Enhanced generation of human induced pluripotent stem cells by ectopic expression of Connexin 45.
Enhanced generation of human induced pluripotent stem cells by ectopic expression of Connexin 45.
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通过异位表达 Connexin 45 增强人诱导多能干细胞的生成
DOI：
10.1038/s41598-017-00523-y
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发表时间：
2017-03-28
影响因子：
4.6
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Ke Q;Li L;Yao X;Lai X;Cai B;Chen H;Chen R;Zhai Z;Huang L;Li K;Hu A;Mao FF;Xiang AP;Tao L;Li W
体细胞可以通过特定转录因子的异位表达成功地重编程为多能干细胞。然而，提高效率和更好地理解重编程的分子机制仍然是需要的。在本研究中，刮擦加载/染料转移实验表明，人诱导多能干细胞（hiPSCs）含有部分由连接蛋白45 （CX45）贡献的功能性间隙连接。然后我们发现CX45在人胚胎干细胞（hESCs）和人真皮成纤维细胞（hDFs）衍生的hiPSCs中表达。然后我们发现CX45在重编程过程中显著上调。最重要的是，CX45的异位表达与Yamanaka因子（OCT4、SOX2、KLF4、cMYC - OSKM）一起显著提高了重编程效率，而内源性CX45的表达下调则显著阻断了细胞重编程，降低了重编程效率。我们的进一步研究表明，CX45过表达或敲低可调节细胞增殖速率，而细胞增殖速率与重编程效率有关。总之，我们的数据强调了CX45在重编程中的关键作用，可能会增加细胞分裂率，并导致iPSCs产生的加速动力学。
Somatic cells can be successfully reprogrammed into pluripotent stem cells by the ectopic expression of defined transcriptional factors. However, improved efficiency and better understanding the molecular mechanism underlying reprogramming are still required. In the present study, a scrape loading/dye transfer assay showed that human induced pluripotent stem cells (hiPSCs) contained functional gap junctions partially contributed by Connexin 45 (CX45). We then found CX45 was expressed in human embryonic stem cells (hESCs) and human dermal fibroblasts (hDFs) derived hiPSCs. Then we showed that CX45 was dramatically upregulated during the reprogramming process. Most importantly, the ectopic expression of CX45 significantly enhanced the reprogramming efficiency together with the Yamanaka factors (OCT4, SOX2, KLF4, cMYC - OSKM), whereas knockdown of endogenous CX45 expression significantly blocked cellular reprogramming and reduced the efficiency. Our further study demonstrated that CX45 overexpression or knockdown modulated the cell proliferation rate which was associated with the reprogramming efficiency. In conclusion, our data highlighted the critical role of CX45 in reprogramming and may increase the cell division rate and result in an accelerated kinetics of iPSCs production.