HCN2 and TBX3 Reprogram Human-Induced Pluripotent Stem Cells-Derived Cardiomyocytes into Pacemaker-Like Cells
HCN2 and TBX3 Reprogram Human-Induced Pluripotent Stem Cells-Derived Cardiomyocytes into Pacemaker-Like Cells
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HCN2 和 TBX3 将人类诱导的多能干细胞来源的心肌细胞重新编程为起搏器样细胞
DOI：
10.1089/dna.2019.5135
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发表时间：
2020-01-09
影响因子：
3.1
通讯作者：
Huang, Congxin
中科院分区：
文献类型：
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作者：
Zhao, Hongyi;Wang, Fengyuan;Huang, Congxin
Tbx3将心肌细胞重编程为具有窦房结表型的细胞，但未检测到特异性滑链电流(IF)。我们探讨了过表达TBX3或与HCN2联合使用是否可以将人类诱导的多能干细胞来源的心肌细胞(hiPSC-CMS)重新编程为起搏器样细胞。本研究将TBX3和/或HCN_2基因导入HIPSC-CMS。表达分析表明，过表达TBX3导致工作心肌细胞降低到起搏细胞的表达谱。逆转录-定量聚合酶链式反应(RT-qPCR)和电生理分析显示，连接蛋白亚基(CX40、Cx43)、钠电流(SCN5A、Ina)、内向整流钾通道(Kir2.1、IK1)的表达减少，连接蛋白亚基(CX30.2、CX45)的表达增加。未检测到IF。Ik1的减少导致更多的去极化的最大舒张期电位和IF的表达(由HCN2产生)，它们协同作用产生自发的舒张期去极化，这是起搏细胞最典型的特征。总之，过表达TBX3和HCN2可以将HiPSC-CMS重新编程为起搏器样细胞。实现舒张期去极化形成的能力为构建生物起搏器提供了一种新的策略。
TBX3 reprograms cardiac myocytes into cells that possess sinoatrial node phenotype, but no specific funny current (If) was detected. We explore whether overexpression of TBX3 alone or combined with HCN2 can reprogram human-induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) into pacemaker-like cells. HiPSC-CMs were transfected with TBX3 and/or HCN2 in this study. Expression analysis showed that overexpression of TBX3 induces a reduced reduction expression profile of working cardiomyocytes into that of pacemaker cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and electrophysiological analyses showed a reduced expression of connexins subunits (CX40, CX43), the sodium current (SCN5A, INa), the inward rectified potassium channels (Kir2.1, IK1), and an increased expression of connexins subunits (CX30.2, CX45). No If was detected. The reduction of IK1 resulted in a more depolarized maximum diastolic potential together with an expression of If (generated by HCN2), which they work in synergy to generate spontaneous diastolic depolarization that was the most typical characteristic of pacemaker cells. In conclusion, overexpression of TBX3 and HCN2 could reprogram hiPSC-CMs into pacemaker-like cells. The ability to enable diastolic depolarization formation provides a new strategy for the construction of a biological pacemaker.