Repression of Zeb1 and hypoxia cause sequential mesenchymal-to-epithelial transition and induction of aid, Oct4, and Dnmt1, leading to immortalization and multipotential reprogramming of fibroblasts in spheres.
Repression of Zeb1 and hypoxia cause sequential mesenchymal-to-epithelial transition and induction of aid, Oct4, and Dnmt1, leading to immortalization and multipotential reprogramming of fibroblasts in spheres.
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DOI：
10.1002/stem.1382
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发表时间：
2013-07
期刊：
影响因子：
5.2
通讯作者：
Dean, Douglas C.
中科院分区：
文献类型：
--
作者：
Liu, Yongqing;Mukhopadhyay, Partha;Pisano, M. Michele;Lu, Xiaoqin;Huang, Li;Lu, Qingxian;Dean, Douglas C.
在这里，我们证明球体形成触发小鼠成纤维细胞的永生化和稳定重编程。球体中的细胞接触信号传导导致 EMT 转录因子 Zeb1 下调，从而导致间质到上皮的快速转变。并且，球体内的缺氧与 Zeb1 抑制作用的丧失协同作用，导致 Hif1a 的超诱导，进而导致 DNA 去甲基化酶 Aid/Aicda 的诱导、Oct4 启动子/增强子的去甲基化和多能性。当细胞从球体的缺氧环境中取出并置于单层培养物中时，Oct4 和 Nanog 的表达会减少，但细胞保留多潜能能力，表现出稳定的重编程和类似于成体干细胞的基因表达模式。 Oct4 已被证明可以在间充质干细胞中诱导 Dnmt1，我们将 Oct4 和 Dnmt1 与细胞周期抑制性细胞周期蛋白依赖性激酶抑制剂和 Arf 的沉默以及重编程成纤维细胞的永生化联系起来。球体形成代表了一种新颖且快速的方案，用于将成纤维细胞永生化和稳定重编程为多能性，不需要干细胞因子或谱系特异性转录因子的外源表达。
Here, we demonstrate that sphere formation triggers immortalization and stable reprogramming of mouse fibroblasts. Cell contact signaling in spheres causes downregulation of the EMT transcription factor Zeb1 leading to rapid mesenchymal-to-epithelial transition. And, hypoxia within spheres together with loss of Zeb1 repression synergize to cause superinduction of Hif1a, which in turn leads to induction of the DNA demethylase Aid/Aicda, demethylation of the Oct4 promoter/enhancer and multipotency. Oct4 and Nanog expression diminish when cells are removed from the hypoxic environment of spheres and placed in monolayer culture, but the cells retain multipotential capacity, demonstrating stable reprogramming and a gene expression pattern resembling adult stem cells. Oct4 has been shown to induce Dnmt1 in mesenchymal stem cells, and we link Oct4 and Dnmt1 to silencing of cell cycle inhibitory cyclin dependent kinase inhibitors and Arf, and immortalization of the reprogrammed fibroblasts. Sphere formation then represents a novel and rapid protocol for immortalization and stable reprogramming of fibroblasts to multipotency that does not require exogenous expression of a stem cell factor or a lineage-specifying transcription factor.
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影响因子：
64.5
作者：
Bhutani N;Burns DM;Blau HM
通讯作者：
Blau HM
影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
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Jaenisch, Rudolf
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11.2
作者：
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23.9
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Liu Y;Clem B;Zuba-Surma EK;El-Naggar S;Telang S;Jenson AB;Wang Y;Shao H;Ratajczak MZ;Chesney J;Dean DC
通讯作者：
Dean DC
影响因子：
64.5
作者：
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