title: Wnt Signaling Regulates Airway Epithelial Stem Cells in Adult Murine Submucosal Glands.
abstract: Wnt signaling is required for lineage commitment of glandular stem cells (SCs) during tracheal submucosal gland (SMG) morphogenesis from the surface airway epithelium (SAE). Whether similar Wnt-dependent processes coordinate SC expansion in adult SMGs following airway injury remains unknown. We found that two Wnt-reporters in mice (BAT-gal and TCF/Lef:H2B-GFP) are coexpressed in actively cycling SCs of primordial glandular placodes and in a small subset of adult SMG progenitor cells that enter the cell cycle 24 hours following airway injury. At homeostasis, these Wnt reporters showed nonoverlapping cellular patterns of expression in the SAE and SMGs. Following tracheal injury, proliferation was accompanied by dynamic changes in Wnt-reporter activity and the analysis of 56 Wnt-related signaling genes revealed unique temporal changes in expression within proximal (gland-containing) and distal (gland-free) portions of the trachea. Wnt stimulation in vivo and in vitro promoted epithelial proliferation in both SMGs and the SAE. Interestingly, slowly cycling nucleotide label-retaining cells (LRCs) of SMGs were spatially positioned near clusters of BAT-gal positive serous tubules. Isolation and culture of tet-inducible H2B-GFP LRCs demonstrated that SMG LRCs were more proliferative than SAE LRCs and culture expanded SMG-derived progenitor cells outcompeted SAE-derived progenitors in regeneration of tracheal xenograft epithelium using a clonal analysis competition assay. SMG-derived progenitors were also multipotent for cell types in the SAE and formed gland-like structures in xenografts. These studies demonstrate the importance of Wnt signals in modulating SC phenotypes within tracheal niches and provide new insight into phenotypic differences of SMG and SAE SCs. Stem Cells 2016;34:2758-2771.
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Lynch
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Preston J
authors.initials: PJ
authors.lastname: Anderson
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.|Molecular and Cellular Biology Program, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Weiliang
authors.initials: W
authors.lastname: Xie
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Adrianne K
authors.initials: AK
authors.lastname: Crooke
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.|Center for Gene Therapy, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.|Molecular and Cellular Biology Program, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Scott R
authors.initials: SR
authors.lastname: Tyler
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Meihui
authors.initials: M
authors.lastname: Luo
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Kusner
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, USA.
authors.forename: Traci
authors.initials: T
authors.lastname: Neff
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Burnette
authors.affiliation: Central Microscopy Research Facility, Iowa City, Iowa, USA.
authors.forename: Katherine S
authors.initials: KS
authors.lastname: Walters
authors.affiliation: Department of Obstetrics and Gynecology, University of Iowa Hospitals and Clinics, Iowa City, Iowa, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Goodheart
authors.affiliation: Department of Cardiothoracic Surgery, University of Iowa Carver College of Medicine, Iowa City, Iowa, USA.
authors.forename: Kalpaj R
authors.initials: KR
authors.lastname: Parekh
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.|Center for Gene Therapy, University of Iowa, Iowa City, Iowa, USA.
authors.forename: John F
authors.initials: JF
authors.lastname: Engelhardt
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005088:Exocrine Glands / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D064593:Heterografts; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D019394:Ki-67 Antigen / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009281:Naphthalenes / Q000633:toxicity; D009940:Organoids / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D061251:Primary Cell Culture; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D046509:Tissue Culture Techniques; D014132:Trachea / Q000187:drug effects / Q000293:injuries / Q000378:metabolism* / Q000601:surgery; D051155:Wnt1 Protein / Q000235:genetics / Q000378:metabolism*; D060509:Wnt3A Protein / Q000235:genetics / Q000378:metabolism*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: C501127:Dkk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D019394:Ki-67 Antigen; C498355:Mki67 protein, mouse; D009281:Naphthalenes; D051155:Wnt1 Protein; C495246:Wnt1 protein, mouse; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; D049452:Green Fluorescent Proteins; C031721:naphthalene; D001616:beta-Galactosidase
keywords: Wnt signaling pathway; adult stem cells; developmental biology; multipotential differentiation; stem cell niche; stem cell-microenvironment interactions; trachea