title: Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling.
abstract: 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.
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore.
authors.forename: Filip
authors.initials: F
authors.lastname: Laco
authors.affiliation: Institute of Chemical and Engineering Sciences, 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore.
authors.forename: Joo-Leng
authors.initials: JL
authors.lastname: Low
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Seow
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore.
authors.forename: Tsung Liang
authors.initials: TL
authors.lastname: Woo
authors.affiliation: Department of Pharmacy, Faculty of Science, National University of Singapore, 18 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Qixing
authors.initials: Q
authors.lastname: Zhong
authors.affiliation: Institute of Chemical and Engineering Sciences, 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore.
authors.forename: Jayasree
authors.initials: J
authors.lastname: Seayad
authors.affiliation: National Heart Research Institute Singapore, National Heart Centre Singapore, Singapore; Cardiovascular & Metabolic Disorders Program, DUKE-NUS Graduate Medical School Singapore, Singapore.
authors.forename: Zhenfeng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: National Heart Research Institute Singapore, National Heart Centre Singapore, Singapore.
authors.forename: Heiming
authors.initials: H
authors.lastname: Wei
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore.
authors.forename: Shaul
authors.initials: S
authors.lastname: Reuveny
authors.affiliation: Murdoch Childrens Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, Melbourne, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Elliott
authors.affiliation: Institute of Chemical and Engineering Sciences, 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore; Department of Pharmacy, Faculty of Science, National University of Singapore, 18 Science Drive 4, Singapore 117543, Singapore.
authors.forename: Christina L L
authors.initials: CL
authors.lastname: Chai
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore. Electronic address: steve_oh@bti.a-star.edu.sg.
authors.forename: Steve K W
authors.initials: SK
authors.lastname: Oh
mesh_terms: D000818:Animals; D047389:Casein Kinase I / Q000037:antagonists & inhibitors*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D015195:Drug Design; D006801:Humans; D007093:Imidazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D038081:Organogenesis / Q000187:drug effects; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology; D011725:Pyridines / Q000138:chemical synthesis / Q000494:pharmacology*; D060449:Wnt Signaling Pathway / Q000187:drug effects*
chemical_list: D007093:Imidazoles; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D047389:Casein Kinase I; C093642:SB 203580
keywords: Cardiomyocyte differentiation; Casein kinase 1; Kinase inhibitors; Stem cell; Wnt signaling pathway; p38 MAPK