Protein arginine deiminase 2 (PAD2) modulates the polarization of THP-1 macrophages to the anti-inflammatory M2 phenotype.
Citrullination
Macrophage polarization
Posttranslational modifications
Protein arginine deiminases
Proteomics
Journal
Journal of inflammation (London, England)
ISSN: 1476-9255
Titre abrégé: J Inflamm (Lond)
Pays: England
ID NLM: 101232234
Informations de publication
Date de publication:
18 Nov 2022
18 Nov 2022
Historique:
received:
30
06
2022
accepted:
31
10
2022
entrez:
19
11
2022
pubmed:
20
11
2022
medline:
20
11
2022
Statut:
epublish
Résumé
Macrophages are effector cells of the innate immune system that undergo phenotypical changes in response to organ injury and repair. These cells are most often classified as proinflammatory M1 and anti-inflammatory M2 macrophages. Protein arginine deiminase (PAD), which catalyses the irreversible conversion of protein-bound arginine into citrulline, is expressed in macrophages. However, the substrates of PAD and its role in immune cells remain unclear. This study aimed to investigate the role of PAD in THP-1 macrophage polarization to the M1 and M2 phenotypes and identify the citrullinated proteins and modified arginines that are associated with this biological switch using mass spectrometry. Our study showed that PAD2 and, to a lesser extent, PAD1 and PAD4 were predominantly expressed in M1 macrophages. We showed that inhibiting PAD expression with BB-Cl-amidine decreased macrophage polarization to the M1 phenotype (TNF-α, IL-6) and increased macrophage polarization to the M2 phenotype (MRC1, ALOX15). This process was mediated by the downregulation of proteins involved in the NF-κβ pathway. Silencing PAD2 confirmed the activation of M2 macrophages by increasing the antiviral innate immune response and interferon signalling. A total of 192 novel citrullination sites associated with inflammation, cell death and DNA/RNA processing pathways were identified in M1 and M2 macrophages. We showed that inhibiting PAD activity using a pharmacological inhibitor or silencing PAD2 with PAD2 siRNA shifted the activation of macrophages towards the M2 phenotype, which can be crucial for designing novel macrophage-mediated therapeutic strategies. We revealed a major citrullinated proteome and its rearrangement following macrophage polarization, which after further validation could lead to significant clinical benefits for the treatment of inflammation and autoimmune diseases.
Sections du résumé
BACKGROUND
BACKGROUND
Macrophages are effector cells of the innate immune system that undergo phenotypical changes in response to organ injury and repair. These cells are most often classified as proinflammatory M1 and anti-inflammatory M2 macrophages. Protein arginine deiminase (PAD), which catalyses the irreversible conversion of protein-bound arginine into citrulline, is expressed in macrophages. However, the substrates of PAD and its role in immune cells remain unclear. This study aimed to investigate the role of PAD in THP-1 macrophage polarization to the M1 and M2 phenotypes and identify the citrullinated proteins and modified arginines that are associated with this biological switch using mass spectrometry.
RESULTS
RESULTS
Our study showed that PAD2 and, to a lesser extent, PAD1 and PAD4 were predominantly expressed in M1 macrophages. We showed that inhibiting PAD expression with BB-Cl-amidine decreased macrophage polarization to the M1 phenotype (TNF-α, IL-6) and increased macrophage polarization to the M2 phenotype (MRC1, ALOX15). This process was mediated by the downregulation of proteins involved in the NF-κβ pathway. Silencing PAD2 confirmed the activation of M2 macrophages by increasing the antiviral innate immune response and interferon signalling. A total of 192 novel citrullination sites associated with inflammation, cell death and DNA/RNA processing pathways were identified in M1 and M2 macrophages.
CONCLUSIONS
CONCLUSIONS
We showed that inhibiting PAD activity using a pharmacological inhibitor or silencing PAD2 with PAD2 siRNA shifted the activation of macrophages towards the M2 phenotype, which can be crucial for designing novel macrophage-mediated therapeutic strategies. We revealed a major citrullinated proteome and its rearrangement following macrophage polarization, which after further validation could lead to significant clinical benefits for the treatment of inflammation and autoimmune diseases.
Identifiants
pubmed: 36401289
doi: 10.1186/s12950-022-00317-8
pii: 10.1186/s12950-022-00317-8
pmc: PMC9675280
doi:
Types de publication
Journal Article
Langues
eng
Pagination
20Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL111362
Pays : United States
Informations de copyright
© 2022. The Author(s).
