Identification of human host factors required for beta-defensin-2 expression in intestinal epithelial cells upon a bacterial challenge.
Antimicrobial peptide
Bacterial challenge
Beta-defensin-2
Gene regulation
Human intestinal epithelial cell
Innate immunity
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
04 Jul 2024
04 Jul 2024
Historique:
received:
07
03
2024
accepted:
01
07
2024
medline:
5
7
2024
pubmed:
5
7
2024
entrez:
4
7
2024
Statut:
epublish
Résumé
The human intestinal tract is colonized with microorganisms, which present a diverse array of immunological challenges. A number of antimicrobial mechanisms have evolved to cope with these challenges. A key defense mechanism is the expression of inducible antimicrobial peptides (AMPs), such as beta-defensins, which rapidly inactivate microorganisms. We currently have a limited knowledge of mechanisms regulating the inducible expression of AMP genes, especially factors from the host required in these regulatory mechanisms. To identify the host factors required for expression of the beta-defensin-2 gene (HBD2) in intestinal epithelial cells upon a bacterial challenge, we performed a RNAi screen using a siRNA library spanning the whole human genome. The screening was performed in duplicate to select the strongest 79 and 110 hit genes whose silencing promoted or inhibited HBD2 expression, respectively. A set of 57 hits selected among the two groups of genes was subjected to a counter-screening and a subset was subsequently validated for its impact onto HBD2 expression. Among the 57 confirmed hits, we brought out the TLR5-MYD88 signaling pathway, but above all new signaling proteins, epigenetic regulators and transcription factors so far unrevealed in the HBD2 regulatory circuits, like the GATA6 transcription factor involved in inflammatory bowel diseases. This study represents a significant step toward unveiling the key molecular requirements to promote AMP expression in human intestinal epithelial cells, and revealing new potential targets for the development of an innovative therapeutic strategy aiming at stimulating the host AMP expression, at the era of antimicrobial resistance.
Identifiants
pubmed: 38965312
doi: 10.1038/s41598-024-66568-y
pii: 10.1038/s41598-024-66568-y
doi:
Substances chimiques
beta-Defensins
0
DEFB4A protein, human
0
RNA, Small Interfering
0
Myeloid Differentiation Factor 88
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
15442Informations de copyright
© 2024. The Author(s).
Références
Hoffmann, J. A., Kafatos, F. C., Janeway, C. A. & Ezekowitz, R. A. Phylogenetic perspectives in innate immunity. Science 284, 1313–1318. https://doi.org/10.1126/science.284.5418.1313 (1999).
doi: 10.1126/science.284.5418.1313
pubmed: 10334979
Janeway, C. A. Jr. & Medzhitov, R. Innate immune recognition. Annu. Rev. Immunol. 20, 197–216. https://doi.org/10.1146/annurev.immunol.20.083001.084359 (2002).
doi: 10.1146/annurev.immunol.20.083001.084359
pubmed: 11861602
Ganz, T. Defensins: Antimicrobial peptides of innate immunity. Nat. Rev. Immunol. 3, 710–720. https://doi.org/10.1038/nri1180 (2003).
doi: 10.1038/nri1180
pubmed: 12949495
Semple, F. & Dorin, J. R. beta-Defensins: Multifunctional modulators of infection, inflammation and more?. J. Innate Immun. 4, 337–348. https://doi.org/10.1159/000336619 (2012).
doi: 10.1159/000336619
pubmed: 22441423
pmcid: 6784047
Lehrer, R. I. et al. Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. J. Clin. Invest. 84, 553–561. https://doi.org/10.1172/JCI114198 (1989).
doi: 10.1172/JCI114198
pubmed: 2668334
pmcid: 548915
Brogden, K. A. Antimicrobial peptides: Pore formers or metabolic inhibitors in bacteria?. Nat. Rev. Microbiol. 3, 238–250. https://doi.org/10.1038/nrmicro1098 (2005).
doi: 10.1038/nrmicro1098
pubmed: 15703760
Yang, D., Biragyn, A., Hoover, D. M., Lubkowski, J. & Oppenheim, J. J. Multiple roles of antimicrobial defensins, cathelicidins, and eosinophil-derived neurotoxin in host defense. Annu. Rev. Immunol. 22, 181–215. https://doi.org/10.1146/annurev.immunol.22.012703.104603 (2004).
doi: 10.1146/annurev.immunol.22.012703.104603
pubmed: 15032578
Yang, D. et al. Beta-defensins: Linking innate and adaptive immunity through dendritic and T cell CCR6. Science 286, 525–528. https://doi.org/10.1126/science.286.5439.525 (1999).
