{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T04:58:11Z","timestamp":1792645091436,"version":"4.5.0"},"reference-count":50,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2021,4,1]],"date-time":"2021-04-01T00:00:00Z","timestamp":1617235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":["www.atsjournals.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,4,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Chronic obstructive pulmonary disease (COPD) is the third leading cause of death in the United States and is primarily caused by cigarette smoking. Increased numbers of mucus-producing secretory (\u201cgoblet\u201d) cells, defined as goblet cell metaplasia or hyperplasia (GCMH), contributes significantly to COPD pathophysiology. The objective of this study was to determine whether NOTCH signaling regulates goblet cell differentiation in response to cigarette smoke. Primary human bronchial epithelial cells (HBECs) from nonsmokers and smokers with COPD were differentiated in vitro on air\u2013liquid interface and exposed to cigarette smoke extract (CSE) for 7 days. NOTCH signaling activity was modulated using 1) the NOTCH\/\u03b3-secretase inhibitor dibenzazepine (DBZ), 2) lentiviral overexpression of the NICD3 (NOTCH3-intracellular domain), or 3) NOTCH3-specific siRNA. Cell differentiation and response to CSE were evaluated by quantitative PCR, Western blotting, immunostaining, and RNA sequencing. We found that CSE exposure of nonsmoker airway epithelium induced goblet cell differentiation characteristic of GCMH. Treatment with DBZ suppressed CSE-dependent induction of goblet cell differentiation. Furthermore, CSE induced NOTCH3 activation, as revealed by increased NOTCH3 nuclear localization and elevated NICD3 protein levels. Overexpression of NICD3 increased the expression of goblet cell\u2013associated genes SPDEF and MUC5AC, whereas NOTCH3 knockdown suppressed CSE-mediated induction of SPDEF and MUC5AC. Finally, CSE exposure of COPD airway epithelium induced goblet cell differentiation in a NOTCH3-dependent manner. These results identify NOTCH3 activation as one of the important mechanisms by which cigarette smoke induces goblet cell differentiation, thus providing a novel potential strategy to control GCMH-related pathologies in smokers and patients with COPD.<\/jats:p>","DOI":"10.1165\/rcmb.2020-0302oc","type":"journal-article","created":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T18:50:57Z","timestamp":1610650257000},"page":"426-440","update-policy":"https:\/\/doi.org\/10.1164\/crossmarkpolicies","source":"Crossref","is-referenced-by-count":50,"title":["Cigarette Smoke Activates NOTCH3 to Promote Goblet Cell Differentiation in Human Airway Epithelial Cells"],"prefix":"10.1093","volume":"64","author":[{"given":"Manish","family":"Bodas","sequence":"first","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew R.","family":"Moore","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bharathiraja","family":"Subramaniyan","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Constantin","family":"Georgescu","sequence":"additional","affiliation":[{"name":"Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan D.","family":"Wren","sequence":"additional","affiliation":[{"name":"Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Willard M.","family":"Freeman","sequence":"additional","affiliation":[{"name":"Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brent R.","family":"Brown","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jordan P.","family":"Metcalf","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew S.","family":"Walters","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,4,1]]},"reference":[{"key":"2026021100470262200_bib1","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.mcna.2012.04.012","article-title":"Pathobiologic mechanisms of chronic obstructive pulmonary disease","volume":"96","author":"Tam","year":"2012","journal-title":"Med Clin North Am"},{"key":"2026021100470262200_bib2","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1186\/s12931-019-1017-y","article-title":"The pathology of small airways disease in COPD: historical aspects and future directions","volume":"20","author":"Higham","year":"2019","journal-title":"Respir Res"},{"key":"2026021100470262200_bib3","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1111\/resp.12660","article-title":"Global burden of COPD","volume":"21","author":"L\u00f3pez-Campos","year":"2016","journal-title":"Respirology"},{"key":"2026021100470262200_bib4","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3390\/ijerph6010209","article-title":"Smoking and chronic obstructive pulmonary disease (COPD). Parallel epidemics of the 21 century","volume":"6","author":"Laniado-Labor\u00edn","year":"2009","journal-title":"Int J Environ Res Public Health"},{"key":"2026021100470262200_bib5","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.stem.2014.07.012","article-title":"Repair and regeneration of the respiratory system: complexity, plasticity, and mechanisms