Références
Front Immunol. 2018 Sep 18;9:2036
pubmed: 30279690
Circ Res. 2010 May 28;106(10):1559-69
pubmed: 20508200
FASEB J. 2021 Sep;35(9):e21866
pubmed: 34416031
Cell. 2005 Sep 9;122(5):669-82
pubmed: 16125763
Ann Rheum Dis. 2015 Dec;74(12):2199-206
pubmed: 25104775
Front Immunol. 2013 Mar 18;4:67
pubmed: 23508122
Mol Cell. 2017 Aug 3;67(3):387-399.e5
pubmed: 28712728
J Biol Chem. 2013 Sep 27;288(39):28324-30
pubmed: 23943623
Int J Mol Sci. 2020 Aug 10;21(16):
pubmed: 32785008
J Biol Chem. 2001 Jun 22;276(25):22215-22
pubmed: 11297557
Methods Mol Biol. 2022;2420:107-126
pubmed: 34905169
J Biochem. 2009 Oct;146(4):471-9
pubmed: 19564157
J Proteome Res. 2009 Dec;8(12):5674-8
pubmed: 19848406
J Proteomics. 2014 Sep 23;109:322-31
pubmed: 25063446
Nat Biotechnol. 2014 Mar;32(3):223-6
pubmed: 24727771
Mol Cell Biol. 2000 Jul;20(13):4532-42
pubmed: 10848580
Lupus. 2011 Mar;20(3):274-89
pubmed: 21362751
Cytokine. 2016 Jul;83:171-175
pubmed: 27152709
J Immunol Res. 2019 Nov 25;2019:7592851
pubmed: 31886309
QJM. 2018 Nov 01;111(11):769-778
pubmed: 30016493
J Am Soc Mass Spectrom. 2020 Jul 1;31(7):1410-1421
pubmed: 32463229
Nat Cell Biol. 2006 Apr;8(4):398-406
pubmed: 16547522
J Biol Chem. 2010 Dec 17;285(51):39655-62
pubmed: 20937835
J Proteome Res. 2019 May 3;18(5):2270-2278
pubmed: 30990720
Nat Rev Immunol. 2008 Dec;8(12):958-69
pubmed: 19029990
Sci Transl Med. 2021 Mar 17;13(585):
pubmed: 33731433
Nat Rev Immunol. 2003 Jan;3(1):23-35
pubmed: 12511873
Mol Cell Proteomics. 2015 May;14(5):1400-10
pubmed: 25724911
Biochem Pharmacol. 2019 Jul;165:152-169
pubmed: 30910693
Int Immunopharmacol. 2014 Nov;23(1):37-45
pubmed: 25130606
J Immunol Res. 2019 Dec 1;2019:4354786
pubmed: 31886303
PLoS Pathog. 2021 May 14;17(5):e1009597
pubmed: 33989349
Oncogene. 2016 Dec 1;35(48):6246-6261
pubmed: 27157615
J Rheumatol Suppl. 1987 Jun;14 Suppl 13:78-82
pubmed: 2956421
Nature. 2020 Jan;577(7788):109-114
pubmed: 31827280
Int J Mol Sci. 2018 Mar 13;19(3):
pubmed: 29533975
Nature. 2009 Oct 8;461(7265):788-92
pubmed: 19776740
J Exp Med. 2002 Jun 17;195(12):1647-52
pubmed: 12070292
Immunity. 2017 Mar 21;46(3):393-404
pubmed: 28314590
J Am Heart Assoc. 2014 Nov 12;3(6):e001329
pubmed: 25392542
Kidney Blood Press Res. 2019;44(1):62-71
pubmed: 30808838
J Immunol. 2019 Aug 15;203(4):795-800
pubmed: 31292215
J Immunol Res. 2018 Jan 14;2018:8917804
pubmed: 29507865
Nucleic Acids Res. 2014 Feb;42(4):2708-24
pubmed: 24311566
FASEB J. 2014 Aug;28(8):3758-68
pubmed: 24823363
Cell. 2009 Jun 12;137(6):1112-23
pubmed: 19524513
Pflugers Arch. 2022 Aug;474(8):949-962
pubmed: 35403906
Clin Exp Rheumatol. 2019 Nov-Dec;37(6):929-936
pubmed: 30789148
Cell Signal. 2021 Oct;86:110075
pubmed: 34229086
Mol Cell Proteomics. 2020 Feb;19(2):421-430
pubmed: 31888964
Nat Commun. 2017 Oct 16;8(1):950
pubmed: 29038465
Curr Drug Targets. 2015;16(7):700-10
pubmed: 25642720
Immunol Lett. 2021 Nov;239:23-31
pubmed: 34418490
Nat Methods. 2016 Sep;13(9):731-40
pubmed: 27348712
Aging (Albany NY). 2020 Oct 27;12(20):20432-20444
pubmed: 33109771
Biochemistry. 2006 Oct 3;45(39):11727-36
pubmed: 17002273
Prog Chem Org Nat Prod. 2017;106:203-240
pubmed: 28762090
Cells. 2021 Aug 21;10(8):
pubmed: 34440923
Signal Transduct Target Ther. 2017;2:
pubmed: 29158945
J Clin Invest. 2001 Jan;107(1):7-11
pubmed: 11134171
Cell. 2000 Oct 13;103(2):351-61
pubmed: 11057907
Int J Biochem Cell Biol. 2006;38(10):1662-77
pubmed: 16730216
J Immunol Methods. 2020 Mar;478:112721
pubmed: 32033786