doi: 10.1126/science.286.5439.525
pubmed: 10521347
Fellermann, K., Wehkamp, J., Herrlinger, K. R. & Stange, E. F. Crohn’s disease: A defensin deficiency syndrome?. Eur. J. Gastroenterol. Hepatol. 15, 627–634. https://doi.org/10.1097/00042737-200306000-00008 (2003).
doi: 10.1097/00042737-200306000-00008
pubmed: 12840673
Rivas-Santiago, B., Serrano, C. J. & Enciso-Moreno, J. A. Susceptibility to infectious diseases based on antimicrobial peptide production. Infect. Immun. 77, 4690–4695. https://doi.org/10.1128/IAI.01515-08 (2009).
doi: 10.1128/IAI.01515-08
pubmed: 19703980
pmcid: 2772553
Sperandio, B. et al. Virulent Shigella flexneri subverts the host innate immune response through manipulation of antimicrobial peptide gene expression. J. Exp. Med. 205, 1121–1132. https://doi.org/10.1084/jem.20071698 (2008).
doi: 10.1084/jem.20071698
pubmed: 18426984
pmcid: 2373844
MacRedmond, R., Greene, C., Taggart, C. C., McElvaney, N. & O’Neill, S. Respiratory epithelial cells require toll-like receptor 4 for induction of human beta-defensin 2 by lipopolysaccharide. Respir. Res. 6, 116. https://doi.org/10.1186/1465-9921-6-116 (2005).
doi: 10.1186/1465-9921-6-116
pubmed: 16219107
pmcid: 1276817
Ogushi, K. et al. Gangliosides act as co-receptors for Salmonella enteritidis FliC and promote FliC induction of human beta-defensin-2 expression in Caco-2 cells. J. Biol. Chem. 279, 12213–12219. https://doi.org/10.1074/jbc.M307944200 (2004).
doi: 10.1074/jbc.M307944200
pubmed: 14707135
Salzman, N. H. et al. Enteric defensins are essential regulators of intestinal microbial ecology. Nat. Immunol. 11, 76–83. https://doi.org/10.1038/ni.1825 (2010).
doi: 10.1038/ni.1825
pubmed: 19855381
Cieslik, M., Baginska, N., Gorski, A. & Jonczyk-Matysiak, E. Human beta-defensin 2 and its postulated role in modulation of the immune response. Cells 10, 2991. https://doi.org/10.3390/cells10112991 (2021).
doi: 10.3390/cells10112991
pubmed: 34831214
pmcid: 8616480
Fu, J. et al. Mechanisms and regulation of defensins in host defense. Signal Transduct. Target. Ther. 8, 300. https://doi.org/10.1038/s41392-023-01553-x (2023).
doi: 10.1038/s41392-023-01553-x
pubmed: 37574471
pmcid: 10423725
Boughan, P. K. et al. Nucleotide-binding oligomerization domain-1 and epidermal growth factor receptor: Critical regulators of beta-defensins during Helicobacter pylori infection. J. Biol. Chem. 281, 11637–11648. https://doi.org/10.1074/jbc.M510275200 (2006).
doi: 10.1074/jbc.M510275200
pubmed: 16513653
Kumar, A., Yin, J., Zhang, J. & Yu, F. S. Modulation of corneal epithelial innate immune response to pseudomonas infection by flagellin pretreatment. Invest. Ophthalmol. Vis. Sci. 48, 4664–4670. https://doi.org/10.1167/iovs.07-0473 (2007).
doi: 10.1167/iovs.07-0473
pubmed: 17898290
Tsutsumi-Ishii, Y. & Nagaoka, I. Modulation of human beta-defensin-2 transcription in pulmonary epithelial cells by lipopolysaccharide-stimulated mononuclear phagocytes via proinflammatory cytokine production. J. Immunol. 170, 4226–4236. https://doi.org/10.4049/jimmunol.170.8.4226 (2003).
doi: 10.4049/jimmunol.170.8.4226
pubmed: 12682256
Voss, E. et al. NOD2/CARD15 mediates induction of the antimicrobial peptide human beta-defensin-2. J. Biol. Chem. 281, 2005–2011. https://doi.org/10.1074/jbc.M511044200 (2006).
doi: 10.1074/jbc.M511044200
pubmed: 16319062
Wang, X. et al. Airway epithelia regulate expression of human beta-defensin 2 through Toll-like receptor 2. FASEB J. 17, 1727–1729. https://doi.org/10.1096/fj.02-0616fje (2003).
doi: 10.1096/fj.02-0616fje
pubmed: 12958190
Krisanaprakornkit, S., Kimball, J. R. & Dale, B. A. Regulation of human beta-defensin-2 in gingival epithelial cells: The involvement of mitogen-activated protein kinase pathways, but not the NF-kappaB transcription factor family. J. Immunol. 168, 316–324. https://doi.org/10.4049/jimmunol.168.1.316 (2002).
