Protein arginine methyltransferase 5 mediates THP‑1‑derived macrophage activation dependent on NF‑κB in endometriosis

Chai,Xiaoshan; Wu,Xianqing; He,Ling; Ding,Hui

doi:10.3892/etm.2021.10436

Protein arginine methyltransferase 5 mediates THP‑1‑derived macrophage activation dependent on NF‑κB in endometriosis

Authors:
- Xiaoshan Chai
- Xianqing Wu
- Ling He
- Hui Ding
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Affiliations: Department of Obstetrics and Gynecology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: July 15, 2021 https://doi.org/10.3892/etm.2021.10436
Article Number: 1003
Copyright: © Chai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Macrophage‑induced inflammation is a major factor in the pathogenesis of endometriosis. The underlying mechanisms, however, remain largely unknown. TNF‑α, IL‑6, IL‑10 and C‑C motif chemokine 20 (CCL20) levels in endometrial extracts were determined using Luminex cytokine kits. Additionally, protein arginine methyltransferase 5 (PRMT5) levels were measured using reverse transcription‑quantitative PCR and western blotting. IL‑6 and IP‑10 levels in cells were measured using ELISA kits. In the present study, it was revealed that PRMT5 expression at both the mRNA and protein levels in THP‑1‑derived macrophages was significantly decreased following treatment with serum or extracts of endometrium from patients with endometriosis in the presence of lipopolysaccharide, compared with that in control cells, suggesting a possible role for macrophage‑derived PRMT5 in mediating the interaction between macrophages and endometrium in endometriosis. Mechanistically, macrophage PRMT5 expression was regulated in an NF‑κB‑dependent and Smad2/3‑independent manner, indicating that PRMT5 is a downstream target of NF‑κB. Importantly, macrophage‑derived PRMT5 was required for macrophage activation in endometriosis, as evidenced by the PRMT5‑dependent secretion of IL‑6 and IFN‑γ‑induced protein 10 from THP‑1‑derived macrophages. The present study identified NF‑κB‑dependent PRMT5 as a novel regulator of macrophage activation in endometriosis. Targeting PRMT5 in macrophages may be a potential therapeutic strategy against endometriosis.

