Global gene expression profiles of mouse ESCs, disease-specific iPSCs, and gene-corrected iPSCs. Mus musculus

The use of patient-derived induced pluripotent stem (iPS) cells as treatment for genetic diseases entails genetic repair or transfer of genetic information as a prerequisite. We have chosen the murine model of tyrosinemia type 1 (fumarylacetoacetate hydrolase deficiency; FAH(-/-) mice) as a paradigm for hereditary metabolic liver disorders and evaluated fibroblast-derived FAH(-/-)-iPS cell lines as targets for gene correction. By aggregating FAH(-/-)-iPS cells with tetraploid embryos, we obtained FAH-/--iPS cell–derived mice, which exhibited the phenotype of the founding FAH(-/-)-mice. We then rescued the diseased phenotype by lentiviral transduction of FAH-cDNA and performed embryo aggregation with these gene-corrected FAHgc-iPS cells to obtain viable healthy mice. Our results demonstrate that iPS cell technology is a valid approach to establish mouse disease models directly from somatic cells bearing genetic defects. Furthermore, established iPS cell lines can be genetically manipulated without loss of pluripotency for treatment of genetic diseases. Overall design: 6 samples: 1 MEF, 1ESC, 4 iPSCs

以患者来源的诱导多能干细胞（induced pluripotent stem cells，iPS细胞）作为遗传疾病的治疗手段，需以基因修复或遗传信息转移为前提。本研究选取1型酪氨酸血症（延胡索酰乙酰乙酸水解酶缺陷；FAH(-/-)小鼠）的小鼠模型作为遗传性代谢性肝病的研究范式，并评估了成纤维细胞来源的FAH(-/-)-iPS细胞系作为基因校正靶点的可行性。通过将FAH(-/-)-iPS细胞与四倍体胚胎聚合，我们获得了FAH(-/-)-iPS细胞来源的小鼠，其表型与原始FAH(-/-)小鼠一致。随后我们通过慢病毒转导FAH互补DNA（cDNA）挽救了患病表型，并使用这些基因校正后的FAHgc-iPS细胞进行胚胎聚合，获得了存活的健康小鼠。本研究结果证实，iPS细胞技术可直接从携带遗传缺陷的体细胞构建小鼠疾病模型，是一种可靠的方法。此外，已建立的iPS细胞系可在不丧失多能性的前提下进行遗传操作，用于遗传疾病的治疗。实验整体设计：共6份样本：1份小鼠胚胎成纤维细胞（mouse embryonic fibroblasts，MEF）、1份胚胎干细胞（embryonic stem cells，ESC）以及4份iPS细胞系。