Single-cell transcriptomics reveals e-cigarette vapor-induced airway epithelial remodeling and injury.
Humans
E-Cigarette Vapor
/ toxicity
Transcriptome
/ drug effects
Airway Remodeling
/ drug effects
Single-Cell Analysis
/ methods
Male
Cells, Cultured
Female
Middle Aged
Epithelial Cells
/ drug effects
Adult
Pulmonary Disease, Chronic Obstructive
/ pathology
Electronic Nicotine Delivery Systems
Respiratory Mucosa
/ drug effects
Bronchi
/ drug effects
Cilia
/ drug effects
Cell Differentiation
/ drug effects
COPD
Cilia
Ciliated cells
E-cigarette
Notch signaling
Journal
Respiratory research
ISSN: 1465-993X
Titre abrégé: Respir Res
Pays: England
ID NLM: 101090633
Informations de publication
Date de publication:
28 Sep 2024
28 Sep 2024
Historique:
received:
07
09
2023
accepted:
26
08
2024
medline:
29
9
2024
pubmed:
29
9
2024
entrez:
28
9
2024
Statut:
epublish
Résumé
In recent years, e-cigarettes have been used as alternatives among adult smokers. However, the impact of e-cigarette use on human bronchial epithelial (HBE) cells remains controversial. We collected primary HBE cells of healthy nonsmokers and chronic obstructive pulmonary disease (COPD) smokers, and analyzed the impact of e- cigarette vapor extract (ECE) or cigarette smoke extract (CSE) on HBE cell differentiation and injury by single-cell RNA sequencing, immunostaining, HE staining, qPCR and ELISA. We obtained serum and sputum from healthy non- smokers, smokers and e-cigarette users, and analyzed cell injury markers and mucin proteins. ECE treatment led to a distinct differentiation program of ciliated cells and unique patterns of their cell-cell communications compared with CSE. ECE treatment caused increased Notch signaling strength in a ciliated cell subpopulation, and HBE cell remodeling and injury including hypoplasia of ciliated cells and club cells, and shorter cilia. ECE-induced hypoplasia of ciliated cells and shorter cilia were ameliorated by the Notch signaling inhibition. This study reveals distinct characteristics in e-cigarette vapor-induced airway epithelial remodeling, pointing to Notch signaling pathway as a potential targeted intervention for e-cigarette vapor-caused ciliated cell differentiation defects and cilia injury. In addition, a decrease in SCGB1A1 proteins is associated with e- cigarette users, indicating a potential lung injury marker for e-cigarette users.
Sections du résumé
BACKGROUND
BACKGROUND
In recent years, e-cigarettes have been used as alternatives among adult smokers. However, the impact of e-cigarette use on human bronchial epithelial (HBE) cells remains controversial.
METHODS
METHODS
We collected primary HBE cells of healthy nonsmokers and chronic obstructive pulmonary disease (COPD) smokers, and analyzed the impact of e- cigarette vapor extract (ECE) or cigarette smoke extract (CSE) on HBE cell differentiation and injury by single-cell RNA sequencing, immunostaining, HE staining, qPCR and ELISA. We obtained serum and sputum from healthy non- smokers, smokers and e-cigarette users, and analyzed cell injury markers and mucin proteins.
RESULTS
RESULTS
ECE treatment led to a distinct differentiation program of ciliated cells and unique patterns of their cell-cell communications compared with CSE. ECE treatment caused increased Notch signaling strength in a ciliated cell subpopulation, and HBE cell remodeling and injury including hypoplasia of ciliated cells and club cells, and shorter cilia. ECE-induced hypoplasia of ciliated cells and shorter cilia were ameliorated by the Notch signaling inhibition.
CONCLUSIONS
CONCLUSIONS
This study reveals distinct characteristics in e-cigarette vapor-induced airway epithelial remodeling, pointing to Notch signaling pathway as a potential targeted intervention for e-cigarette vapor-caused ciliated cell differentiation defects and cilia injury. In addition, a decrease in SCGB1A1 proteins is associated with e- cigarette users, indicating a potential lung injury marker for e-cigarette users.
Identifiants
pubmed: 39342154
doi: 10.1186/s12931-024-02962-4
pii: 10.1186/s12931-024-02962-4
doi:
Substances chimiques
E-Cigarette Vapor
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
353Informations de copyright
© 2024. The Author(s).
