Impaired function of epithelial plakophilin-2 is associated with periodontal disease.
PKP2
desmosome
epithelium
gene expression
periodontal disease
Journal
Journal of periodontal research
ISSN: 1600-0765
Titre abrégé: J Periodontal Res
Pays: United States
ID NLM: 0055107
Informations de publication
Date de publication:
Dec 2021
Dec 2021
Historique:
received:
30
06
2021
accepted:
13
07
2021
pubmed:
10
8
2021
medline:
20
11
2021
entrez:
9
8
2021
Statut:
ppublish
Résumé
Plakophilin-2 (PKP2) is an intracellular desmosomal anchoring protein that has been implicated in a genome-wide association study, in which genetic variants of PKP2 are associated with Porphyromonas gingivalis (P.gingivalis) -dominant periodontal dysbiosis. In this study, we compared the ex vivo PKP2 expression in periodontitis gingival biopsies to periodontitis-free subjects and assessed the in vitro role of PKP2 in gingival epithelial barrier function and the mechanism by which P.gingivalis modulates PKP2 expression. Using reverse transcription quantitative real-time PCR (RT-qPCR), we determined PKP2 mRNA expression levels in gingival biopsies collected from 11 periodontally healthy, 10 experimental gingivitis, and 10 chronic periodontitis subjects. PKP2 protein expression in gingival biopsies was detected by immunohistochemistry. We then challenged primary gingival epithelial cells with bacteria including P.gingivalis, Campylobacter rectus, and various Toll-like receptor agonists. Western blot and immunofluorescence staining were used to detect protein expression. Inhibitors blocking proteases pathways were tested for P.gingivalis-mediated PKP2 protein degradations. We also knocked down endogenous epithelial PKP2 using lentiviral short-hairpin RNA (shRNA) and evaluated cell proliferation, spreading, and barrier function. Periodontitis gingival biopsies had approximately twofold less PKP2 mRNA than did healthy controls (p < .05). PKP2 protein was predominantly expressed in gingival epithelium. In primary gingival epithelial cells, P.gingivalis challenge increased PKP2 mRNA levels, while protein expression decreased, which suggests that P.gingivalis has a protein degradation mechanism. Cysteine proteases inhibitors greatly attenuated P.gingivalis-mediated PKP2 protein degradation. Epithelial cells with deficient PKP2 exhibited inhibited cell proliferation and spreading and failed to form monolayers. Finally, P.gingivalis impaired gingival epithelial barrier function. PKP2 appears to be critical in maintaining gingival epithelial barrier function and is susceptible to degradation by cysteine proteases produced by P.gingivalis. Our findings have identified a mechanism by which P.gingivalis impairs epithelial barrier function by promoting PKP2 degradation.
Sections du résumé
BACKGROUND AND OBJECTIVES
OBJECTIVE
Plakophilin-2 (PKP2) is an intracellular desmosomal anchoring protein that has been implicated in a genome-wide association study, in which genetic variants of PKP2 are associated with Porphyromonas gingivalis (P.gingivalis) -dominant periodontal dysbiosis. In this study, we compared the ex vivo PKP2 expression in periodontitis gingival biopsies to periodontitis-free subjects and assessed the in vitro role of PKP2 in gingival epithelial barrier function and the mechanism by which P.gingivalis modulates PKP2 expression.
MATERIAL AND METHODS
METHODS
Using reverse transcription quantitative real-time PCR (RT-qPCR), we determined PKP2 mRNA expression levels in gingival biopsies collected from 11 periodontally healthy, 10 experimental gingivitis, and 10 chronic periodontitis subjects. PKP2 protein expression in gingival biopsies was detected by immunohistochemistry. We then challenged primary gingival epithelial cells with bacteria including P.gingivalis, Campylobacter rectus, and various Toll-like receptor agonists. Western blot and immunofluorescence staining were used to detect protein expression. Inhibitors blocking proteases pathways were tested for P.gingivalis-mediated PKP2 protein degradations. We also knocked down endogenous epithelial PKP2 using lentiviral short-hairpin RNA (shRNA) and evaluated cell proliferation, spreading, and barrier function.
RESULTS
RESULTS
Periodontitis gingival biopsies had approximately twofold less PKP2 mRNA than did healthy controls (p < .05). PKP2 protein was predominantly expressed in gingival epithelium. In primary gingival epithelial cells, P.gingivalis challenge increased PKP2 mRNA levels, while protein expression decreased, which suggests that P.gingivalis has a protein degradation mechanism. Cysteine proteases inhibitors greatly attenuated P.gingivalis-mediated PKP2 protein degradation. Epithelial cells with deficient PKP2 exhibited inhibited cell proliferation and spreading and failed to form monolayers. Finally, P.gingivalis impaired gingival epithelial barrier function.
CONCLUSIONS
CONCLUSIONS
PKP2 appears to be critical in maintaining gingival epithelial barrier function and is susceptible to degradation by cysteine proteases produced by P.gingivalis. Our findings have identified a mechanism by which P.gingivalis impairs epithelial barrier function by promoting PKP2 degradation.