of lung stem cell function","volume":"15","author":"Hogan","year":"2014","journal-title":"Cell Stem Cell"},{"key":"2026021100470262200_bib6","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1152\/physrev.00001.2018","article-title":"Building and regenerating the lung cell by cell","volume":"99","author":"Whitsett","year":"2019","journal-title":"Physiol Rev"},{"key":"2026021100470262200_bib7","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1146\/annurev-physiol-021014-071931","article-title":"Cilia dysfunction in lung disease","volume":"77","author":"Tilley","year":"2015","journal-title":"Annu Rev Physiol"},{"key":"2026021100470262200_bib8","doi-asserted-by":"crossref","first-page":"S210","DOI":"10.1513\/AnnalsATS.201806-439AW","article-title":"Mucociliary defense: emerging cellular, molecular, and animal models","volume":"15","author":"Benam","year":"2018","journal-title":"Ann Am Thorac Soc"},{"key":"2026021100470262200_bib9","doi-asserted-by":"crossref","first-page":"S143","DOI":"10.1513\/AnnalsATS.201802-128AW","article-title":"Airway epithelial differentiation and mucociliary clearance","volume":"15","author":"Whitsett","year":"2018","journal-title":"Ann Am Thorac Soc"},{"key":"2026021100470262200_bib10","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.pharmthera.2017.12.009","article-title":"Persistent induction of goblet cell differentiation in the airways: therapeutic approaches","volume":"185","author":"Reid","year":"2018","journal-title":"Pharmacol Ther"},{"key":"2026021100470262200_bib11","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/dgd.12628","article-title":"Notch signaling in the mammalian respiratory system, specifically the trachea and lungs, in development, homeostasis, regeneration, and disease","volume":"62","author":"Kiyokawa","year":"2020","journal-title":"Dev Growth Differ"},{"key":"2026021100470262200_bib12","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1165\/rcmb.2019-0069OC","article-title":"Blocking notch3 signaling abolishes muc5ac production in airway epithelial cells from individuals with asthma","volume":"62","author":"Reid","year":"2020","journal-title":"Am J Respir Cell Mol Biol"},{"key":"2026021100470262200_bib13","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1242\/bio.20121701","article-title":"The loss of Hoxa5 function promotes Notch-dependent goblet cell metaplasia in lung airways","volume":"1","author":"Boucherat","year":"2012","journal-title":"Biol Open"},{"key":"2026021100470262200_bib14","doi-asserted-by":"crossref","first-page":"e0116507","DOI":"10.1371\/journal.pone.0116507","article-title":"Activation of NOTCH1 or NOTCH3 signaling skews human airway basal cell differentiation toward a secretory pathway","volume":"10","author":"Gomi","year":"2015","journal-title":"PLoS One"},{"key":"2026021100470262200_bib15","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.celrep.2014.12.017","article-title":"Notch2 is required for inflammatory cytokine-driven goblet cell metaplasia in the lung","volume":"10","author":"Danahay","year":"2015","journal-title":"Cell Rep"},{"key":"2026021100470262200_bib16","doi-asserted-by":"crossref","first-page":"3533","DOI":"10.1242\/dev.063727","article-title":"Notch signaling prevents mucous metaplasia in mouse conducting airways during postnatal development","volume":"138","author":"Tsao","year":"2011","journal-title":"Development"},{"key":"2026021100470262200_bib17","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1136\/thoraxjnl-2017-210593","article-title":"NOTCH3 contributes to rhinovirus-induced goblet cell hyperplasia in COPD airway epithelial cells","volume":"74","author":"Jing","year":"2019","journal-title":"Thorax"},{"key":"2026021100470262200_bib18","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1165\/rcmb.2019-0257OC","article-title":"Antisense oligonucleotides targeting jagged 1 reduce house dust mite-induced goblet cell metaplasia in the adult murine lung","volume":"63","author":"Carrer","year":"2020","journal-title":"Am J Respir Cell Mol Biol"},{"key":"2026021100470262200_bib19","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1038\/nature15715","article-title":"Therapeutic antibodies reveal Notch control of transdifferentiation in the adult lung","volume":"528","author":"Lafkas","year":"2015","journal-title":"Nature"},{"key":"2026021100470262200_bib20","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.stem.2011.04.003","article-title":"Notch-dependent differentiation of adult airway basal stem cells","volume":"8","author":"Rock","year":"2011","journal-title":"Cell Stem Cell"},{"key":"2026021100470262200_bib21","doi-asserted-by":"crossref","first-page":"1751","DOI":"10.1242\/dev.029249","article-title":"Notch signaling promotes airway mucous metaplasia and inhibits alveolar development","volume":"136","author":"Guseh","year":"2009","journal-title":"Development"},{"key":"2026021100470262200_bib22","doi-asserted-by":"crossref","first-page":"454","DOI":"10.1007\/s12015-016-9656-6","article-title":"Jag1-mediated notch signaling regulates