Références
Reidel B, Radicioni G, Clapp PW, Ford AA, Abdelwahab S, Rebuli ME, et al. E-cigarette use causes a unique innate immune response in the lung, involving increased neutrophilic activation and altered mucin secretion. Am J Respir Crit Care Med. 2018;197:492–501.
doi: 10.1164/rccm.201708-1590OC
pubmed: 29053025
pmcid: 5821909
Ghosh A, Coakley RD, Ghio AJ, Muhlebach MS, Esther CR Jr, Alexis NE, et al. Chronic E-cigarette use increases neutrophil elastase and matrix metalloprotease levels in the lung. Am J Respir Crit Care Med. 2019;200:1392–401.
doi: 10.1164/rccm.201903-0615OC
pubmed: 31390877
pmcid: 6884043
Gellatly S, Pavelka N, Crue T, Schweitzer KS, Day BJ, Min E, et al. Nicotine-Free e-Cigarette Vapor exposure stimulates il6 and mucin production in human primary small airway epithelial cells. J Inflamm Res. 2020;13:175–85.
doi: 10.2147/JIR.S244434
pubmed: 32368126
pmcid: 7170627
Garcia-Arcos I, Geraghty P, Baumlin N, Campos M, Dabo AJ, Jundi B, et al. Chronic electronic cigarette exposure in mice induces features of COPD in a nicotine- dependent manner. Thorax. 2016;71:1119–29.
doi: 10.1136/thoraxjnl-2015-208039
pubmed: 27558745
Madison MC, Landers CT, Gu BH, Chang CY, Tung HY, You R, et al. Electronic cigarettes disrupt lung lipid homeostasis and innate immunity independent of nicotine. J Clin Invest. 2019;129:4290–304.
doi: 10.1172/JCI128531
pubmed: 31483291
pmcid: 6763255
Chung S, Baumlin N, Dennis JS, Moore R, Salathe SF, Whitney PL, et al. Electronic cigarette Vapor with nicotine causes airway mucociliary dysfunction preferentially via TRPA1 receptors. Am J Respir Crit Care Med. 2019;200:1134–45.
doi: 10.1164/rccm.201811-2087OC
pubmed: 31170808
pmcid: 6888648
Tsai M, Song MA, Mcndrew C, Brasky TM, Freudenheim JL, MathéE, et al. Electronic versus combustible cigarette effects on inflammasome component release into human lung. Am J Respir Crit Care Med. 2019;199:922–5.
doi: 10.1164/rccm.201808-1467LE
pubmed: 30608866
pmcid: 6444658
Shen Y, Wolkowicz MJ, Kotova T, Fan L, Timko MP. Transcriptome sequencing reveals e-cigarette vapor and mainstream-smoke from tobacco cigarettes activate different gene expression profiles in human bronchial epithelial cells. Sci Rep. 2016;6:23984.
doi: 10.1038/srep23984
pubmed: 27041137
pmcid: 4819171
Herr C, Tsitouras K, Niederstraßer J, Backes C, Beisswenger C, Dong L, et al. Cigarette smoke and electronic cigarettes differentially activate bronchial epithelial cells. Respir Res. 2020;21:67.
doi: 10.1186/s12931-020-1317-2
pubmed: 32164736
pmcid: 7068890
Park HR, Vallarino J, O’Sullivan M, Wirth C, Panganiban RA, Webb G, et al. Electronic cigarette smoke reduces ribosomal protein gene expression to impair protein synthesis in primary human airway epithelial cells. Sci Rep. 2021;11:17517.
doi: 10.1038/s41598-021-97013-z
pubmed: 34471210
pmcid: 8410828
Park HR, O’Sullivan M, Vallarino J, Shumyatcher M, Himes BE, Park JA, et al. Transcriptomic response of primaryhuman airway epithelial cells toflavoring chemica ls in electronic cigarettes. Sci Rep. 2019;9(1):1400.
doi: 10.1038/s41598-018-37913-9
pubmed: 30710127
pmcid: 6358614
Martin EM, Clapp PW, Rebuli ME, Pawlak EA, Glista-Baker E, Benowitz NL, et al. E-cigarette use results in suppression of immune and inflammatory-response genes in nasal epithelial cells similar to cigarette smoke. Am J Physiol Lung Cell Mol Physiol. 2016;311:L135–44.
doi: 10.1152/ajplung.00170.2016
pubmed: 27288488
pmcid: 4967187
Travaglini KJ, Nabhan AN, Penland L, Sinha R, Gillich A, Sit RV, et al. A molecular cell atlas of the human lung from single-cell RNA sequencing. Nature. 2020;587:619–25.
