Cargando…

The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation

Basal cells (BCs) are stem/progenitor cells of the mucociliary airway epithelium, and their differentiation is orchestrated by the NOTCH signaling pathway. NOTCH3 receptor signaling regulates BC to club cell differentiation; however, the downstream responses that regulate this process are unknown. O...

Descripción completa

Detalles Bibliográficos
Autores principales: Bodas, Manish, Subramaniyan, Bharathiraja, Moore, Andrew R., Metcalf, Jordan P., Ocañas, Sarah R., Freeman, Willard M., Georgescu, Constantin, Wren, Jonathan D., Walters, Matthew S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8620267/
https://www.ncbi.nlm.nih.gov/pubmed/34831437
http://dx.doi.org/10.3390/cells10113215
_version_ 1784605179490861056
author Bodas, Manish
Subramaniyan, Bharathiraja
Moore, Andrew R.
Metcalf, Jordan P.
Ocañas, Sarah R.
Freeman, Willard M.
Georgescu, Constantin
Wren, Jonathan D.
Walters, Matthew S.
author_facet Bodas, Manish
Subramaniyan, Bharathiraja
Moore, Andrew R.
Metcalf, Jordan P.
Ocañas, Sarah R.
Freeman, Willard M.
Georgescu, Constantin
Wren, Jonathan D.
Walters, Matthew S.
author_sort Bodas, Manish
collection PubMed
description Basal cells (BCs) are stem/progenitor cells of the mucociliary airway epithelium, and their differentiation is orchestrated by the NOTCH signaling pathway. NOTCH3 receptor signaling regulates BC to club cell differentiation; however, the downstream responses that regulate this process are unknown. Overexpression of the active NOTCH3 intracellular domain (NICD3) in primary human bronchial epithelial cells (HBECs) on in vitro air–liquid interface culture promoted club cell differentiation. Bulk RNA-seq analysis identified 692 NICD3-responsive genes, including the classical NOTCH target HEYL, which increased in response to NICD3 and positively correlated with SCGB1A1 (club cell marker) expression. siRNA knockdown of HEYL decreased tight junction formation and cell proliferation. Further, HEYL knockdown reduced club, goblet and ciliated cell differentiation. In addition, we observed decreased expression of HEYL in HBECs from donors with chronic obstructive pulmonary disease (COPD) vs. normal donors which correlates with the impaired differentiation capacity of COPD cells. Finally, overexpression of HEYL in COPD HBECs promoted differentiation into club, goblet and ciliated cells, suggesting the impaired capacity of COPD cells to generate a normal airway epithelium is a reversible phenotype that can be regulated by HEYL. Overall, our data identify the NOTCH3 downstream target HEYL as a key regulator of airway epithelial differentiation.
format Online
Article
Text
id pubmed-8620267
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-86202672021-11-27 The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation Bodas, Manish Subramaniyan, Bharathiraja Moore, Andrew R. Metcalf, Jordan P. Ocañas, Sarah R. Freeman, Willard M. Georgescu, Constantin Wren, Jonathan D. Walters, Matthew S. Cells Article Basal cells (BCs) are stem/progenitor cells of the mucociliary airway epithelium, and their differentiation is orchestrated by the NOTCH signaling pathway. NOTCH3 receptor signaling regulates BC to club cell differentiation; however, the downstream responses that regulate this process are unknown. Overexpression of the active NOTCH3 intracellular domain (NICD3) in primary human bronchial epithelial cells (HBECs) on in vitro air–liquid interface culture promoted club cell differentiation. Bulk RNA-seq analysis identified 692 NICD3-responsive genes, including the classical NOTCH target HEYL, which increased in response to NICD3 and positively correlated with SCGB1A1 (club cell marker) expression. siRNA knockdown of HEYL decreased tight junction formation and cell proliferation. Further, HEYL knockdown reduced club, goblet and ciliated cell differentiation. In addition, we observed decreased expression of HEYL in HBECs from donors with chronic obstructive pulmonary disease (COPD) vs. normal donors which correlates with the impaired differentiation capacity of COPD cells. Finally, overexpression of HEYL in COPD HBECs promoted differentiation into club, goblet and ciliated cells, suggesting the impaired capacity of COPD cells to generate a normal airway epithelium is a reversible phenotype that can be regulated by HEYL. Overall, our data identify the NOTCH3 downstream target HEYL as a key regulator of airway epithelial differentiation. MDPI 2021-11-18 /pmc/articles/PMC8620267/ /pubmed/34831437 http://dx.doi.org/10.3390/cells10113215 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bodas, Manish
Subramaniyan, Bharathiraja
Moore, Andrew R.
Metcalf, Jordan P.
Ocañas, Sarah R.
Freeman, Willard M.
Georgescu, Constantin
Wren, Jonathan D.
Walters, Matthew S.
The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title_full The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title_fullStr The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title_full_unstemmed The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title_short The NOTCH3 Downstream Target HEYL Is Required for Efficient Human Airway Basal Cell Differentiation
title_sort notch3 downstream target heyl is required for efficient human airway basal cell differentiation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8620267/
https://www.ncbi.nlm.nih.gov/pubmed/34831437
http://dx.doi.org/10.3390/cells10113215
work_keys_str_mv AT bodasmanish thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT subramaniyanbharathiraja thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT mooreandrewr thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT metcalfjordanp thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT ocanassarahr thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT freemanwillardm thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT georgescuconstantin thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT wrenjonathand thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT waltersmatthews thenotch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT bodasmanish notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT subramaniyanbharathiraja notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT mooreandrewr notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT metcalfjordanp notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT ocanassarahr notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT freemanwillardm notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT georgescuconstantin notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT wrenjonathand notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation
AT waltersmatthews notch3downstreamtargetheylisrequiredforefficienthumanairwaybasalcelldifferentiation