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Genes coding for transcription factors involved in stem cell maintenance are repressed by TGF-β and downstream of Slug/Snail2 in COPD bronchial epithelial progenitors

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Abstract

Background

Basal stem/progenitor cells of airway epithelium from chronic obstructive pulmonary disease (COPD) patients have a decrease in differentiation and self-renewal potential. Our study aimed at identifying deregulations in the genetic program of these cells that could account for their exhaustion, focusing on genes downstream of the epithelial–mesenchymal transition-inducing transcription factor Slug/Snail2 and responding to transforming growth factor (TGF)-β. TGF-β is at higher levels in COPD patient lungs, plays a role in stem/progenitor cell fate and regulates the expression of Slug/Snail2 that is highly expressed in airway basal stem/progenitors.

Methods and results

We reanalyzed a gene expression dataset that we generated from COPD and normal primary bronchial basal progenitor cells knocked down for Slug/Snail2 gene. Among the genes that we identified to be repressed downstream of Slug/Snail2 in COPD, we selected those responding to differentiation and TGF-β. The large majority of these genes are upregulated with differentiation but repressed by TGF-β. Pathway and ontology enrichment analysis revealed a set of genes coding for transcription factors involved in stem cell maintenance that are repressed downstream of Slug/Snail2 and by TGF-β in COPD but not normal basal progenitor cells. We also reveal a link between Slug/Snail2 expression and the repressive effect of TGF-β on these stem cell maintenance genes.

Conclusion

Our work brings a new insight and molecular perspective to the exhaustion of basal stem/progenitor cells observed in the airway epithelium of COPD patients, revealing that stem cell maintenance genes are repressed in these cells, with TGF-β and Slug/Snail2 being involved in this deregulation.

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Data availability

Data analyzed in this study are deposited in the NCBI’s Gene Expression Omnibus (GEO) database (GSE123129 and GSE122957).

References

  1. Rigden HM, Alias A, Havelock T, O’Donnell R, Djukanovic R, Davies DE, Wilson SJ (2016) Squamous metaplasia is increased in the bronchial epithelium of smokers with chronic obstructive pulmonary disease. PLoS ONE 11(5):e0156009

    Article  Google Scholar 

  2. Sohal SS, Walters EH (2013) Role of epithelial mesenchymal transition (EMT) in chronic obstructive pulmonary disease (COPD). Respir Res 14:120

    Article  Google Scholar 

  3. Rock JR, Randell SH, Hogan BL (2010) Airway basal stem cells: a perspective on their roles in epithelial homeostasis and remodeling. Dis Model Mech 3(9–10):545–556

    Article  CAS  Google Scholar 

  4. Steiling K, van den Berge M, Hijazi K, Florido R, Campbell J, Liu G, Xiao J, Zhang X, Duclos G, Drizik E, Si H, Perdomo C, Dumont C, Coxson HO, Alekseyev YO, Sin D, Pare P, Hogg JC, McWilliams A, Hiemstra PS, Sterk PJ, Timens W, Chang JT, Sebastiani P, O’Connor GT, Bild AH, Postma DS, Lam S, Spira A, Lenburg ME (2013) A dynamic bronchial airway gene expression signature of chronic obstructive pulmonary disease and lung function impairment. Am J Respir Crit Care Med 187(9):933–942

    Article  CAS  Google Scholar 

  5. Ghosh M, Miller YE, Nakachi I, Kwon JB, Baron AE, Brantley AE, Merrick DT, Franklin WA, Keith RL, Vandivier RW (2018) Exhaustion of airway basal progenitor cells in early and established chronic obstructive pulmonary disease. Am J Respir Crit Care Med 197(7):885–896

    Article  CAS  Google Scholar 

  6. Staudt MR, Buro-Auriemma LJ, Walters MS, Salit J, Vincent T, Shaykhiev R, Mezey JG, Tilley AE, Kaner RJ, Ho MW, Crystal RG (2014) Airway Basal stem/progenitor cells have diminished capacity to regenerate airway epithelium in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 190(8):955–958

    Article  Google Scholar 

  7. Baraldo S, Turato G, Saetta M (2012) Pathophysiology of the small airways in chronic obstructive pulmonary disease. Respiration 84(2):89–97

    Article  Google Scholar 

  8. Hogg J (2004) Peripheral lung remodelling in asthma and chronic obstructive pulmonary disease. Eur Respir J 24(6):893–894

    Article  CAS  Google Scholar 

  9. Gohy ST, Hupin C, Fregimilicka C, Detry BR, Bouzin C, Gaide Chevronay H, Lecocq M, Weynand B, Ladjemi MZ, Pierreux CE, Birembaut P, Polette M, Pilette C (2015) Imprinting of the COPD airway epithelium for dedifferentiation and mesenchymal transition. Eur Respir J 45(5):1258–1272

    Article  Google Scholar 

  10. Mertens TCJ, Karmouty-Quintana H, Taube C, Hiemstra PS (2017) Use of airway epithelial cell culture to unravel the pathogenesis and study treatment in obstructive airway diseases. Pulm Pharmacol Ther 45:101–113

