Aberrant transcriptional networks in step-wise neurogenesis of paroxysmal kinesigenic dyskinesia-induced pluripotent stem cells.
Aberrant transcriptional networks in step-wise neurogenesis of paroxysmal kinesigenic dyskinesia-induced pluripotent stem cells.
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阵发性运动诱发性运动障碍诱导的多能干细胞逐步神经发生中的异常转录网络
DOI：
10.18632/oncotarget.10680
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发表时间：
2016-08-16
期刊：
影响因子：
--
通讯作者：
Jin Y
中科院分区：
文献类型：
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作者：
Li C;Ma Y;Zhang K;Gu J;Tang F;Chen S;Cao L;Li S;Jin Y
阵发性运动诱发性运动障碍（PKD）是一种常染色体显性遗传的发作性运动障碍，临床表现具有明显的变异性。富含脯氨酸的跨膜蛋白2（PRRT 2）已被确定为PKD的致病基因，但PKD发病的分子机制仍是一个谜。PKD疾病的表型和转录模式需要进一步阐明。在这里，我们报告了来自两个家族性PKD患者的iPSC系的产生和神经分化，这两个家族性PKD患者分别具有c.487C>T（p.Gln163X）和c.573dupT（p.Gly192Trpfs *8）PRRT 2突变。值得注意的是，通过SMAD信号传导的双重抑制的逐步神经分化方法观察到PKD-iPSC的神经转化效率比对照iPSC低得多。此外，我们首次显示了PRRT 2在整个人脑中的高表达水平以及PRRT 2在其他人体组织中的表达模式。为了从分子水平深入了解疾病的发展，我们在神经诱导的四个不同阶段对PKD细胞进行了全球基因表达谱分析，并确定了改变的基因表达模式，与对照组相比，这些基因表达模式特别反映了失调的神经转录组特征和中胚层发育的分化趋势。此外，功能和信号传导途径分析表明PKD-iPSC和对照iPSC之间的细胞命运决定显著不同。总之，PKD特异性体外模型的建立和PKD细胞中转录组特征的说明将有助于我们更好地理解神经转换的缺陷以及进一步研究PKD疾病的发病机制。
Paroxysmal kinesigenic dyskinesia (PKD) is an episodic movement disorder with autosomal-dominant inheritance and marked variability in clinical manifestations. Proline-rich transmembrane protein 2 (PRRT2) has been identified as a causative gene of PKD, but the molecular mechanism underlying the pathogenesis of PKD still remains a mystery. The phenotypes and transcriptional patterns of the PKD disease need further clarification. Here, we report the generation and neural differentiation of iPSC lines from two familial PKD patients with c.487C>T (p. Gln163X) and c.573dupT (p. Gly192Trpfs*8) PRRT2 mutations, respectively. Notably, an extremely lower efficiency in neural conversion from PKD-iPSCs than control-iPSCs is observed by a step-wise neural differentiation method of dual inhibition of SMAD signaling. Moreover, we show the high expression level of PRRT2 throughout the human brain and the expression pattern of PRRT2 in other human tissues for the first time. To gain molecular insight into the development of the disease, we conduct global gene expression profiling of PKD cells at four different stages of neural induction and identify altered gene expression patterns, which peculiarly reflect dysregulated neural transcriptome signatures and a differentiation tendency to mesodermal development, in comparison to control-iPSCs. Additionally, functional and signaling pathway analyses indicate significantly different cell fate determination between PKD-iPSCs and control-iPSCs. Together, the establishment of PKD-specific in vitro models and the illustration of transcriptome features in PKD cells would certainly help us with better understanding of the defects in neural conversion as well as further investigations in the pathogenesis of the PKD disease.