doi: 10.4049/jimmunol.168.1.316
pubmed: 11751976
Moon, S. K. et al. Activation of a Src-dependent Raf-MEK1/2-ERK signaling pathway is required for IL-1alpha-induced upregulation of beta-defensin 2 in human middle ear epithelial cells. Biochim. Biophys. Acta. 1590, 41–51. https://doi.org/10.1016/s0167-4889(02)00196-9 (2002).
doi: 10.1016/s0167-4889(02)00196-9
pubmed: 12063167
Jang, B. C. et al. Up-regulation of human beta-defensin 2 by interleukin-1beta in A549 cells: Involvement of PI3K, PKC, p38 MAPK, JNK, and NF-kappaB. Biochem. Biophys. Res. Commun. 320, 1026–1033. https://doi.org/10.1016/j.bbrc.2004.06.049 (2004).
doi: 10.1016/j.bbrc.2004.06.049
pubmed: 15240151
Kao, C. Y. et al. IL-17 markedly up-regulates beta-defensin-2 expression in human airway epithelium via JAK and NF-kappaB signaling pathways. J. Immunol. 173, 3482–3491. https://doi.org/10.4049/jimmunol.173.5.3482 (2004).
doi: 10.4049/jimmunol.173.5.3482
pubmed: 15322213
Tokunaga, F. et al. Specific recognition of linear polyubiquitin by A20 zinc finger 7 is involved in NF-kappaB regulation. EMBO J. 31, 3856–3870. https://doi.org/10.1038/emboj.2012.241 (2012).
doi: 10.1038/emboj.2012.241
pubmed: 23032187
pmcid: 3463848
Han, J., Wu, J. & Silke, J. An overview of mammalian p38 mitogen-activated protein kinases, central regulators of cell stress and receptor signaling. F1000Res 9, 653. https://doi.org/10.12688/f1000research.22092.1 (2020).
doi: 10.12688/f1000research.22092.1
Jalouli, M., Mokas, S., Turgeon, C. A., Lamalice, L. & Richard, D. E. Selective HIF-1 regulation under nonhypoxic conditions by the p42/p44 MAP kinase inhibitor PD184161. Mol. Pharmacol. 92, 510–518. https://doi.org/10.1124/mol.117.108654 (2017).
doi: 10.1124/mol.117.108654
pubmed: 28814529
Vance, R. E. The NAIP/NLRC4 inflammasomes. Curr. Opin. Immunol. 32, 84–89. https://doi.org/10.1016/j.coi.2015.01.010 (2015).
doi: 10.1016/j.coi.2015.01.010
pubmed: 25621709
Zheng, D., Kern, L. & Elinav, E. The NLRP6 inflammasome. Immunology 162, 281–289. https://doi.org/10.1111/imm.13293 (2021).
doi: 10.1111/imm.13293
pubmed: 33314083
Rodriguez, N. J., Gupta, N. & Gibson, J. Autoimmune enteropathy in an ulcerative colitis patient. ACG Case Rep. J. 5, e78. https://doi.org/10.14309/crj.2018.78 (2018).
doi: 10.14309/crj.2018.78
pubmed: 30465008
pmcid: 6224868
Steubesand, N. et al. The expression of the beta-defensins hBD-2 and hBD-3 is differentially regulated by NF-kappaB and MAPK/AP-1 pathways in an in vitro model of Candida esophagitis. BMC Immunol. 10, 36. https://doi.org/10.1186/1471-2172-10-36 (2009).
doi: 10.1186/1471-2172-10-36
pubmed: 19523197
pmcid: 2702365
Tomita, T. et al. Molecular mechanisms underlying human beta-defensin-2 gene expression in a human airway cell line (LC2/ad). Respirology 7, 305–310. https://doi.org/10.1046/j.1440-1843.2002.00415.x (2002).
doi: 10.1046/j.1440-1843.2002.00415.x
pubmed: 12421237
Koeninger, L. et al. Human beta-defensin 2 mediated immune modulation as treatment for experimental colitis. Front. Immunol. 11, 93. https://doi.org/10.3389/fimmu.2020.00093 (2020).
doi: 10.3389/fimmu.2020.00093
pubmed: 32076420
pmcid: 7006816
Aldhous, M. C., Noble, C. L. & Satsangi, J. Dysregulation of human beta-defensin-2 protein in inflammatory bowel disease. PLoS One 4, e6285. https://doi.org/10.1371/journal.pone.0006285 (2009).
doi: 10.1371/journal.pone.0006285
pubmed: 19617917
pmcid: 2708916
Laudisi, F. et al. GATA6 deficiency leads to epithelial barrier dysfunction and enhances susceptibility to gut inflammation. J. Crohns Colitis 16, 301–311. https://doi.org/10.1093/ecco-jcc/jjab145 (2022).
