Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells

Hongmei Mou; Vladimir Vinarsky; Purushothama Rao Tata; Karissa Brazauskas; Soon H Choi; Adrianne K Crooke; Bing Zhang; George M Solomon; Brett Turner; Hermann Bihler; Jan Harrington; Allen Lapey; Colleen Channick; Colleen Keyes; Adam Freund; Steven Artandi; Martin Mense; Steven Rowe; John F Engelhardt; Ya-Chieh Hsu; Jayaraj Rajagopal

doi:10.1016/j.stem.2016.05.012

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Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells

Journal article

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Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells

Hongmei Mou, Vladimir Vinarsky, Purushothama Rao Tata, Karissa Brazauskas, Soon H Choi, Adrianne K Crooke, Bing Zhang, George M Solomon, Brett Turner, Hermann Bihler, …

Cell stem cell, Vol.19(2), pp.217-231

08/04/2016

DOI: 10.1016/j.stem.2016.05.012

PMCID: PMC4975684

PMID: 27320041

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Abstract

Functional modeling of many adult epithelia is limited by the difficulty in maintaining relevant stem cell populations in culture. Here, we show that dual inhibition of SMAD signaling pathways enables robust expansion of primary epithelial basal cell populations. We find that TGFβ/BMP/SMAD pathway signaling is strongly activated in luminal and suprabasal cells of several epithelia, but suppressed in p63+ basal cells. In airway epithelium, SMAD signaling promotes differentiation, and its inhibition leads to stem cell hyperplasia. Using dual SMAD signaling inhibition in a feeder-free culture system, we have been able to expand airway basal stem cells from multiple species. Expanded cells can produce functional airway epithelium physiologically responsive to clinically relevant drugs, such as CFTR modulators. This approach is effective for the clonal expansion of single human cells and for basal cell populations from epithelial tissues from all three germ layers and therefore may be broadly applicable for modeling of epithelia. [Display omitted] •SMAD activity is active in suprabasal cells but is weaker in basal epithelial cells•SMAD signaling activity correlates with mucociliary differentiation in the airway•Dual TGFβ/BMP inhibition prevents spontaneous differentiation in culture•Dual TGFβ/BMP inhibition allows prolonged culture of diverse epithelial basal cells Mou et al. show that small-molecule-mediated SMAD signaling inhibition allows prolonged feeder-free culture of diverse functional epithelial basal stem cells in a 2D format. This methodology provides a facile patient-specific epithelial disease modeling platform, as shown by the expansion of airway epithelium from non-invasively obtained specimens from cystic fibrosis patients.

TGFβ/BMP4/SMAD signaling

p63+ basal cells

senescence

epithelial basal and stems cells

differentiation

stemness

telomeres

dedifferentiation

dual SMAD signaling inhibition

replicative exhaustion

Details

Title: Subtitle: Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells
Creators: Hongmei Mou - Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA
Vladimir Vinarsky - Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA
Purushothama Rao Tata - Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA
Karissa Brazauskas - Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA
Soon H Choi - Department of Anatomy and Cell Biology, College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Adrianne K Crooke - Department of Anatomy and Cell Biology, College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Bing Zhang - Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, 7 Divinity Avenue, Cambridge, MA 02138, USA
George M Solomon - Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35233, USA
Brett Turner - Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL 35233, USA
Hermann Bihler - CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA 01730, USA
Jan Harrington - CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA 01730, USA
Allen Lapey - Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA
Colleen Channick - Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA
Colleen Keyes - Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA
Adam Freund - Departments of Medicine and Biochemistry, Stanford University, Stanford, CA 94305, USA
Steven Artandi - Departments of Medicine and Biochemistry, Stanford University, Stanford, CA 94305, USA
Martin Mense - CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA 01730, USA
Steven Rowe - Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35233, USA
John F Engelhardt - Department of Anatomy and Cell Biology, College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Ya-Chieh Hsu - Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, 7 Divinity Avenue, Cambridge, MA 02138, USA
Jayaraj Rajagopal - Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA
Resource Type: Journal article
Publication Details: Cell stem cell, Vol.19(2), pp.217-231
Publisher: Elsevier Inc
DOI: 10.1016/j.stem.2016.05.012
PMID: 27320041
PMCID: PMC4975684
ISSN: 1934-5909
eISSN: 1875-9777
Language: English
Date published: 08/04/2016
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
Record Identifier: 9984025442602771

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