Wnt/β-Catenin Signaling Blockade Promotes Neuronal Induction and Dopaminergic Differentiation in Embryonic Stem Cells
Wnt/β-Catenin Signaling Blockade Promotes Neuronal Induction and Dopaminergic Differentiation in Embryonic Stem Cells
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DOI：
10.1002/stem.210
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发表时间：
2009-12-01
期刊：
影响因子：
5.2
通讯作者：
Arenas, Ernest
中科院分区：
文献类型：
--
作者：
Cajanek, Lukas;Ribeiro, Diogo;Arenas, Ernest
胚胎干细胞（ESCs）不仅是细胞替代疗法的一个有前途的细胞来源，而且是研究细胞信号传导和多巴胺能（DA）神经元发育的分子机制的工具。DA神经元发育的主要调节因子之一是Wnt信号。在这里，我们使用缺乏Wnt 1或低密度脂蛋白受体相关蛋白6（LRP 6）的小鼠胚胎干细胞（mESC）来破译Wnt/β-连环蛋白信号传导对mESC中DA神经元发育的作用。我们提供的证据表明，LRP 6的缺失废除了mESC对Wnt配体刺激的反应性。使用两种分化方案，我们表明，Wnt 1或LRP 6的损失增加神经外胚层分化和mESC衍生的DA神经元的数量。这些作用与用Wnt/β-连环蛋白通路抑制剂Dickkopf 1（Dkk 1）处理mESC后观察到的作用相似。结合，我们的结果表明，Wnt/β-连环蛋白信号的减少增强mESC的神经元和DA分化。这些研究结果表明：1）Wnt 1或LRP 6不是mESC体外DA分化的严格要求，2）需要优化ESC培养物中形态发生素的水平及其活性以改善DA分化，以及3）通过用Dkk 1增强ESC衍生的DA神经元的分化和数量，可以改善ESC用于帕金森病的细胞替代疗法的应用。干细胞2009;27：2917-2927
Embryonic stem cells (ESCs) represent not only a promising source of cells for cell replacement therapy, but also a tool to study the molecular mechanisms underlying cellular signaling and dopaminergic (DA) neuron development. One of the main regulators of DA neuron development is Wnt signaling. Here we used mouse ESCs (mESCs) lacking Wnt1 or the low-density lipoprotein receptor-related protein 6 (LRP6) to decipher the action of Wnt/beta-catenin signaling on DA neuron development in mESCs. We provide evidence that the absence of LRP6 abrogates responsiveness of mESCs to Wnt ligand stimulation. Using two differentiation protocols, we show that the loss of Wnt1 or LRP6 increases neuroectodermal differentiation and the number of mESC-derived DA neurons. These effects were similar to those observed following treatment of mESCs with the Wnt/beta-catenin pathway inhibitor Dickkopf1 (Dkk1). Combined, our results show that decreases in Wnt/beta-catenin signaling enhance neuronal and DA differentiation of mESCs. These findings suggest that: 1) Wnt1 or LRP6 are not strictly required for the DA differentiation of mESCs in vitro, 2) the levels of morphogens and their activity in ESC cultures need to be optimized to improve DA differentiation, and 3) by enhancing the differentiation and number of ESC-derived DA neurons with Dkk1, the application of ESCs for cell replacement therapy in Parkinson's disease may be improved. STEM CELLS 2009;27:2917-2927