title: Noncanonical NOTCH signaling limits self-renewal of human epithelial and induced pluripotent stem cells through ROCK activation.
abstract: NOTCH plays essential roles in cell fate specification during embryonic development and in adult tissue maintenance. In keratinocytes, it is a key inducer of differentiation. ROCK, an effector of the small GTPase Rho, is also implicated in keratinocyte differentiation, and its inhibition efficiently potentiates immortalization of human keratinocytes and greatly improves survival of dissociated human pluripotent stem cells. However, the molecular basis for ROCK activation is not fully established in these contexts. Here we provide evidence that intracellular forms of NOTCH1 trigger the immediate activation of ROCK1 independent of its transcriptional activity, promoting differentiation and resulting in decreased clonogenicity of normal human keratinocytes. Knockdown of NOTCH1 abrogated ROCK1 activation and conferred sustained clonogenicity upon differentiation stimuli. Treatment with a ROCK inhibitor, Y-27632, or ROCK1 silencing substantially rescued the growth defect induced by activated NOTCH1. Furthermore, we revealed that impaired self-renewal of human induced pluripotent stem cells upon dissociation is, at least in part, attributable to NOTCH-dependent ROCK activation. Thus, the present study unveils a novel NOTCH-ROCK pathway critical for cellular differentiation and loss of self-renewal capacity in a subset of immature cells.
authors.affiliation: Division of Virology, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yugawa
authors.forename: Koichiro
authors.initials: K
authors.lastname: Nishino
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Ohno
authors.forename: Tomomi
authors.initials: T
authors.lastname: Nakahara
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Fujita
authors.forename: Naoki
authors.initials: N
authors.lastname: Goshima
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
authors.forename: Tohru
authors.initials: T
authors.lastname: Kiyono
mesh_terms: D000328:Adult; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004789:Enzyme Activation; D005260:Female; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: C497761:NOTCH1 protein, human; D051881:Receptor, Notch1; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases
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