secretory cell differentiation of the human airway epithelium","volume":"12","author":"Gomi","year":"2016","journal-title":"Stem Cell Rev Rep"},{"key":"2026021100470262200_bib23","doi-asserted-by":"crossref","first-page":"1540","DOI":"10.1164\/rccm.201904-0792OC","article-title":"Single-cell reconstruction of human basal cell diversity in normal and idiopathic pulmonary fibrosis lung","volume":"202","author":"Carraro","year":"2020","journal-title":"Am J Respir Crit Care Med"},{"key":"2026021100470262200_bib24","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.1152\/physrev.00005.2017","article-title":"Notch signaling in development, tissue homeostasis, and disease","volume":"97","author":"Siebel","year":"2017","journal-title":"Physiol Rev"},{"key":"2026021100470262200_bib25","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1164\/rccm.201208-1449OC","article-title":"A dynamic bronchial airway gene expression signature of chronic obstructive pulmonary disease and lung function impairment","volume":"187","author":"Steiling","year":"2013","journal-title":"Am J Respir Crit Care Med"},{"key":"2026021100470262200_bib26","doi-asserted-by":"crossref","first-page":"e22798","DOI":"10.1371\/journal.pone.0022798","article-title":"Biologic phenotyping of the human small airway epithelial response to cigarette smoking","volume":"6","author":"Tilley","year":"2011","journal-title":"PLoS One"},{"key":"2026021100470262200_bib27","doi-asserted-by":"crossref","first-page":"6236","DOI":"10.4049\/jimmunol.0900614","article-title":"Regulation of airway MUC5AC expression by IL-1beta and IL-17A; the NF-kappaB paradigm","volume":"183","author":"Fujisawa","year":"2009","journal-title":"J Immunol"},{"key":"2026021100470262200_bib28","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1002\/stem.2512","article-title":"Egf-amphiregulin interplay in airway stem\/progenitor cells links the pathogenesis of smoking-induced lesions in the human airway epithelium","volume":"35","author":"Zuo","year":"2017","journal-title":"Stem Cells"},{"key":"2026021100470262200_bib29","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/1755-8794-5-21","article-title":"Genes associated with MUC5AC expression in small airway epithelium of human smokers and non-smokers","volume":"5","author":"Wang","year":"2012","journal-title":"BMC Med Genomics"},{"key":"2026021100470262200_bib30","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1111\/imm.12352","article-title":"S100A8, S100A9 and S100A12 activate airway epithelial cells to produce MUC5AC via extracellular signal-regulated kinase and nuclear factor-\u03baB pathways","volume":"144","author":"Kang","year":"2015","journal-title":"Immunology"},{"key":"2026021100470262200_bib31","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1172\/JCI29176","article-title":"SPDEF regulates goblet cell hyperplasia in the airway epithelium","volume":"117","author":"Park","year":"2007","journal-title":"J Clin Invest"},{"key":"2026021100470262200_bib32","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1038\/ncb2241","article-title":"Control of vertebrate multiciliogenesis by miR-449 through direct repression of the Delta\/Notch pathway","volume":"13","author":"Marcet","year":"2011","journal-title":"Nat Cell Biol"},{"key":"2026021100470262200_bib33","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1242\/dev.116855","article-title":"Notch3-Jagged signaling controls the pool of undifferentiated airway progenitors","volume":"142","author":"Mori","year":"2015","journal-title":"Development"},{"key":"2026021100470262200_bib34","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.stem.2014.05.009","article-title":"Dynamic changes in intracellular ROS levels regulate airway basal stem cell homeostasis through Nrf2-dependent Notch signaling","volume":"15","author":"Paul","year":"2014","journal-title":"Cell Stem Cell"},{"key":"2026021100470262200_bib35","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1002\/dvdy.23965","article-title":"Jagged1 is the major regulator of Notch-dependent cell fate in proximal airways","volume":"242","author":"Zhang","year":"2013","journal-title":"Dev Dyn"},{"key":"2026021100470262200_bib36","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.stem.2015.01.002","article-title":"Injury induces direct lineage segregation of functionally distinct airway basal stem\/progenitor cell subpopulations","volume":"16","author":"Pardo-Saganta","year":"2015","journal-title":"Cell Stem Cell"},{"key":"2026021100470262200_bib37","doi-asserted-by":"crossref","first-page":"dev177428","DOI":"10.1242\/dev.177428","article-title":"Novel dynamics of human mucociliary differentiation revealed by single-cell RNA sequencing of nasal epithelial cultures","volume":"146","author":"Ruiz Garc\u00eda","year":"2019","journal-title":"Development"},{"key":"2026021100470262200_bib38","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1164\/rccm.200705-795OC","article-title":"Down-regulation of the notch pathway in human airway epithelium in association with smoking and