Identifiants
pubmed: 34368962
doi: 10.1111/jre.12918
pmc: PMC8627832
mid: NIHMS1725091
doi:
Substances chimiques
PKP2 protein, human
0
Plakophilins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1046-1057Subventions
Organisme : NIDCR NIH HHS
ID : K99 DE027086
Pays : United States
Organisme : NIDCR NIH HHS
ID : R00 DE027086
Pays : United States
Organisme : NIDCR NIH HHS
ID : R90DE022527
Pays : United States
Organisme : NIDCR NIH HHS
ID : R00DE027086
Pays : United States
Informations de copyright
© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Références
Iran J Pathol. 2015 Summer;10(3):214-20
pubmed: 26351487
Front Cell Infect Microbiol. 2018 Jan 24;8:10
pubmed: 29416987
Hum Mol Genet. 2016 May 15;25(10):2113-2129
pubmed: 26962152
J Periodontol. 2016 Feb;87(2):193-202
pubmed: 26430925
Am J Pathol. 2018 Feb;188(2):404-416
pubmed: 29154960
Infect Immun. 1998 Nov;66(11):5337-43
pubmed: 9784541
Curr Opin Cell Biol. 2004 Oct;16(5):536-43
pubmed: 15363804
J Transl Med. 2010 Nov 04;8:110
pubmed: 21047437
Indian J Dent Res. 2010 Jul-Sep;21(3):396-401
pubmed: 20930352
Biochim Biophys Acta. 2008 Mar;1778(3):572-87
pubmed: 17854763
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
J Periodontol. 2017 Jul;88(7):663-672
pubmed: 28387608
J Biol Chem. 2004 Feb 13;279(7):5268-77
pubmed: 14630910
J Periodontol. 2014 Aug;85(8):e313-22
pubmed: 24527855
Periodontol 2000. 2018 Oct;78(1):174-184
pubmed: 30198133
J Pathol. 2000 Sep;192(1):58-66
pubmed: 10951401
J Periodontol. 1965 May-Jun;36:177-87
pubmed: 14296927
J Immunol. 2003 Dec 1;171(11):6164-72
pubmed: 14634132
J Invest Dermatol. 2014 Jan;134(1):112-122
pubmed: 23884246
Circulation. 2006 Apr 4;113(13):1650-8
pubmed: 16567567
J Periodontol. 2007 Nov;78(11):2197-202
pubmed: 17970688
Infect Immun. 2002 Jan;70(1):257-67
pubmed: 11748191
Nat Med. 2000 Nov;6(11):1275-7
pubmed: 11062541
Crit Rev Oral Biol Med. 1991;2(1):13-32
pubmed: 1912142
J Periodontal Res. 1988 Mar;23(2):81-6
pubmed: 2967369
Mol Cell Biol. 2014 Oct;34(20):3843-54
pubmed: 25113560
Infect Immun. 1998 Sep;66(9):4108-14
pubmed: 9712755
J Dent Res. 2019 Sep;98(10):1150-1158
pubmed: 31340691
J Cell Biol. 2004 Oct 11;167(1):149-60
pubmed: 15479741
J Periodontal Res. 2006 Aug;41(4):322-8
pubmed: 16827727
Pesqui Odontol Bras. 2003 Jan-Mar;17(1):41-5
pubmed: 12908058
Periodontol 2000. 2020 Feb;82(1):129-142
pubmed: 31850627
Hum Mol Genet. 2013 Jun 1;22(11):2312-24
pubmed: 23459936
Nat Commun. 2017 Jul 24;8(1):106
pubmed: 28740174
FEMS Microbiol Lett. 2003 Apr 25;221(2):181-5
pubmed: 12725924
PLoS One. 2013 Nov 12;8(11):e78541
pubmed: 24265696
Tissue Barriers. 2017 Oct 2;5(4):e1343172
pubmed: 28686506
J Am Soc Nephrol. 2006 Jul;17(7):1807-19
pubmed: 16738015
J Invest Dermatol. 2003 Aug;121(2):383-9
pubmed: 12880431
BMC Microbiol. 2009 May 27;9:107
pubmed: 19473524
JDR Clin Trans Res. 2016 Jul;1(2):163-170
pubmed: 28459102
Curr Drug Targets. 2011 Mar 1;12(3):322-31
pubmed: 20955149
Ann Periodontol. 1999 Dec;4(1):1-6
pubmed: 10863370
J Periodontal Res. 2014 Apr;49(2):253-9
pubmed: 23713517
Nat Commun. 2018 Sep 11;9(1):3686
pubmed: 30206230
J Dent Res. 2010 Feb;89(2):133-7
pubmed: 20042743
J Periodontal Res. 2011 Jun;46(3):345-53
pubmed: 21382035
Nat Rev Mol Cell Biol. 2000 Dec;1(3):208-16
pubmed: 11252896
Nat Rev Dis Primers. 2017 May 11;3:17026
pubmed: 28492232
J Periodontol. 2009 Dec;80(12):1963-82
pubmed: 19961380