A genome-wide RNAi screen reveals MAP kinase phosphatases as key ERK pathway regulators during embryonic stem cell differentiation.
A genome-wide RNAi screen reveals MAP kinase phosphatases as key ERK pathway regulators during embryonic stem cell differentiation.
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DOI：
10.1371/journal.pgen.1003112
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发表时间：
2012
期刊：
影响因子：
4.5
通讯作者：
Sharrocks AD
中科院分区：
文献类型：
--
作者：
Yang SH;Kalkan T;Morrisroe C;Smith A;Sharrocks AD
胚胎干细胞和诱导性多能干细胞代表了再生医学中潜在的重要治疗剂。复杂的相互关联的转录和信号网络控制这些细胞的命运，以维持多能性或分化。在这项研究中，我们集中在小鼠胚胎干细胞如何开始分化和失去多能性，特别是ERK MAP激酶和GSK 3信号通路在这一过程中发挥的作用。通过全基因组siRNA筛选，我们已经确定了400多个参与多能性丧失和促进分化的基因。这些基因在功能上与ERK和/或GSK 3通路相关，为研究这些通路在控制逃离多能基态中的作用提供了重要资源。更详细的分析确定了MAP激酶磷酸酶作为一个焦点的调节，并证明了这些酶在控制ERK激活动力学和随后确定早期胚胎干细胞的命运决定的重要作用。胚胎干细胞和诱导性多能干细胞代表了再生医学中潜在的重要治疗剂。对这些细胞类型的操纵可以让我们替换体内死亡或患病的细胞，从而为广泛的医学问题提供解决方案。然而，在我们进行这种操作之前，我们需要了解干细胞是如何连接的，以便我们能够以合乎逻辑的方式重新连接它们以产生所需的细胞类型。在这里，我们试图通过使用RNAi筛选来理解这种连接，在该筛选中，细胞的每个单独的组分被系统地去除，并确定对细胞命运的影响。我们已经确定了数百个基因，这些基因是有效丧失干细胞特征并因此转化为其他细胞类型所必需的。通过研究这些基因的一个子集，我们已经能够表明，许多人聚集在两个相关的负调节因子的关键途径之一，促进干细胞身份的丧失。这些负调节因子Dusps通常限制干细胞改变其功能的能力，从而转化为不同的细胞类型。
Embryonic stem cells and induced pluripotent stem cells represent potentially important therapeutic agents in regenerative medicine. Complex interlinked transcriptional and signaling networks control the fate of these cells towards maintenance of pluripotency or differentiation. In this study we have focused on how mouse embryonic stem cells begin to differentiate and lose pluripotency and, in particular, the role that the ERK MAP kinase and GSK3 signaling pathways play in this process. Through a genome-wide siRNA screen we have identified more than 400 genes involved in loss of pluripotency and promoting the onset of differentiation. These genes were functionally associated with the ERK and/or GSK3 pathways, providing an important resource for studying the roles of these pathways in controlling escape from the pluripotent ground state. More detailed analysis identified MAP kinase phosphatases as a focal point of regulation and demonstrated an important role for these enzymes in controlling ERK activation kinetics and subsequently determining early embryonic stem cell fate decisions. Embryonic stem cells and induced pluripotent stem cells represent potentially important therapeutic agents in regenerative medicine. Manipulation of these cell types could allow us to replace dead or diseased cells in our bodies and hence potentially provide a solution to a wide range of medical problems. However, before we can perform such manipulations, we need to understand how the stem cells are wired so that we are able to re-wire them in a logical way to produce the desired cell types. Here we have attempted to understand this wiring by using an RNAi screen in which each individual component of the cell is systematically removed and the consequences on cellular fate determined. We have identified hundreds of genes that are required for efficient loss of stem cell characteristics and hence conversion into other cell types. By studying a subset of these genes, we have been able to show that many converge on two related negative regulators of one of the key pathways that act to promote loss of stem cell identity. These negative regulators, Dusps, normally limit the ability of stem cells to change their function and hence be converted to different cell types.
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DOI：
10.1242/dev.037689
发表时间：
2010-11
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Dorey K;Amaya E
通讯作者：
Amaya E
影响因子：
64.5
作者：
Peng JC;Valouev A;Swigut T;Zhang J;Zhao Y;Sidow A;Wysocka J
通讯作者：
Wysocka J
影响因子：
34.7
作者：
Hur, Eun-Mi;Zhou, Feng-Quan
通讯作者：
Zhou, Feng-Quan
影响因子：
4.8
作者：
Asada, Sachie;Daitoku, Hiroaki;Fukamizu, Akiyoshi
通讯作者：
Fukamizu, Akiyoshi
影响因子：
10.5
作者：
Cole, Megan F.;Johnstone, Sarah E.;Young, Richard A.
通讯作者：
Young, Richard A.