title: Small molecule-based disruption of the Axin/β-catenin protein complex regulates mesenchymal stem cell differentiation.
abstract: The Wnt/β-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/β-catenin pathway and uncovered its molecular mechanism of action. SKL2001 upregulated β-catenin responsive transcription by increasing the intracellular β-catenin protein level and inhibited the phosphorylation of β-catenin at residues Ser33/37/Thr41 and Ser45, which would mark it for proteasomal degradation, without affecting CK1 and GSK-3β enzyme activities. Biochemical analysis revealed that SKL2001 disrupted the Axin/β-catenin interaction, which is a critical step for CK1- and GSK-3β-mediated phosphorylation of β-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/β-catenin pathway. Our findings provide a new strategy to regulate mesenchymal stem cell differentiation by modulation of the Wnt/β-catenin pathway.
authors.affiliation: Department of Advanced Fermentation Fusion Science & Technology, Kookmin University, Seoul, Korea.
authors.forename: Jungsug
authors.initials: J
authors.lastname: Gwak
authors.forename: Sun Gwan
authors.initials: SG
authors.lastname: Hwang
authors.forename: Hyung-Soon
authors.initials: HS
authors.lastname: Park
authors.forename: Sang Rak
authors.initials: SR
authors.lastname: Choi
authors.forename: Sun-Hee
authors.initials: SH
authors.lastname: Park
authors.forename: Hyunjoon
authors.initials: H
authors.lastname: Kim
authors.forename: Nam-Chul
authors.initials: NC
authors.lastname: Ha
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Bae
authors.forename: Jin-Kwan
authors.initials: JK
authors.lastname: Han
authors.forename: Dong-Eun
authors.initials: DE
authors.lastname: Kim
authors.forename: Jeong Woo
authors.initials: JW
authors.lastname: Cho
authors.forename: Sangtaek
authors.initials: S
authors.lastname: Oh
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D060465:Axin Signaling Complex / Q000378:metabolism*; D002454:Cell Differentiation; D004789:Enzyme Activation; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D057809:HEK293 Cells; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D047468:Immunoprecipitation; D007555:Isoxazoles / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D025941:Protein Interaction Mapping; D059748:Proteolysis; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D014162:Transfection; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000819:agonists / Q000378:metabolism*
chemical_list: C571720:5-furan-2yl-isoxazole-3-carboxylic acid (3-imidazol-1yl-propyl)-amide; D060465:Axin Signaling Complex; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007555:Isoxazoles; D051176:beta Catenin; D013912:Threonine; D012694:Serine; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3