Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling
Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling
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DOI：
10.1016/j.yjmcc.2014.12.003
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发表时间：
2015-03-01
影响因子：
5
通讯作者：
Oh, Steve K. W.
中科院分区：
文献类型：
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作者：
Laco, Filip;Low, Joo-Leng;Oh, Steve K. W.
人类多能干细胞分化为类胚体(EBS)以前曾用p38alfa MAPK抑制剂如SB203580实现，中等效率为10-15%。我们合成并筛选了42个化合物，它们是SB203580的2，4，5-三取代唑类似物，用于有效地诱导心肌细胞分化。我们的筛选发现了与SB203580具有相似心脏分化活性的新化合物。然而，心脏分化与p38alfa MAPK抑制无关，这表明心脏分化存在另一种机制。在分析了几个2，4，5-三取代唑化合物对97个蛋白激酶的作用后，我们确定了几个关闭的靶标，其中包括酪蛋白激酶1(CK1)。SB203580及其类似物的生心肌活性与中胚层后Wnt/β-catenin通路对CK1、epsilon和Delta的抑制有关。这些发现将2，4，5-三取代唑的作用机制与Wnt/β-catenin调控心脏分化途径的当前理论相结合。因此，用Wnt激活剂CHIR99021和2，4，5-三取代唑开发了一种有效的心肌细胞方案，使心肌细胞的产量达到50-70%，生长速度增加了2倍。(C)2014爱思唯尔有限公司。保留所有权利。
Differentiation of human pluripotent stem cells as embryoid bodies (EBs) has been achieved previously with p38alfa MAPK inhibitors such as SB203580 with moderate efficiency of 10-15%. We synthesized and screened 42 compounds that are 2,4,5-trisubstituted azole analogues of SB203580 for efficient cardiomyocyte differentiation. Our screen identified novel compounds that have similar cardiac differentiation activity as SB203580. However, the cardiac differentiation did not correlate with p38alfa MAPK inhibition, indicating an alternative mechanism in cardiac differentiation. Upon profiling several 2,4,5-trisubstituted azole compounds against a panel of 97 kinases we identified several off targets, among them casein kinases 1 (CK1). The cardiomyogenic activities of SB203580 and its analogues showed a correlation with post mesoderm Wnt/beta-catenin pathway inhibition of CK1 epsilon and delta. These findings united the mechanism of 2,4,5-trisubstituted azole with the current theory of Wnt/beta-catenin regulated pathway of cardiac differentiation. Consequently an efficient cardiomyocyte protocol was developed with Wnt activator CHIR99021 and 2,4,5-trisubstituted azoles to give high yields of 50-70% cardiomyocytes and a 2-fold increase in growth. (C) 2014 Elsevier Ltd. All rights reserved.