Disabled-2 Is Required for Mesoderm Differentiation of Murine Embryonic Stem Cells
Disabled-2 Is Required for Mesoderm Differentiation of Murine Embryonic Stem Cells
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DOI：
10.1002/jcp.22200
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发表时间：
2010-10-01
影响因子：
5.6
通讯作者：
Tseng, Ching-Ping
中科院分区：
文献类型：
--
作者：
Huang, Chien-Ling;Cheng, Ju-Chien;Tseng, Ching-Ping
中胚层分化的控制涉及多种信号网络。先前的研究表明，残疾人-2（DAB 2）是一种多功能蛋白，参与生长因子信号传导和胚胎发育。在这项研究中，我们研究了小鼠胚胎干细胞（ESCs）体外中胚层分化过程中DAB 2的表达和功能。我们发现DAB 2在与OP 9基质细胞共培养的胚胎干细胞向中胚层分化时表达上调。胚胎干细胞诱导胚状体形成时，DAB 2也上调。DAB 2短发夹小干扰RNA（shDAB 2）的表达并没有改变胚胎干细胞的纯度。然而，shDAB 2破坏ESCs细胞-细胞粘附，影响胚状体和集落形成，随后阻碍ESCs的中胚层分化。免疫荧光染色显示，β-连环蛋白和斑珠蛋白细胞分布的紊乱可能是DAB 2缺陷细胞中细胞-细胞粘附异常的原因。因此，DAB 2被鉴定为斑珠蛋白结合伴侣，其相互作用由DAB 2的磷酸酪氨酸结合结构域和斑珠蛋白的Asn-Pro-Asp-Tyr（NPDY）基序介导。分子分析和转录组分析还显示，DAB 2参与调节胰岛素样生长因子2介导的信号传导和p53，天冬酰胺合成酶和谷胱甘肽过氧化物酶2的表达。对52个ESC相关miRNAs的表达筛选进一步揭示了DAB 2与细胞死亡、分化和发育相关的信号网络之间的相互作用。因此，这项研究确定了DAB 2在胚胎干细胞命运决定中的作用，并表明存在着一个DAB 2相关的调节回路，控制中胚层分化。J.细胞。225：92-105，2010. (C)2010 Wiley-Liss，Inc.
A variety of signaling networks are implicated in the control of mesoderm differentiation. Previous studies demonstrated that Disabled-2 (DAB2) is a multifunctional protein involved in growth factor signaling and embryonic development. In this study, we investigated DAB2 expression and function during in vitro mesoderm differentiation of murine embryonic stem cells (ESCs). We found that DAB2 was up-regulated when ESCs were co-cultured with OP9 stromal cells for mesoderm differentiation. DAB2 was also up-regulated when ESCs were induced for embryoid body formation. Expression of DAB2 short hairpin small interfering RNA (shDAB2) did not alter the puripotency of ESCs. However, shDAB2 disrupted ESCs cell-cell adhesion and affected embryoid body and colony formation that subsequently impeded mesoderm differentiation of ESCs. Immunofluorescent staining revealed that disorganization of P-catenin and plakoglobin cellular distribution may account for the aberrant cell-cell adhesion in DAB2-deficient cells. Accordingly, DAB2 was identified as a plakoglobin-binding partner with the interaction mediated by the phosphotyrosine binding domain of DAB2 and the Asn-Pro-Asp-Tyr (NPDY) motif of plakoglobin. Molecular analysis and transcriptome profiling also revealed that DAB2 was involved in the regulation of insulin-like growth factor 2-mediated signaling and in the expression of p53, asparagine synthetase and glutathione peroxidase 2. Expression screening of 52 ESCs-related miRNAs further unveiled the interplay between DAB2 and the signaling networks associated with cell death, differentiation and development. This study thereby defines a role of DAB2 in fate determination of ESCs and suggests the presence of a DAB2-associated regulatory circuit in the control of mesoderm differentiation. J. Cell. Physiol. 225: 92-105, 2010. (C) 2010 Wiley-Liss, Inc.