Disease-Corrected Hepatocyte-Like Cells from Familial Hypercholesterolemia-Induced Pluripotent Stem Cells
Disease-Corrected Hepatocyte-Like Cells from Familial Hypercholesterolemia-Induced Pluripotent Stem Cells
复制标题
DOI：
10.1007/s12033-012-9635-3
复制
发表时间：
2013-07-01
影响因子：
2.6
通讯作者：
Baharvand, Hossein
中科院分区：
文献类型：
--
作者：
Fattahi, Faranak;Asgari, Samira;Baharvand, Hossein
从单个患者中产生人类诱导多能干细胞(HiPSCs)为疾病建模、药物发现和细胞替代治疗提供了独特的工具。患者特有的多能干细胞可以在体外扩增，因此适合进行基因操作。到目前为止，已经使用患者特有的HiPSCs对几种遗传性肝病进行了建模。在这里，我们提出了从低密度脂蛋白受体(LDLR)基因纯合突变的家族性高胆固醇血症(FH)患者的HiPSCs中产生校正的肝细胞样细胞(HLCs)。我们从一位FH患者身上获得了HiPSCs，该突变基因编码了一个截短的非功能受体。为了将正常的LDLR传递给缺陷细胞，我们使用携带正常受体ORF的质粒载体对HiPSCs进行遗传转化。转化的细胞扩增后定向向HLCs方向分化。未分化的缺陷HiPSCs和由缺陷的HiPSC分化而来的HLCs不具有摄取标记的低密度脂蛋白(LDL)颗粒的能力。分化后的hPSCs具有摄取低密度脂蛋白的能力，具有纠正疾病表型和表达肝细胞特异性标志物的能力。吲哚青绿(ICG)摄取实验、PAS染色、诱导细胞色素P450活性和油红染色也证实了分化细胞的功能。这些数据表明，HiPSC技术可以用于产生疾病校正的、患者特异性的HLC，在未来的疾病建模和药物发现以及细胞治疗应用中具有潜在的价值。
The generation of human induced pluripotent stem cells (hiPSCs) from an individual patient provides a unique tool for disease modeling, drug discovery, and cell replacement therapies. Patient-specific pluripotent stem cells can be expanded in vitro and are thus suitable for genetic manipulations. To date, several genetic liver disorders have been modeled using patient-specific hiPSCs. Here, we present the generation of corrected hepatocyte-like cells (HLCs) from hiPSCs of a familial hypercholesterolemia (FH) patient with a homozygous mutation in the low-density lipoprotein receptor (LDLR) gene. We generated hiPSCs from a patient with FH with the mutated gene encoding a truncated non-functional receptor. In order to deliver normal LDLR to the defective cells, we used a plasmid vector carrying the normal receptor ORF to genetically transform the hiPSCs. The transformed cells were expanded and directed toward HLCs. Undifferentiated defective hiPSCs and HLCs differentiated from the defective hiPSCs did not have the ability to uptake labeled low-density lipoprotein (LDL) particles. The differentiated transformed hiPSCs showed LDL-uptake ability and the correction of disease phenotype as well as expressions of hepatocyte-specific markers. The functionality of differentiated cells was also confirmed by indo-cyanine green (ICG) uptake assay, PAS staining, inducible cyp450 activity, and oil red staining. These data suggest that hiPSC technology can be used for generation of disease-corrected, patient-specific HLCs with potential value for disease modeling and drug discovery as well as cell therapy applications in future.