doi: 10.1038/s41586-020-2922-4
pubmed: 33208946
pmcid: 7704697
Goldfarbmuren KC, Jackson ND, Sajuthi SP, Dyjack N, Li KS, Rios CL, et al. Dissecting the cellular specificity of smoking effects and reconstructing lineages in the human airway epithelium. Nat Commun. 2020;11:2485.
doi: 10.1038/s41467-020-16239-z
pubmed: 32427931
pmcid: 7237663
Basnet S, Bochkov YA, Brockman-Schneider RA, Kuipers I, Aesif SW, Jackson DJ, et al. CDHR3 asthma-risk genotype affects susceptibility of airway epithelium to rhinovirus C infections. Am J Respir Cell Mol Biol. 2019;61:450–8.
doi: 10.1165/rcmb.2018-0220OC
pubmed: 30916989
pmcid: 6775945
Montoro DT, Haber AL, Biton M, Vinarsky V, Lin B, Birket SE, et al. A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature. 2018;560:319–24.
doi: 10.1038/s41586-018-0393-7
pubmed: 30069044
pmcid: 6295155
Tan FE, Vladar EK, Ma L, Fuentealba LC, Hoh R, Espinoza FH, et al. Myb promotes centriole amplification and later steps of the multiciliogenesis program. Development. 2013;140:4277–86.
doi: 10.1242/dev.094102
pubmed: 24048590
pmcid: 3787764
Vladar EK, Stratton MB, Saal ML, Salazar-De Simone G, Wang X, Wolgemuth D, et al. Cyclin-dependent kinase control of motile ciliogenesis. Elife. 2018;7:e36375.
doi: 10.7554/eLife.36375
pubmed: 30152757
pmcid: 6145839
Jin S, Guerrero-Juarez CF, Zhang L, Chang I, Ramos R, Kuan CH, et al. Inference and analysis of cell-cell communication using Cell Chat. Nat Commun. 2021;12:1088.
doi: 10.1038/s41467-021-21246-9
pubmed: 33597522
pmcid: 7889871
Rock JR, Gao X, Xue Y, Randell SH, Kong YY, Hogan BL. Notch-dependent differentiation of adult airway basal stem cells. Cell Stem Cell. 2011;8:639–48.
doi: 10.1016/j.stem.2011.04.003
pubmed: 21624809
pmcid: 3778678
Wohnhaas CT, Gindele JA, Kiechle T, Shen Y, Leparc GG, Stierstorfer B, et al. Cigarette smoke specifically affects small airway epithelial cell populations and triggers the expansion of inflammatory and squamous differentiation associated basal cells. Int J Mol Sci. 2021;22:7646.
doi: 10.3390/ijms22147646
pubmed: 34299265
pmcid: 8305830
Okuda K, Chen G, Subramani DB, Wolf M, Gilmore RC, Kato T, et al. Localization of secretory mucins MUC5AC and MUC5B in normal/healthy human airways. Am J Respir Crit Care Med. 2019;199:715–27.
doi: 10.1164/rccm.201804-0734OC
pubmed: 30352166
pmcid: 6423099
Brekman A, Walters MS, Tilley AE, Crystal RG. FOXJ1 prevents cilia growth inhibition by cigarette smoke in human airway epithelium in vitro. Am J Respir Cell Mol Biol. 2014;51:688–700.
doi: 10.1165/rcmb.2013-0363OC
pubmed: 24828273
pmcid: 4224080
Wallmeier J, Frank D, Shoemark A, Nöthe-Menchen T, Cindric S, Olbrich H, et al. De novo mutations in FOXJ1 result in a motile ciliopathy with hydrocephalus and randomization of left/right body asymmetry. Am J Hum Genet. 2019;105:1030–9.
doi: 10.1016/j.ajhg.2019.09.022
pubmed: 31630787
pmcid: 6849114
Yin W, Liontos A, Koepke J, Ghoul M, Mazzocchi L, Liu X, et al. An essential function for autocrine hedgehog signaling in epithelial proliferation and differentiation in the trachea. Development. 2022;149:dev199804.
doi: 10.1242/dev.199804
pubmed: 35112129
pmcid: 8918789
Bodas M, Subramaniyan B, Moore AR, Metcalf JP, Ocañas SR, Freeman WM, et al. The NOTCH3 downstream target HEYL is required for efficient human airway basal cell differentiation. Cells. 2021;10:3215.
