title: In situ mechanotransduction via vinculin regulates stem cell differentiation.
abstract: Human mesenchymal stem cell (hMSC) proliferation, migration, and differentiation have all been linked to extracellular matrix stiffness, yet the signaling pathway(s) that are necessary for mechanotransduction remain unproven. Vinculin has been implicated as a mechanosensor in vitro, but here we demonstrate its ability to also regulate stem cell behavior, including hMSC differentiation. RNA interference-mediated vinculin knockdown significantly decreased stiffness-induced MyoD, a muscle transcription factor, but not Runx2, an osteoblast transcription factor, and impaired stiffness-mediated migration. A kinase binding accessibility screen predicted a cryptic MAPK1 signaling site in vinculin which could regulate these behaviors. Indeed, reintroduction of vinculin domains into knocked down cells indicated that MAPK1 binding site-containing vinculin constructs were necessary for hMSC expression of MyoD. Vinculin knockdown does not appear to interfere with focal adhesion assembly, significantly alter adhesive properties, or diminish cell traction force generation, indicating that its knockdown only adversely affected MAPK1 signaling. These data provide some of the first evidence that a force-sensitive adhesion protein can regulate stem cell fate.
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, California, USA.
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Holle
authors.forename: Xinyi
authors.initials: X
authors.lastname: Tang
authors.forename: Deepthi
authors.initials: D
authors.lastname: Vijayraghavan
authors.forename: Ludovic G
authors.initials: LG
authors.lastname: Vincent
authors.forename: Alexander
authors.initials: A
authors.lastname: Fuhrmann
authors.forename: Yu Suk
authors.initials: YS
authors.lastname: Choi
authors.forename: Juan C
authors.initials: JC
authors.lastname: del Álamo
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Engler
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D020935:MAP Kinase Signaling System; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D018625:Microscopy, Atomic Force / Q000379:methods; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D010766:Phosphorylation; D016608:Talin / Q000378:metabolism; D014162:Transfection; D016596:Vinculin / Q000378:metabolism*
chemical_list: D016608:Talin; D016596:Vinculin; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1
keywords: Cellular mechanotransduction; Differentiation; Mesenchymal stem cell; Myogenesis
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