MicroRNA-302 increases reprogramming efficiency via repression of NR2F2.
MicroRNA-302 increases reprogramming efficiency via repression of NR2F2.
复制标题
DOI：
10.1002/stem.1278
复制
发表时间：
2013-02
期刊：
影响因子：
5.2
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
--
作者：
Hu, Shijun;Wilson, Kitchener D.;Ghosh, Zhumur;Han, Leng;Wang, Yongming;Lan, Feng;Ransohoff, Katherine J.;Burridge, Paul;Wu, Joseph C.
MicroRNA (miRNA) 通过翻译抑制和 RNA 衰减已成为基因表达的关键调节因子，并参与细胞分化、增殖、血管生成和细胞凋亡的调节。在这项研究中，我们使用 miRNA 和 mRNA 微阵列的全局生物信息学分析来预测人胚胎干细胞 (hESC) 和诱导多能干细胞 (iPSC) 中新的 miRNA-mRNA 相互作用。特别是，我们证明了 miR-302 簇与两个转录因子 NR2F2 和 OCT4 之间的调节反馈环。我们的数据显示 miR-302 和 OCT4 在多能细胞中高表达，而 NR2F2 仅在分化细胞中表达。靶标分析预测 NR2F2 是 miR-302 的直接靶标，我们通过报告荧光素酶测定和实时 PCR 实验证实了这一点。我们还证明 NR2F2 直接抑制 OCT4 启动子的活性，从而减少 OCT4 和 miR-302 之间的正反馈环。重要的是，与使用四种因子（“KMOS”）相比，当我们使用四种因子（KLF4、C-MYC、OCT4 和 SOX2）加 miR-302（这种重编程混合物以下称为“KMOS3”）将人类脂肪源性干细胞（hASC）重编程为 iPSC 时，获得了更高的重编程效率。此外，NR2F2 的 shRNA 敲低也通过增强重编程效率来模拟 miR-302 的过度表达。有趣的是，我们无法单独从 miR-302a/b/c/d 生成 iPSC，这与之前报道的 miR-302 本身可以重新编程人类皮肤癌细胞和人类毛囊细胞的出版物形成鲜明对比。总而言之，这些发现表明 miR-302 抑制 NR2F2 并通过 OCT4 的间接正向调节促进多能性。这种反馈回路代表了理解和诱导体细胞多能性的重要新机制。
MicroRNAs (miRNAs) have emerged as critical regulators of gene expression through translational inhibition and RNA decay, and have been implicated in the regulation of cellular differentiation, proliferation, angiogenesis, and apoptosis. In this study, we use global bioinformatics analysis of miRNA and mRNA microarrays to predict novel miRNA-mRNA interactions in human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). In particular, we demonstrate a regulatory feedback loop between the miR-302 cluster and two transcription factors, NR2F2 and OCT4. Our data show high expression of miR-302 and OCT4 in pluripotent cells, while NR2F2 is expressed exclusively in differentiated cells. Target analysis predicts that NR2F2 is a direct target of the miR-302, which we experimentally confirm by reporter luciferase assays and real-time PCR. We also demonstrate that NR2F2 directly inhibits the activity of the OCT4 promoter and thus diminishes the positive feedback loop between OCT4 and the miR-302. Importantly, higher reprogramming efficiencies were obtained when we reprogrammed human adipose-derived stem cells (hASCs) into iPSCs using four factors (KLF4, C-MYC, OCT4, and SOX2) plus miR-302 (this reprogramming cocktail is hereafter referred to as “KMOS3”) when compared to using four factors (“KMOS”). Furthermore, shRNA knockdown of NR2F2 mimics the over-expression of miR-302 by also enhancing reprogramming efficiency. Interestingly, we were unable to generate iPSCs from miR-302a/b/c/d alone, which is in contrast to previous publications that have reported that miR-302 by itself can reprogram human skin cancer cells and human hair follicle cells. Taken together, these findings demonstrate that miR-302 inhibits NR2F2 and promotes pluripotency through indirect positive regulation of OCT4. This feedback loop represents an important new mechanism for understanding and inducing pluripotency in somatic cells.
登录
查看更多内容
影响因子：
37.8
作者：
Hu S;Huang M;Li Z;Jia F;Ghosh Z;Lijkwan MA;Fasanaro P;Sun N;Wang X;Martelli F;Robbins RC;Wu JC
通讯作者：
Wu JC
小鼠子宫胚胎着床过程中MicroRNA的表达和调控
影响因子：
4.8
作者：
Hu, Shi-Jun;Ren, Gang;Yang, Zeng-Ming
通讯作者：
Yang, Zeng-Ming
影响因子：
64.5
作者：
Lewis, BP;Shih, IH;Burge, CB
通讯作者：
Burge, CB
影响因子：
4
作者：
Ambros, Victor
通讯作者：
Ambros, Victor
Mirbase：MicroRNA基因组学的工具。
影响因子：
14.9
作者：
Griffiths-Jones, Sam;Saini, Harpreet Kaur;van Dongen, Stijn;Enright, Anton J.
通讯作者：
Enright, Anton J.