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A synthetic DNA-binding inhibitor of SOX2 guides human induced pluripotent stem cells to differentiate into mesoderm
Nucleic Acids Research ( IF 16.6 ) Pub Date : 2017-07-31 , DOI: 10.1093/nar/gkx693
Junichi Taniguchi , Ganesh N. Pandian , Takuya Hidaka , Kaori Hashiya , Toshikazu Bando , Kyeong Kyu Kim , Hiroshi Sugiyama

Targeted differentiation of human induced pluripotent stem cells (hiPSCs) using only chemicals would have value-added clinical potential in the regeneration of complex cell types including cardiomyocytes. Despite the availability of several chemical inhibitors targeting proteins involved in signaling pathways, no bioactive synthetic DNA-binding inhibitors, targeting key cell fate-controlling genes such as SOX2, are yet available. Here, we demonstrate a novel DNA-based chemical approach to guide the differentiation of hiPSCs using pyrrole–imidazole polyamides (PIPs), which are sequence-selective DNA-binding synthetic molecules. Harnessing knowledge about key transcriptional changes during the induction of cardiomyocyte, we developed a DNA-binding inhibitor termed PIP-S2, targeting the 5′-CTTTGTT-3′ and demonstrated that inhibition of SOX2–DNA interaction by PIP-S2 triggers the mesoderm induction in hiPSCs. Genome-wide gene expression analyses revealed that PIP-S2 induced mesoderm by targeted alterations in SOX2-associated gene regulatory networks. Also, employment of PIP-S2 along with a Wnt/β-catenin inhibitor successfully generated spontaneously contracting cardiomyocytes, validating our concept that DNA-binding inhibitors could drive the directed differentiation of hiPSCs. Because PIPs can be fine-tuned to target specific DNA sequences, our DNA-based approach could be expanded to target and regulate key transcription factors specifically associated with desired cell types.

中文翻译:

合成的SOX2 DNA结合抑制剂可指导人类诱导的多能干细胞分化为中胚层

仅使用化学物质对人类诱导的多能干细胞(hiPSC)进行靶向分化,将在包括心肌细胞在内的复杂细胞类型的再生中具有增值的临床潜力。尽管有几种化学抑制剂可靶向信号通路中涉及的蛋白质,但还没有针对关键细胞命运控制基因(如SOX2)的生物活性合成DNA结合抑制剂。在这里,我们展示了一种新颖的基于DNA的化学方法,该方法可使用吡咯-咪唑聚酰胺(PIP)来指导hiPSC的分化,吡咯-咪唑聚酰胺(PIP)是具有序列选择性的DNA结合合成分子。利用关于心肌细胞诱导过程中关键转录变化的知识,我们开发了一种称为PIP-S2的DNA结合抑制剂,针对5'-CTTTGTT-3',并证明了PIP-S2对SOX2-DNA相互作用的抑制触发了hiPSC中的中胚层诱导。全基因组基因表达分析表明,PIP-S2通过SOX2相关基因调控网络中的定向改变诱导中胚层。同样,与Wnt /β-catenin抑制剂一起使用PIP-S2成功地产生了自发收缩的心肌细胞,证实了我们的观点,即DNA结合抑制剂可以驱动hiPSC的定向分化。由于可以对PIP进行微调以靶向特定的DNA序列,因此我们基于DNA的方法可以扩展为靶向和调节与所需细胞类型特异性相关的关键转录因子。
更新日期:2017-09-21
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