesda,undefined
[18] MD,undefined
[19] USA.,undefined
来源：
Gene Therapy | 2012年 / 19卷
关键词：
neural progenitors; stem cells; mitochondrial DNA; TFAM; neurodegeneration;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Human pluripotent stem cell-derived neural progenitor (hNP) cells are an excellent resource for understanding early neural development and neurodegenerative disorders. Given that many neurodegenerative disorders can be correlated with defects in the mitochondrial genome, optimal utilization of hNP cells requires an ability to manipulate and monitor changes in the mitochondria. Here, we describe a novel approach that uses recombinant human mitochondrial transcription factor A (rhTFAM) protein to transfect and express a pathogenic mitochondrial genome (mtDNA) carrying the G11778A mutation associated with Leber's hereditary optic neuropathy (LHON) disease, into dideoxycytidine (ddC)-treated hNPs. Treatment with ddC reduced endogenous mtDNA and gene expression, without loss of hNP phenotypic markers. Entry of G11778A mtDNA complexed with the rhTFAM was observed in mitochondria of ddC-hNPs. Expression of the pathogenic RNA was confirmed by restriction enzyme analysis of the SfaN1-digested cDNA. On the basis of the expression of neuron-specific class III beta-tubulin, neuronal differentiation occurred. Our results show for the first time that pathogenic mtDNA can be introduced and expressed into hNPs without loss of phenotype or neuronal differentiation potential. This mitochondrial gene replacement technology allows for creation of in vitro stem cell-based models useful for understanding neuronal development and treatment of neurodegenerative disorders.
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页码：469 / 475
页数：6
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