doi: 10.1093/ecco-jcc/jjab145
pubmed: 34374415
Kamino, Y. et al. HBD-2 is downregulated in oral carcinoma cells by DNA hypermethylation, and increased expression of hBD-2 by DNA demethylation and gene transfection inhibits cell proliferation and invasion. Oncol. Rep. 32, 462–468. https://doi.org/10.3892/or.2014.3260 (2014).
doi: 10.3892/or.2014.3260
pubmed: 24927104
pmcid: 4091880
Yin, L. & Chung, W. O. Epigenetic regulation of human beta-defensin 2 and CC chemokine ligand 20 expression in gingival epithelial cells in response to oral bacteria. Mucosal. Immunol. 4, 409–419. https://doi.org/10.1038/mi.2010.83 (2011).
doi: 10.1038/mi.2010.83
pubmed: 21248725
pmcid: 3118861
Fischer, N. et al. Histone deacetylase inhibition enhances antimicrobial peptide but not inflammatory cytokine expression upon bacterial challenge. Proc. Natl. Acad. Sci. U S A 113, E2993–E3001. https://doi.org/10.1073/pnas.1605997113 (2016).
doi: 10.1073/pnas.1605997113
pubmed: 27162363
pmcid: 4889416
Gschwandtner, M. et al. Fetal human keratinocytes produce large amounts of antimicrobial peptides: Involvement of histone-methylation processes. J. Invest. Dermatol. 134, 2192–2201. https://doi.org/10.1038/jid.2014.165 (2014).
doi: 10.1038/jid.2014.165
pubmed: 24694903
pmcid: 4447892
Liu, W. et al. Brd4 and JMJD6-associated anti-pause enhancers in regulation of transcriptional pause release. Cell 155, 1581–1595. https://doi.org/10.1016/j.cell.2013.10.056 (2013).
doi: 10.1016/j.cell.2013.10.056
pubmed: 24360279
pmcid: 3886918
Alland, L. et al. Identification of mammalian Sds3 as an integral component of the Sin3/histone deacetylase corepressor complex. Mol. Cell. Biol. 22, 2743–2750. https://doi.org/10.1128/MCB.22.8.2743-2750.2002 (2002).
doi: 10.1128/MCB.22.8.2743-2750.2002
pubmed: 11909966
pmcid: 133736
Zhang, K., Dai, X., Wallingford, M. C. & Mager, J. Depletion of Suds3 reveals an essential role in early lineage specification. Dev. Biol. 373, 359–372. https://doi.org/10.1016/j.ydbio.2012.10.026 (2013).
doi: 10.1016/j.ydbio.2012.10.026
pubmed: 23123966
Reddy-Vari, H., Kim, Y., Rajput, C. & Sajjan, U. S. Increased expression of miR146a dysregulates TLR2-induced HBD2 in airway epithelial cells from patients with COPD. ERJ Open Res. 9, 00694–02022. https://doi.org/10.1183/23120541.00694-2022 (2023).
doi: 10.1183/23120541.00694-2022
pubmed: 37228294
pmcid: 10204848
Raqib, R. et al. Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic. Proc. Natl. Acad. Sci. U.S.A. 103, 9178–9183. https://doi.org/10.1073/pnas.0602888103 (2006).
doi: 10.1073/pnas.0602888103
pubmed: 16740661
pmcid: 1482586
Sechet, E., Telford, E., Bonamy, C., Sansonetti, P. J. & Sperandio, B. Natural molecules induce and synergize to boost expression of the human antimicrobial peptide beta-defensin-3. Proc. Natl. Acad. Sci. U.S.A. 115, E9869–E9878. https://doi.org/10.1073/pnas.1805298115 (2018).
doi: 10.1073/pnas.1805298115
pubmed: 30275324
pmcid: 6196494
Chantret, I. et al. Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: Evidence for glucose-dependent negative regulation. J. Cell. Sci. 107(1), 213–225. https://doi.org/10.1242/jcs.107.1.213 (1994).
doi: 10.1242/jcs.107.1.213
pubmed: 8175910
Miquel, S. et al. Complete genome sequence of Crohn’s disease-associated adherent-invasive E. coli strain LF82. PLoS One 5, e12714. https://doi.org/10.1371/journal.pone.0012714 (2010).
doi: 10.1371/journal.pone.0012714
pubmed: 20862302
pmcid: 2941450
Tsakalidou, E. et al. Identification of streptococci from Greek Kasseri cheese and description of Streptococcus macedonicus sp. nov. Int. J. Syst. Bacteriol. 48(2), 519–527. https://doi.org/10.1099/00207713-48-2-519 (1998).
doi: 10.1099/00207713-48-2-519
pubmed: 9731293