Disease-causing mitochondrial heteroplasmy segregated within induced pluripotent stem cell clones derived from a patient with MELAS.
Disease-causing mitochondrial heteroplasmy segregated within induced pluripotent stem cell clones derived from a patient with MELAS.
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DOI：
10.1002/stem.1389
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发表时间：
2013-07
期刊：
影响因子：
5.2
通讯作者：
Nelson, Timothy J.
中科院分区：
文献类型：
--
作者：
Folmes, Clifford D. L.;Martinez-Fernandez, Almudena;Perales-Clemente, Ester;Li, Xing;Mcdonald, Amber;Oglesbee, Devin;Hrstka, Sybil C.;Perez-Terzic, Carmen;Terzic, Andre;Nelson, Timothy J.
线粒体疾病根据突变型和野生型线粒体DNA(MtDNA)的混合物表现出病理表型，称为异质性。在此，我们检测了核重编程和克隆分离诱导的多能干细胞(IPSC)对线粒体异质性的影响。患有典型线粒体缺陷的患者来源的真皮成纤维细胞被诊断为MELAS，由于复合体I的ND5亚单位G13513A位置的异质性，患者来源的真皮成纤维细胞表现出线粒体功能障碍和氧化储备减少。与原始患者来源的成纤维细胞的细胞嵌合体一致，MELAS-IPSC克隆包含致病突变的类似范围的mtDNA异质性，其余mtDNA具有相同的轮廓。高异质性的IPSC克隆被用来证明延长干细胞传代足以清除突变的mtDNA，导致具有不同程度致病基因的等基因IPSC亚克隆。在IPSC克隆的比较分化中，与具有高异质性的等基因克隆相比，含有较低异质性的IPSC克隆具有更高的心源性产量。因此，患者来源的干细胞系中的mtDNA异质分离能够直接比较祖细胞和谱系受限的后代中的基因型/表型关系，并表明细胞命运的决定是作为mtDNA突变负荷的函数来调节的。这个基于核重新编程的新型模型系统引入了一种盘中病工具，用于检查生物工程组织中MELAS患者突变基因的影响，并引入了一种细胞探针，用于研究个别线粒体疾病的分子特征。
Mitochondrial diseases display pathological phenotypes according to the mixture of mutant versus wild-type mitochondrial DNA (mtDNA), known as heteroplasmy. We herein examined the impact of nuclear reprogramming and clonal isolation of induced pluripotent stem cells (iPSC) on mitochondrial heteroplasmy. Patient-derived dermal fibroblasts with a prototypical mitochondrial deficiency diagnosed as MELAS demonstrated mitochondrial dysfunction with reduced oxidative reserve due to heteroplasmy at position G13513A in the ND5 subunit of complex I. Bioengineered iPSC clones acquired pluripotency with multi-lineage differentiation capacity and demonstrated reduction in mitochondrial density and oxygen consumption distinguishing them from the somatic source. Consistent with the cellular mosaicism of the original patient-derived fibroblasts, the MELAS-iPSC clones contained a similar range of mtDNA heteroplasmy of the disease-causing mutation with identical profiles in the remaining mtDNA. High-heteroplasmy iPSC clones were used to demonstrate that extended stem cell passaging was sufficient to purge mutant mtDNA, resulting in isogenic iPSC subclones with various degrees of disease-causing genotypes. Upon comparative differentiation of iPSC clones, improved cardiogenic yield was associated with iPSC clones containing lower heteroplasmy compared to isogenic clones with high heteroplasmy. Thus, mtDNA heteroplasmic segregation within patient-derived stem cell lines enables direct comparison of genotype/phenotype relationships in progenitor cells and lineage-restricted progeny, and indicates that cell fate decisions are regulated as a function of mtDNA mutation load. The novel nuclear reprogramming-based model system introduces a disease-in-a-dish tool to examine the impact of mutant genotypes for MELAS patients in bioengineered tissues and a cellular probe for molecular features of individual mitochondrial diseases.
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影响因子：
56.9
作者：
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通讯作者：
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影响因子：
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29
作者：
Folmes CD;Nelson TJ;Martinez-Fernandez A;Arrell DK;Lindor JZ;Dzeja PP;Ikeda Y;Perez-Terzic C;Terzic A
通讯作者：
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