chronic obstructive pulmonary disease","volume":"179","author":"Tilley","year":"2009","journal-title":"Am J Respir Crit Care Med"},{"key":"2026021100470262200_bib39","doi-asserted-by":"crossref","first-page":"4726","DOI":"10.1093\/hmg\/ddt326","article-title":"Cigarette smoking induces small airway epithelial epigenetic changes with corresponding modulation of gene expression","volume":"22","author":"Buro-Auriemma","year":"2013","journal-title":"Hum Mol Genet"},{"key":"2026021100470262200_bib40","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1164\/rccm.201406-1167LE","article-title":"Airway Basal stem\/progenitor cells have diminished capacity to regenerate airway epithelium in chronic obstructive pulmonary disease","volume":"190","author":"Staudt","year":"2014","journal-title":"Am J Respir Crit Care Med"},{"key":"2026021100470262200_bib41","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1164\/rccm.201704-0667OC","article-title":"Exhaustion of airway basal progenitor cells in early and established chronic obstructive pulmonary disease","volume":"197","author":"Ghosh","year":"2018","journal-title":"Am J Respir Crit Care Med"},{"key":"2026021100470262200_bib42","doi-asserted-by":"crossref","first-page":"17963","DOI":"10.1038\/s41598-019-54292-x","article-title":"Altered generation of ciliated cells in chronic obstructive pulmonary disease","volume":"9","author":"Gohy","year":"2019","journal-title":"Sci Rep"},{"key":"2026021100470262200_bib43","doi-asserted-by":"crossref","first-page":"e50487","DOI":"10.7554\/eLife.50487","article-title":"Jagged and Delta-like ligands control distinct events during airway progenitor cell differentiation","volume":"8","author":"Stupnikov","year":"2019","journal-title":"Elife"},{"key":"2026021100470262200_bib44","doi-asserted-by":"crossref","first-page":"641","DOI":"10.3892\/ol.2015.3329","article-title":"Cigarette smoke induces the expression of Notch3, not Notch1, protein in lung adenocarcinoma","volume":"10","author":"Cheng","year":"2015","journal-title":"Oncol Lett"},{"key":"2026021100470262200_bib45","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1007\/s00011-015-0816-y","article-title":"Different profiles of notch signaling in cigarette smoke-induced pulmonary emphysema and bleomycin-induced pulmonary fibrosis","volume":"64","author":"Li","year":"2015","journal-title":"Inflamm Res"},{"key":"2026021100470262200_bib46","doi-asserted-by":"crossref","first-page":"e1004751","DOI":"10.1371\/journal.pgen.1004751","article-title":"Notch3 interactome analysis identified WWP2 as a negative regulator of Notch3 signaling in ovarian cancer","volume":"10","author":"Jung","year":"2014","journal-title":"PLoS Genet"},{"key":"2026021100470262200_bib47","doi-asserted-by":"crossref","first-page":"2594","DOI":"10.1016\/j.biocel.2009.08.019","article-title":"Lysosome-dependent degradation of Notch3","volume":"41","author":"Jia","year":"2009","journal-title":"Int J Biochem Cell Biol"},{"key":"2026021100470262200_bib48","doi-asserted-by":"crossref","first-page":"30855","DOI":"10.18632\/oncotarget.8806","article-title":"N-acetylcysteine negatively regulates Notch3 and its malignant signaling","volume":"7","author":"Zhang","year":"2016","journal-title":"Oncotarget"},{"key":"2026021100470262200_bib49","doi-asserted-by":"crossref","first-page":"101544","DOI":"10.1016\/j.redox.2020.101544","article-title":"Oxidative stress-based therapeutics in COPD","volume":"33","author":"Barnes","year":"2020","journal-title":"Redox Biol"},{"key":"2026021100470262200_bib50","doi-asserted-by":"crossref","first-page":"C73","DOI":"10.1152\/ajpcell.00110.2016","article-title":"Cigarette smoke-induced autophagy impairment accelerates lung aging, COPD-emphysema exacerbations and pathogenesis","volume":"314","author":"Vij","year":"2018","journal-title":"Am J Physiol Cell Physiol"}],"container-title":["American Journal of Respiratory Cell and Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/ajrcmb\/article-pdf\/64\/4\/426\/66766135\/ajrcmb_64_4_426.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/ajrcmb\/article-pdf\/64\/4\/426\/66766135\/ajrcmb_64_4_426.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T05:47:15Z","timestamp":1770788835000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/ajrcmb\/article\/64\/4\/426\/8461126"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,1]]},"references-count":50,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4,1]]},"published-print":{"date-parts":[[2021,4,1]]}},"URL":"https:\/\/doi.org\/10.1165\/rcmb.2020-0302oc","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.07.09.195818","asserted-by":"object"}]},"ISSN":["1044-1549","1535-4989"],"issn-type":[{"value":"1044-1549","type":"print"},{"value":"1535-4989","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2021,4]]},"published":{"date-parts":[[2021,4,1]]},"assertion":[{"value":"2020-07-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-01-14","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-04-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}