doi: 10.3390/cells10113215
pubmed: 34831437
pmcid: 8620267
Bodas M, Moore AR, Subramaniyan B, Georgescu C, Wren JD, Freeman WM, et al. Cigarette smoke activates NOTCH3 to promote goblet cell differentiation in human airway epithelial cells. Am J Respir Cell Mol Biol. 2021;64:426–40.
doi: 10.1165/rcmb.2020-0302OC
pubmed: 33444514
pmcid: 8008804
Zahid M, Feinstein TN, Oro A, Schwartz M, Lee AD, Lo CW. Rapid Ex-Vivo ciliogenesis and dose-dependent effect of notch inhibition on ciliogenesis of respiratory epithelia. Biomolecules. 2020;10:1182.
doi: 10.3390/biom10081182
pubmed: 32823934
pmcid: 7464104
Bergeron C, Tulic MK, Hamid Q. Tools used to measure airway remodelling in research. Eur Respir J. 2007;29:596–604.
doi: 10.1183/09031936.00019906
pubmed: 17329494
Hogg JC, Chu F, Utokaparch S, Woods R, Elliott WM, Buzatu L, et al. The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med. 2004;350:2645–53.
doi: 10.1056/NEJMoa032158
pubmed: 15215480
Leopold PL, O’Mahony MJ, Lian XJ, Tilley AE, Harvey BG, Crystal RG. Smoking is associated with shortened airway cilia. PLoS ONE. 2009;4:e8157.
doi: 10.1371/journal.pone.0008157
pubmed: 20016779
pmcid: 2790614
Chaumont M, Bernard A, Pochet S, Mélot C, El Khattabi C, Reye F, et al. High- wattage E-cigarettes induce tissue hypoxia and lower airway injury: a randomized clinical trial. Am J Respir Crit Care Med. 2018;198:123–6.
doi: 10.1164/rccm.201711-2198LE
pubmed: 29451806
Singh KP, Lawyer G, Muthumalage T, Maremanda KP, Khan NA, McDonough SR, et al. Systemic biomarkers in electronic cigarette users: implications for noninvasive assessment of vaping-associated pulmonary injuries. ERJ Open Res. 2019;5:00182–2019.
doi: 10.1183/23120541.00182-2019
pubmed: 31886159
pmcid: 6926365
Podguski S, Kaur G, Muthumalage T, McGraw MD, Rahman I. Noninvasive systemic biomarkers of e-cigarette or vaping use-associated lung injury: a pilot study. ERJ Open Res. 2022;8:00639–2021.
doi: 10.1183/23120541.00639-2021
pubmed: 35386827
pmcid: 8977595
Belgacemi R, Luczka E, Ancel J, Diabasana Z, Perotin JM, Germain A, et al. Airway epithelial cell differentiation relies on deficient Hedgehog signalling in COPD. EBioMedicine. 2020;51:102572.
doi: 10.1016/j.ebiom.2019.11.033
pubmed: 31877414
Schmid A, Sailland J, Novak L, Baumlin N, Fregien N, Salathe M. Modulation of Wnt signaling is essential for the differentiation of ciliated epithelial cells in human airways. FEBS Lett. 2017;591:3493–506.
doi: 10.1002/1873-3468.12851
pubmed: 28921507
pmcid: 5683904
Carlier FM, Dupasquier S, Ambroise J, Detry B, Lecocq M, Biétry-Claudet C, et al. Canonical WNT pathway is activated in the airway epithelium in chronic obstructive pulmonary disease. EBioMedicine. 2020;61:103034.
doi: 10.1016/j.ebiom.2020.103034
pubmed: 33045470
pmcid: 7559244
Komura M, Sato T, Yoshikawa H, Nitta NA, Suzuki Y, Koike K, et al. Propylene glycol, a component of electronic cigarette liquid, damages epithelial cells in human small airways. Respir Res. 2022;23:216.
doi: 10.1186/s12931-022-02142-2
pubmed: 35999544
pmcid: 9400210
Bergen V, Lange M, Peidli S, Wolf FA, Theis FJ. Generalizing RNA velocity to transient cell states through dynamical modeling. Nat Biotechnol. 2020;38:1408–14.
doi: 10.1038/s41587-020-0591-3
pubmed: 32747759
Leung C, Wadsworth SJ, Yang SJ, Dorscheid DR. Structural and functional variations in human bronchial epithelial cells cultured in air-liquid interface using different growth media. Am J Physiol Lung Cell Mol Physiol. 2020;318(5):L1063–73.
doi: 10.1152/ajplung.00190.2019
pubmed: 32208929