title: Efficient Derivation of Functional Human Airway Epithelium from Pluripotent Stem Cells via Temporal Regulation of Wnt Signaling.
abstract: Effective derivation of functional airway organoids from induced pluripotent stem cells (iPSCs) would provide valuable models of lung disease and facilitate precision therapies for airway disorders such as cystic fibrosis. However, limited understanding of human airway patterning has made this goal challenging. Here, we show that cyclical modulation of the canonical Wnt signaling pathway enables rapid directed differentiation of human iPSCs via an NKX2-1+progenitor intermediate into functional proximal airway organoids. We find that human NKX2-1+progenitors have high levels of Wnt activation but respond intrinsically to decreases in Wnt signaling by rapidly patterning into proximal airway lineages at the expense of distal fates. Using this directed approach, we were able to generate cystic fibrosis patient-specific iPSC-derived airway organoids with a defect in forskolin-induced swelling that is rescued by gene editing to correct the disease mutation. Our approach has many potential applications in modeling and drug screening for airway diseases.
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Katherine B
authors.initials: KB
authors.lastname: McCauley
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Finn
authors.initials: F
authors.lastname: Hawkins
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Serra
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA.
authors.forename: Dylan C
authors.initials: DC
authors.lastname: Thomas
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Anjali
authors.initials: A
authors.lastname: Jacob
authors.affiliation: Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA. Electronic address: dkotton@bu.edu.
authors.forename: Darrell N
authors.initials: DN
authors.lastname: Kotton
mesh_terms: D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009940:Organoids / Q000166:cytology / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism*; D000074482:Thyroid Nuclear Factor 1 / Q000378:metabolism*; D060449:Wnt Signaling Pathway
chemical_list: C000614412:NKX2-1 protein, human; D000074482:Thyroid Nuclear Factor 1
keywords: Wnt signaling; airway epithelium; cystic fibrosis; directed differentiation; disease modeling; human pluripotent stem cells; lung development; lung epithelium