    Article  CAS  Google Scholar 

  11. Nieto MA, Huang RY, Jackson RA, Thiery JP (2016) Emt: 2016. Cell 166(1):21–45

    Article  CAS  Google Scholar 

  12. Hackett NR, Shaykhiev R, Walters MS, Wang R, Zwick RK, Ferris B, Witover B, Salit J, Crystal RG (2011) The human airway epithelial basal cell transcriptome. PLoS ONE 6(5):e18378

    Article  CAS  Google Scholar 

  13. Rock JR, Onaitis MW, Rawlins EL, Lu Y, Clark CP, Xue Y, Randell SH, Hogan BL (2009) Basal cells as stem cells of the mouse trachea and human airway epithelium. Proc Natl Acad Sci USA 106(31):12771–12775

    Article  CAS  Google Scholar 

  14. Ben Brahim C, Courageux C, Jolly A, Ouine B, Cartier A, de la Grange P, de Koning L, Leroy P (2021) Proliferation genes repressed by TGF-beta are downstream of Slug/Snail2 in normal bronchial epithelial progenitors and are deregulated in COPD. Stem Cell Rev Rep. https://doi.org/10.1007/s12015-021-10123-z

    Article  PubMed  Google Scholar 

  15. Mahmood MQ, Reid D, Ward C, Muller HK, Knight DA, Sohal SS, Walters EH (2017) Transforming growth factor (TGF) beta1 and Smad signalling pathways: a likely key to EMT-associated COPD pathogenesis. Respirology 22(1):133–140

    Article  Google Scholar 

  16. Slabakova E, Pernicova Z, Slavickova E, Starsichova A, Kozubik A, Soucek K (2011) TGF-beta1-induced EMT of non-transformed prostate hyperplasia cells is characterized by early induction of SNAI2/Slug. Prostate 71(12):1332–1343

    Article  CAS  Google Scholar 

  17. Yumoto K, Thomas PS, Lane J, Matsuzaki K, Inagaki M, Ninomiya-Tsuji J, Scott GJ, Ray MK, Ishii M, Maxson R, Mishina Y, Kaartinen V (2013) TGF-beta-activated kinase 1 (Tak1) mediates agonist-induced Smad activation and linker region phosphorylation in embryonic craniofacial neural crest-derived cells. J Biol Chem 288(19):13467–13480

    Article  CAS  Google Scholar 

  18. Haw R, Stein L (2012) Using the reactome database. Curr Protoc Bioinform. https://doi.org/10.1002/0471250953.bi0807s38

    Article  Google Scholar 

  19. da Huang W, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4(1):44–57

    Article  CAS  Google Scholar 

  20. Kanehisa M, Goto S, Sato Y, Furumichi M, Tanabe M (2012) KEGG for integration and interpretation of large-scale molecular data sets. Nucleic Acids Res 40(Database issue):D109-114

    Article  CAS  Google Scholar 

  21. Leroy P, Mostov KE (2007) Slug is required for cell survival during partial epithelial–mesenchymal transition of HGF-induced tubulogenesis. Mol Biol Cell 18(5):1943–1952

    Article  CAS  Google Scholar 

  22. Cardoso WV, Williams MC, Mitsialis SA, Joyce-Brady M, Rishi AK, Brody JS (1995) Retinoic acid induces changes in the pattern of airway branching and alters epithelial cell differentiation in the developing lung in vitro. Am J Respir Cell Mol Biol 12(5):464–476

    Article  CAS  Google Scholar 

  23. Oshima T, Gedda K, Koseki J, Chen X, Husmark J, Watari J, Miwa H, Pierrou S (2011) Establishment of esophageal-like non-keratinized stratified epithelium using normal human bronchial epithelial cells. Am J Physiol Cell Physiol 300(6):C1422-1429

    Article  CAS  Google Scholar 

  24. Liu Z, Li W, Lv J, Xie R, Huang H, Li Y, He Y, Jiang J, Chen B, Guo S, Chen L (2016) Identification of potential COPD genes based on multi-omics data at the functional level. Mol Biosyst 12(1):191–204

    Article  CAS  Google Scholar 

  25. Metzger DE, Stahlman MT, Shannon JM (2008) Misexpression of ELF5 disrupts lung branching and inhibits epithelial differentiation. Dev Biol 320(1):149–160

    Article  CAS  Google Scholar 

  26. Que J, Luo X, Schwartz RJ, Hogan BL (2009) Multiple roles for SoX2 in the developing and adult mouse trachea. Development 136(11):1899–1907

    Article  CAS  Google Scholar 

  27. Tompkins DH, Besnard V, Lange AW, Wert SE, Keiser AR, Smith AN, Lang R, Whitsett JA (2009) SoX2 is required for maintenance and differentiation of bronchiolar Clara, ciliated, and goblet cells. PLoS ONE 4(12):e8248

    Article  Google Scholar 

  28. Wang J, Dai N, Cheng DH, Gao JY, Zheng JQ, Liu ZG (2020) STAT3 gene polymorphism in chronic obstructive pulmonary disease. Eur Rev Med Pharmacol Sci 24(18):9618–9625

