GRHL2 coordinates regeneration of a polarized mucociliary epithelium from basal stem cells
Reads0
Chats0
TLDR
Crispr/Cas9-mediated mutation of the transcription factor GRHL2 or either of its predicted downstream targets ZNF750 and SMAGP in primary human bronchial epithelial basal cells leads to defects in ciliogenesis and/or barrier function.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.read more
Citations
More filters
Journal ArticleDOI
Lung organoids: current uses and future promise.
Christina E. Barkauskas,Mei-I Chung,Bryan A. Fioret,Xia Gao,Hiroaki Katsura,Brigid L.M. Hogan +5 more
TL;DR: This Review summarizes the different methods for generating organoids from cells isolated from human and mouse lungs, and compares their final structure and cellular composition with that of the airways or alveoli of the adult lung.
Journal ArticleDOI
Organoid technology and applications in cancer research
TL;DR: In conclusion, organoids represent an excellent preclinical model for human tumors, promoting the translation from basic cancer research to clinical practice and its applications in cancer research are summarized.
Journal ArticleDOI
Building and Regenerating the Lung Cell by Cell
TL;DR: The unique architecture of the mammalian lung is required for adaptation to air breathing at birth and thereafter as discussed by the authors, and the cellular and molecular mechanisms controlling its morphogenesis are discussed in detail.
Journal ArticleDOI
Dissecting the cellular specificity of smoking effects and reconstructing lineages in the human airway epithelium.
K.C. Goldfarbmuren,Nathan D. Jackson,Satria Sajuthi,Nathan Dyjack,Katie S. Li,Cydney Rios,Elizabeth G. Plender,M.T. Montgomery,Jamie L. Everman,Preston E. Bratcher,Eszter K. Vladar,Max A. Seibold +11 more
TL;DR: Single-cell RNA sequencing is used to map cell type-specific changes in human tracheal epithelium related to smoking, and to provide evidence for a tuft-like progenitor for pulmonary neuroendocrine cells and ionocytes.
Journal ArticleDOI
Human lung development: recent progress and new challenges.
TL;DR: Current understanding of human lung development has been profoundly influenced by studies in mice and, more recently, by experiments using in vitro human lung developmental models and RNA sequencing of human foetal lung tissue.
References
More filters
Journal ArticleDOI
Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells
Ophir Shalem,Ophir Shalem,Neville E. Sanjana,Neville E. Sanjana,Ella Hartenian,Xi-Shun Shi,David A. Scott,David A. Scott,Tarjei S. Mikkelsen,Dirk Heckl,Benjamin L. Ebert,David E. Root,John G. Doench,Feng Zhang,Feng Zhang +14 more
TL;DR: This work shows that lentiviral delivery of a genome-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes with 64,751 unique guide sequences enables both negative and positive selection screening in human cells, and observes a high level of consistency between independent guide RNAs targeting the same gene and a high rate of hit confirmation.
Journal ArticleDOI
Improved vectors and genome-wide libraries for CRISPR screening.
TL;DR: In this paper, Zhang et al. used a Genome-scale CRISPR Knock-Out (GeCKO) library to identify loss-of-function mutations in a melanoma model.
Journal ArticleDOI
Organogenesis in a dish: modeling development and disease using organoid technologies.
TL;DR: These studies illustrated two key events in structural organization during organogenesis: cell sorting out and spatially restricted lineage commitment, which are recapitulated in organoids, which self-assemble to form the cellular organization of the organ itself.
Journal ArticleDOI
Basal cells as stem cells of the mouse trachea and human airway epithelium
Jason R. Rock,Mark W. Onaitis,Emma L. Rawlins,Yun Lu,Cheryl P. Clark,Yan Xue,Scott H. Randell,Brigid L.M. Hogan +7 more
TL;DR: The pseudostratified epithelium of the mouse trachea and human airways contains a population of basal cells expressing Trp-63 and cytokeratins 5, which generate differentiated cells during postnatal growth and in the adult during both steady state and epithelial repair.
Journal ArticleDOI
Multiple stromal populations contribute to pulmonary fibrosis without evidence for epithelial to mesenchymal transition
Jason R. Rock,Christina E. Barkauskas,Michael J. Cronce,Yan Xue,Jeffrey R. Harris,Jiurong Liang,Paul W. Noble,Brigid L.M. Hogan +7 more
TL;DR: A previously unappreciated heterogeneity of cell types proliferating in fibrotic lesions is shown and pericytes and two epithelial cell populations are excluded as the origin of myofibroblasts.
Related Papers (5)
Repair and regeneration of the respiratory system: complexity, plasticity, and mechanisms of lung stem cell function.
Brigid L.M. Hogan,Christina E. Barkauskas,Harold A. Chapman,Jonathan A. Epstein,Rajan Jain,Connie C. W. Hsia,Laura E. Niklason,Elizabeth A. Calle,Andrew V. Le,Scott H. Randell,Jason R. Rock,Melinda Snitow,Matthew F. Krummel,Barry R. Stripp,Thiennu H. Vu,Eric S. White,Jeffrey A. Whitsett,Edward E. Morrisey +17 more