Endogenous Wnt/beta-catenin signaling is required for cardiac differentiation in human embryonic stem cells.
Endogenous Wnt/beta-catenin signaling is required for cardiac differentiation in human embryonic stem cells.
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DOI：
10.1371/journal.pone.0011134
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发表时间：
2010-06-15
期刊：
影响因子：
3.7
通讯作者：
Murry CE
中科院分区：
文献类型：
--
作者：
Paige SL;Osugi T;Afanasiev OK;Pabon L;Reinecke H;Murry CE
Wnt/β-catenin信号传导是分化和形态发生的重要调节因子，也可以控制干细胞命运。我们的小组已经开发了一种有效的方案，通过激活素A和BMP 4诱导从人胚胎干细胞（ES细胞）产生心肌细胞。我们测试了Wnt/β-catenin信号控制该系统中早期中胚层诱导和后期心脏分化的假设。在诱导时添加外源性Wnt 3a增强了心脏分化，而用Dkk 1早期抑制内源性Wnt/β-catenin信号传导抑制了心脏分化，如β-肌球蛋白重链（β-MHC）、心肌肌钙蛋白T（cTnT）、Nkx 2.5的定量RT-PCR分析和肌节肌球蛋白重链（sMHC）的流式细胞术分析所示。相反，内源性产生的Wnt的晚期拮抗作用增强了心脏发生，表明该途径在人类心脏分化中的双相作用。使用定量RT-PCR，我们表明，典型的Wnt配体的表达诱导激活素A/BMP 4处理，早期Wnt配体表达的程度可以预测随后的心脏发生的效率。Brachyury表达的测量显示，添加Wnt 3a增强中胚层诱导，而阻断内源性产生的Wnt显著抑制中胚层形成。最后，我们发现Wnt/β-catenin信号是BMP 4激活Smad 1所必需的。我们的数据表明，从人ES细胞诱导中胚层和随后的心脏分化需要激活素A/BMP 4和Wnt/β-连环蛋白通路之间的微调串扰。控制这些途径可以有效地产生用于基础研究或心脏修复应用的心肌细胞。
Wnt/β-catenin signaling is an important regulator of differentiation and morphogenesis that can also control stem cell fates. Our group has developed an efficient protocol to generate cardiomyocytes from human embryonic stem (ES) cells via induction with activin A and BMP4. We tested the hypothesis that Wnt/β-catenin signals control both early mesoderm induction and later cardiac differentiation in this system. Addition of exogenous Wnt3a at the time of induction enhanced cardiac differentiation, while early inhibition of endogenous Wnt/β-catenin signaling with Dkk1 inhibited cardiac differentiation, as indicated by quantitative RT-PCR analysis for β-myosin heavy chain (β-MHC), cardiac troponin T (cTnT), Nkx2.5, and flow cytometry analysis for sarcomeric myosin heavy chain (sMHC). Conversely, late antagonism of endogenously produced Wnts enhanced cardiogenesis, indicating a biphasic role for the pathway in human cardiac differentiation. Using quantitative RT-PCR, we show that canonical Wnt ligand expression is induced by activin A/BMP4 treatment, and the extent of early Wnt ligand expression can predict the subsequent efficiency of cardiogenesis. Measurement of Brachyury expression showed that addition of Wnt3a enhances mesoderm induction, whereas blockade of endogenously produced Wnts markedly inhibits mesoderm formation. Finally, we show that Wnt/β-catenin signaling is required for Smad1 activation by BMP4. Our data indicate that induction of mesoderm and subsequent cardiac differentiation from human ES cells requires fine-tuned cross talk between activin A/BMP4 and Wnt/β-catenin pathways. Controlling these pathways permits efficient generation of cardiomyocytes for basic studies or cardiac repair applications.
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DOI：
10.1073/pnas.0704044104
发表时间：
2007-06-26
影响因子：
11.1
作者：
Kwon, Chulan;Arnold, Joshua;Srivastava, Deepak
通讯作者：
Srivastava, Deepak
DOI：
10.1073/pnas.0605768103
发表时间：
2006-12-26
影响因子：
11.1
作者：
Naito, Atsuhiko T.;Shiojima, Ichiro;Komuro, Issei
通讯作者：
Komuro, Issei
影响因子：
2.5
作者：
Golob, Jonathan L.;Paige, Sharon L.;Murry, Charles E.
通讯作者：
Murry, Charles E.
影响因子：
15.9
作者：
Kehat, I;Kenyagin-Karsenti, D;Gepstein, L
通讯作者：
Gepstein, L
影响因子：
10.5
作者：
Schneider, VA;Mercola, M
通讯作者：
Mercola, M