    CAS  PubMed  Google Scholar 

  29. Yoo S, Takikawa S, Geraghty P, Argmann C, Campbell J, Lin L, Huang T, Tu Z, Foronjy RF, Spira A, Schadt EE, Powell CA, Zhu J (2015) Integrative analysis of DNA methylation and gene expression data identifies EPAS1 as a key regulator of COPD. PLoS Genet 11(1):e1004898

    Article  Google Scholar 

  30. Geismann C, Arlt A, Bauer I, Pfeifer M, Schirmer U, Altevogt P, Muerkoster SS, Schafer H (2011) Binding of the transcription factor Slug to the L1CAM promoter is essential for transforming growth factor-beta1 (TGF-beta)-induced L1CAM expression in human pancreatic ductal adenocarcinoma cells. Int J Oncol 38(1):257–266

    CAS  PubMed  Google Scholar 

  31. Joseph MJ, Dangi-Garimella S, Shields MA, Diamond ME, Sun L, Koblinski JE, Munshi HG (2009) Slug is a downstream mediator of transforming growth factor-beta1-induced matrix metalloproteinase-9 expression and invasion of oral cancer cells. J Cell Biochem 108(3):726–736

    Article  CAS  Google Scholar 

  32. Gohy ST, Detry BR, Lecocq M, Bouzin C, Weynand BA, Amatngalim GD, Sibille YM, Pilette C (2014) Polymeric immunoglobulin receptor down-regulation in chronic obstructive pulmonary disease. Persistence in the cultured epithelium and role of transforming growth factor-beta. Am J Respir Crit Care Med 190(5):509–521

    Article  CAS  Google Scholar 

  33. Gohy S, Carlier FM, Fregimilicka C, Detry B, Lecocq M, Ladjemi MZ, Verleden S, Hoton D, Weynand B, Bouzin C, Pilette C (2019) Altered generation of ciliated cells in chronic obstructive pulmonary disease. Sci Rep 9(1):17963

    Article  Google Scholar 

  34. Godinas L, Corhay JL, Henket M, Guiot J, Louis R, Moermans C (2017) Increased production of TGF-beta1 from sputum cells of COPD: relationship with airway obstruction. Cytokine 99:1–8

    Article  CAS  Google Scholar 

  35. Stoll P, Wuertemberger U, Bratke K, Zingler C, Virchow JC, Lommatzsch M (2012) Stage-dependent association of BDNF and TGF-beta1 with lung function in stable COPD. Respir Res 13:116

    Article  CAS  Google Scholar 

  36. Zhang S, Xie JG, Su BT, Li JL, Hu N, Chen J, Luo GW, Cui TP (2015) MFG-E8, a clearance glycoprotein of apoptotic cells, as a new marker of disease severity in chronic obstructive pulmonary disease. Braz J Med Biol Res 48(11):1032–1038

    Article  CAS  Google Scholar 

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Acknowledgements

We thank the Thoracic Surgery Department, the Pathology Department and the Pulmonary Department at the University Hospital Bichat-Claude Bernard (Paris, France) and the INSERM UMR1152 for providing lung tissues and for isolating the cells.

Funding

PL is supported by the French National Center for Scientific Research (CNRS). This work was funded by a donation of the Association Science et Technologie (Groupe Servier) (140065/ZA) to PL and by funding from the French National Institute for Medical Research (INSERM).

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Authors and Affiliations

  1. Inserm UMR1152, Physiopathology and Epidemiology of Respiratory Diseases, Paris, France

    Charlotte Courageux, Chamseddine Ben Brahim & Pascale Leroy

  2. Faculty of Medicine, Paris Diderot University, Bichat Campus, Paris, France

    Charlotte Courageux, Chamseddine Ben Brahim & Pascale Leroy

  3. GenoSplice, Paris, France

    Pierre de la Grange & Ariane Jolly

Authors
  1. Pierre de la Grange
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  4. Chamseddine Ben Brahim
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  5. Pascale Leroy
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Contributions

Study conception and design: PL; Conceived and Designed Experiments PL, PdlG; Conducted experiments, analyzed and interpreted data: CBB, CC, AJ, PdlG, PL; Writing—draft preparation, review and editing: PL, PdlG; Supervision: PL, PdlG; Funding acquisition: PL.

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Correspondence to Pascale Leroy.

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The authors declare that they have no conflict of interest.

Ethical approval

This study was done in accordance to the principles of the Declaration of Helsinki. Approval IRB 00006477 was obtained from Paris 7 University ethics committee, Paris Nord, France.

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Subjects included in this study gave their informed consent.

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de la Grange, P., Jolly, A., Courageux, C. et al. Genes coding for transcription factors involved in stem cell maintenance are repressed by TGF-β and downstream of Slug/Snail2 in COPD bronchial epithelial progenitors. Mol Biol Rep 48, 6729–6738 (2021). https://doi.org/10.1007/s11033-021-06664-8

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  • DOI: https://doi.org/10.1007/s11033-021-06664-8

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