title: Identification of Small Molecules Which Induce Skeletal Muscle Differentiation in Embryonic Stem Cells via Activation of the Wnt and Inhibition of Smad2/3 and Sonic Hedgehog Pathways.
abstract: The multilineage differentiation capacity of mouse and human embryonic stem (ES) cells offers a testing platform for small molecules that mediate mammalian lineage determination and cellular specialization. Here we report the identification of two small molecules which drives mouse 129 ES cell differentiation to skeletal muscle with high efficiency without any genetic modification. Mouse embryoid bodies (EBs) were used to screen a library of 1,000 small molecules to identify compounds capable of inducing high levels of Pax3 mRNA. Stimulation of EBs with SMIs (skeletal muscle inducer, SMI1 and SMI2) from the screen resulted in a high percentage of intensively twitching skeletal muscle fibers 3 weeks after induction. Gene expression profiling studies that were carried out for mode of actions analysis showed that SMIs activated genes regulated by the Wnt pathway and inhibited expression of Smad2/3 and Sonic Hedgehog (Shh) target genes. A combination of three small molecules known to modulate these three pathways acted similarly to the SMIs found here, driving ES cells from 129 as well as Balb/c and C57Bl/6 to skeletal muscle. Taken together, these data demonstrate that the SMI drives ES cells to skeletal muscle via concerted activation of the Wnt pathway, and inhibition of Smad2/3 signaling and Shh pathways. This provides important developmental biological information about skeletal muscle differentiation from embryonic stem cells and may lead to the development of new therapeutics for muscle disease.
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.|Faculty of Sciences, University of Basel, Basel, Switzerland.
authors.forename: Hyunwoo
authors.initials: H
authors.lastname: Lee
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Corinne
authors.initials: C
authors.lastname: Haller
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Carole
authors.initials: C
authors.lastname: Manneville
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Thierry
authors.initials: T
authors.lastname: Doll
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Fruh
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Caroline Gubser
authors.initials: CG
authors.lastname: Keller
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Shola M
authors.initials: SM
authors.lastname: Richards
authors.affiliation: Center for Proteomic Chemistry, Basel, Switzerland.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Ibig-Rehm
authors.affiliation: Global Discovery Chemistry, Novartis Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Maude
authors.initials: M
authors.lastname: Patoor
authors.affiliation: Global Discovery Chemistry, Novartis Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Marjo
authors.initials: M
authors.lastname: Goette
authors.affiliation: Global Discovery Chemistry, Novartis Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Laure C
authors.initials: LC
authors.lastname: Bouchez
authors.affiliation: Developmental and Molecular Pathways, Basel, Switzerland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mueller
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D053823:Hedgehog Proteins / Q000378:metabolism*; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018656:Muscle Fibers, Fast-Twitch / Q000166:cytology / Q000378:metabolism*; D051899:Smad2 Protein / Q000378:metabolism*; D051900:Smad3 Protein / Q000378:metabolism*; D060449:Wnt Signaling Pathway
chemical_list: D053823:Hedgehog Proteins; C465570:Shh protein, mouse; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D051900:Smad3 Protein; C497791:Smad3 protein, mouse
keywords: Cellular therapy; Differentiation; Embryonic stem cells; Mesoderm; Muscle stem cells; Myogenesis; Skeletal muscle