Urine-derived induced pluripotent stem cells as a modeling tool for paroxysmal kinesigenic dyskinesia.
Urine-derived induced pluripotent stem cells as a modeling tool for paroxysmal kinesigenic dyskinesia.
复制标题
尿源性诱导多能干细胞作为阵发性运动诱发性运动障碍的建模工具。
DOI：
10.1242/bio.013078
复制
发表时间：
2015-11-30
期刊：
影响因子：
2.4
通讯作者：
Wu ZY
中科院分区：
文献类型：
--
作者：
Zhang SZ;Li HF;Ma LX;Qian WJ;Wang ZF;Wu ZY
阵发性运动障碍（PKD）是一种常染色体显性遗传的单基因运动障碍。我们之前发现富含脯氨酸的跨膜蛋白2 （PRRT2）是PKD的致病基因。然而，到目前为止，PKD的发病机制仍不清楚。此外，还缺乏适用的建模工具来研究PKD的潜在机制。疾病特异性人类诱导多能干细胞（iPSCs）和定向细胞分化的结合为疾病建模提供了理想的平台。在这项研究中，我们从一名携带热点c.649dupC突变的PKD患者的肾上皮细胞（PKD- ipscs）中生成了两个iPSC系。这些细胞系碱性磷酸酶Nanog、Tra-1-80、Tra-1-60、SSEA-3和SSEA-4均呈阳性。在NOD/SCID小鼠体内注射PKD-iPSCs 2个月后获得具有外胚层、中胚层和内胚层三个胚层的畸胎瘤。与对照iPSCs相比，PKD-iPSCs中PRRT2 mRNA的表达降低。此外，PKD-iPSCs在体外具有向谷氨酸能、多巴胺能和运动神经元分化的潜能。电生理检查显示，快速激活和失活的钠通道以及电压门控钾通道的电流密度在PKD-iPSCs和对照iPSCs之间没有差异。因此，PKD- ipscs是研究PKD致病机制的可行建模工具。摘要：这是首例利用尿细胞诱导的多能干细胞研究阵发性运动障碍的病理机制的报道。
Paroxysmal kinesigenic dyskinesia (PKD) is a monogenic movement disorder with autosomal dominant inheritance. We previously identified the proline-rich transmembrane protein 2 (PRRT2) as a causative gene of PKD. However, the pathogenesis of PKD remains largely unknown so far. In addition, applicable modeling tools to investigate the underlying mechanisms of PKD are still lacking. The combination of disease-specific human induced pluripotent stem cells (iPSCs) and directed cell differentiation offers an ideal platform for disease modeling. In this study, we generated two iPSC lines from the renal epithelial cells of one PKD patient with the hotspot c.649dupC mutation (PKD-iPSCs). These cell lines were positive for alkaline phosphatase Nanog, Tra-1-80, Tra-1-60, SSEA-3 and SSEA-4. Teratomas with three blastoderms including ectoderm, mesoderm, and endoderm were obtained two months after injection of PKD-iPSCs into NOD/SCID mice. The expression of PRRT2 mRNA was decreased in PKD-iPSCs compared with that of the control iPSCs. Furthermore, PKD-iPSCs possessed the differentiation potential of functional glutamatergic, dopaminergic and motor neurons in vitro. Electrophysiological examinations revealed that the current densities of fast activated and deactivated sodium channels as well as voltage gated potassium channels were not different between the neurons from PKD-iPSCs and control iPSCs. Thus, PKD-iPSCs are a feasible modeling tool to investigate the pathogenic mechanisms of PKD. Summary: This is the first report of urinary cell-induced pluripotent stem cells being used as resources for investigation of the pathological mechanisms of paroxysmal kinesigenic dyskinesia.