title: GRHL2 coordinates regeneration of a polarized mucociliary epithelium from basal stem cells.
abstract: Pseudostratified airway epithelium of the lung is composed of polarized ciliated and secretory cells maintained by basal stem/progenitor cells. An important question is how lineage choice and differentiation are coordinated with apical-basal polarity and epithelial morphogenesis. Our previous studies indicated a key integrative role for the transcription factor Grainyhead-like 2 (Grhl2). In this study, we present further evidence for this model using conditional gene deletion during the regeneration of airway epithelium and clonal organoid culture. We also use CRISPR/Cas9 genome editing in primary human basal cells differentiating into organoids and mucociliary epithelium in vitro. Loss of Grhl2 inhibits organoid morphogenesis and the differentiation of ciliated cells and reduces the expression of both notch and ciliogenesis genes (Mcidas, Rfx2, and Myb) with distinct Grhl2 regulatory sites. The genome editing of other putative target genes reveals roles for zinc finger transcription factor Znf750 and small membrane adhesion glycoprotein in promoting ciliogenesis and barrier function as part of a network of genes coordinately regulated by Grhl2.
authors.affiliation: Department of Cell Biology, Duke University School of Medicine, Durham, NC 27710.
authors.forename: Xia
authors.initials: X
authors.lastname: Gao
authors.affiliation: Department of Cell Biology, Duke University School of Medicine, Durham, NC 27710.
authors.forename: Aman S
authors.initials: AS
authors.lastname: Bali
authors.affiliation: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Scott H
authors.initials: SH
authors.lastname: Randell
authors.affiliation: Department of Cell Biology, Duke University School of Medicine, Durham, NC 27710 brigid.hogan@duke.edu.
authors.forename: Brigid L M
authors.initials: BL
authors.lastname: Hogan
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000502:physiology; D004268:DNA-Binding Proteins / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism* / Q000502:physiology*; D006801:Humans; D008168:Lung / Q000378:metabolism / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009024:Morphogenesis / Q000502:physiology; D012038:Regeneration / Q000502:physiology*; D013234:Stem Cells / Q000378:metabolism* / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism*; D016335:Zinc Fingers / Q000502:physiology
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors