title: Developmental effects of tobacco smoke exposure during human embryonic stem cell differentiation are mediated through the transforming growth factor-β superfamily member, Nodal.
abstract: While the pathologies associated with in utero smoke exposure are well established, their underlying molecular mechanisms are incompletely understood. We differentiated human embryonic stem cells in the presence of physiological concentrations of tobacco smoke and nicotine. Using post hoc microarray analysis, quantitative PCR, and immunoblot analysis, we demonstrated that tobacco smoke has lineage- and stage-specific effects on human embryonic stem cell differentiation, through both nicotine-dependent and -independent pathways. We show that three major stem cell pluripotency/differentiation pathways, Notch, canonical Wnt, and transforming growth factor-β, are affected by smoke exposure, and that Nodal signaling through SMAD2 is specifically impacted by effects on Lefty1, Nodal, and FoxH1. These events are associated with upregulation of microRNA-302a, a post-transcriptional silencer of Lefty1. The described studies provide insight into the mechanisms by which tobacco smoke influences fetal development at the cellular level, and identify specific transcriptional, post-transcriptional, and signaling pathways by which this likely occurs.
authors.affiliation: Cardiovascular Research Institute, University of California, San Francisco, CA 94143-1346, USA.
authors.forename: Walter
authors.initials: W
authors.lastname: Liszewski
authors.forename: Carissa
authors.initials: C
authors.lastname: Ritner
authors.forename: Julian
authors.initials: J
authors.lastname: Aurigui
authors.forename: Sharon S Y
authors.initials: SS
authors.lastname: Wong
authors.forename: Naveed
authors.initials: N
authors.lastname: Hussain
authors.forename: Winfried
authors.initials: W
authors.lastname: Krueger
authors.forename: Cheryl
authors.initials: C
authors.lastname: Oncken
authors.forename: Harold S
authors.initials: HS
authors.lastname: Bernstein
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D055457:Nodal Protein / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D012906:Smoke; D016212:Transforming Growth Factor beta / Q000502:physiology*
chemical_list: C526007:NODAL protein, human; D055457:Nodal Protein; D012906:Smoke; D016212:Transforming Growth Factor beta