Small molecule-based disruption of the Axin/β-catenin protein complex regulates mesenchymal stem cell differentiation
Small molecule-based disruption of the Axin/β-catenin protein complex regulates mesenchymal stem cell differentiation
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DOI：
10.1038/cr.2011.127
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发表时间：
2012-01-01
期刊：
影响因子：
44.1
通讯作者：
Oh, Sangtaek
中科院分区：
文献类型：
--
作者：
Gwak, Jungsug;Hwang, Sun Gwan;Oh, Sangtaek
Wnt/β-连环蛋白通路在包括间充质干细胞在内的多种细胞类型的分化中起重要作用。使用基于细胞的化学筛选分析与270 000化合物的合成化学库，我们确定了化合物SKL 2001作为Wnt/β-连环蛋白途径的新型激动剂，并揭示了其分子作用机制。SKL 2001通过增加细胞内β-catenin蛋白水平上调β-catenin应答性转录，并抑制β-catenin残基Ser 33/37/Thr 41和Ser 45的磷酸化，这将标志着它的蛋白酶体降解，而不影响CK 1和GSK-3 β酶活性。生化分析显示，SKL 2001破坏了Axin/β-catenin相互作用，这是CK 1和GSK-3 β介导的β-catenin在Ser 33/37/Thr 41和Ser 45磷酸化的关键步骤。用SKL 2001处理间充质干细胞促进成骨细胞生成并抑制脂肪细胞分化，这两者都伴随着Wnt/β-catenin通路的激活。我们的研究结果提供了一种新的策略，通过调节Wnt/β-连环蛋白通路来调节间充质干细胞分化。
The Wnt/beta-catenin pathway plays important roles in the differentiation of multiple cell types, including mesenchymal stem cells. Using a cell-based chemical screening assay with a synthetic chemical library of 270 000 compounds, we identified the compound SKL2001 as a novel agonist of the Wnt/beta-catenin pathway and uncovered its molecular mechanism of action. SKL2001 upregulated beta-catenin responsive transcription by increasing the intracellular beta-catenin protein level and inhibited the phosphorylation of beta-catenin at residues Ser33/37/Thr41 and Ser45, which would mark it for proteasomal degradation, without affecting CK1 and GSK-3 beta enzyme activities. Biochemical analysis revealed that SKL2001 disrupted the Axin/beta-catenin interaction, which is a critical step for CK1- and GSK-3 beta-mediated phosphorylation of beta-catenin at Ser33/37/Thr41 and Ser45. The treatment of mesenchymal stem cells with SKL2001 promoted osteoblastogenesis and suppressed adipocyte differentiation, both of which were accompanied by the activation of Wnt/beta-catenin pathway. Our findings provide a new strategy to regulate mesenchymal stem cell differentiation by modulation of the Wnt/beta-catenin pathway.