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1	Mehedintu C, Plotogea MN, Ionescu S and Antonovici M: Endometriosis still a challenge. J Med Life. 7:349–357. 2014.PubMed/NCBI
2	Scheerer C, Bauer P, Chiantera V, Sehouli J, Kaufmann A and Mechsner S: Characterization of endometriosis-associated immune cell infiltrates (EMaICI). Arch Gynecol Obstet. 294:657–664. 2016.PubMed/NCBI View Article : Google Scholar
3	Bacci M, Capobianco A, Monno A, Cottone L, Di Puppo F, Camisa B, Mariani M, Brignole C, Ponzoni M, Ferrari S, et al: Macrophages are alternatively activated in patients with endometriosis and required for growth and vascularization of lesions in a mouse model of disease. Am J Pathol. 175:547–556. 2009.PubMed/NCBI View Article : Google Scholar
4	Ahn SH, Edwards AK, Singh SS, Young SL, Lessey BA and Tayade C: IL-17A contributes to the pathogenesis of endometriosis by triggering proinflammatory cytokines and angiogenic growth factors. J Immunol. 195:2591–2600. 2015.PubMed/NCBI View Article : Google Scholar
5	Antsiferova YS, Sotnikova NY, Posiseeva LV and Shor AL: Changes in the T-helper cytokine profile and in lymphocyte activation at the systemic and local levels in women with endometriosis. Fertil Steril. 84:1705–1711. 2005.PubMed/NCBI View Article : Google Scholar
6	Gogacz M, Winkler I, Bojarska-Junak A, Tabarkiewicz J, Semczuk A, Rechberger T and Adamiak A: Increased percentage of Th17 cells in peritoneal fluid is associated with severity of endometriosis. J Reprod Immunol. 117:39–44. 2016.PubMed/NCBI View Article : Google Scholar
7	Izumi G, Koga K, Takamura M, Makabe T, Satake E, Takeuchi A, Taguchi A, Urata Y, Fujii T and Osuga Y: Involvement of immune cells in the pathogenesis of endometriosis. J Obstet Gynaecol Res. 44:191–198. 2018.PubMed/NCBI View Article : Google Scholar
8	Berbic M, Schulke L, Markham R, Tokushige N, Russell P and Fraser IS: Macrophage expression in endometrium of women with and without endometriosis. Hum Reprod. 24:325–332. 2009.PubMed/NCBI View Article : Google Scholar
9	Kim TH, Yoo JY, Choi KC, Shin JH, Leach RE, Fazleabas AT, Young SL, Lessey BA, Yoon HG and Jeong JW and Jeong JW: Loss of HDAC3 results in nonreceptive endometrium and female infertility. Sci Transl Med. 11(eaaf7533)2019.PubMed/NCBI View Article : Google Scholar
10	Bozdag G: Recurrence of endometriosis: Risk factors, mechanisms and biomarkers. Womens Health (Lond). 11:693–699. 2015.PubMed/NCBI View Article : Google Scholar
11	Gheorghisan-Galateanu AA and Gheorghiu ML: Hormonal therapy in women of reproductive age with endometriosis: An update. Acta Endocrinol (Buchar). 15:276–281. 2019.PubMed/NCBI View Article : Google Scholar
12	Karkhanis V, Hu YJ, Baiocchi RA, Imbalzano AN and Sif S: Versatility of PRMT5-induced methylation in growth control and development. Trends Biochem Sci. 36:633–641. 2011.PubMed/NCBI View Article : Google Scholar
13	Bandyopadhyay S, Harris DP, Adams GN, Lause GE, McHugh A, Tillmaand EG, Money A, Willard B, Fox PL and Dicorleto PE: HOXA9 methylation by PRMT5 is essential for endothelial cell expression of leukocyte adhesion molecules. Mol Cell Biol. 32:1202–1213. 2012.PubMed/NCBI View Article : Google Scholar
14	Webb LM, Amici SA, Jablonski KA, Savardekar H, Panfil AR, Li L, Zhou W, Peine K, Karkhanis V, Bachelder EM, et al: PRMT5-selective inhibitors suppress inflammatory T cell responses and experimental autoimmune encephalomyelitis. J Immunol. 198:1439–1451. 2017.PubMed/NCBI View Article : Google Scholar
15	Zheng Y, Huang L, Ge W, Yang M, Ma Y, Xie G, Wang W, Bian B, Li L, Nie H and Shen L: Protein arginine methyltransferase 5 inhibition upregulates Foxp3(+) regulatory T cells frequency and function during the ulcerative colitis. Front Immunol. 8(596)2017.PubMed/NCBI View Article : Google Scholar
16	Bian X, Li B, Yang J, Ma K, Sun M, Zhang C and Fu X: Regenerative and protective effects of dMSC-sEVs on high-glucose-induced senescent fibroblasts by suppressing RAGE pathway and activating Smad pathway. Stem Cell Res Ther. 11(166)2020.PubMed/NCBI View Article : Google Scholar
17	Johnson NP, Hummelshoj L, Adamson GD, Keckstein J, Taylor HS, Abrao MS, Bush D, Kiesel L, Tamimi R, Sharpe-Timms KL, et al: World endometriosis society consensus on the classification of endometriosis. Hum Reprod. 32:315–324. 2017.PubMed/NCBI View Article : Google Scholar
18	Fischer AH, Jacobson KA, Rose J and Zeller R: Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc. 2008(pdb.prot4986)2008.PubMed/NCBI View Article : Google Scholar
19	Xie Q, He H, Wu YH, Zou LJ, She XL, Xia XM and Wu XQ: Eutopic endometrium from patients with endometriosis modulates the expression of CD36 and SIRP-α in peritoneal macrophages. J Obstet Gynaecol Res. 45:1045–1057. 2019.PubMed/NCBI View Article : Google Scholar
20	Wu XQ, Fang XL, Lin QH, Huang FY and Xia XM: VEGF production and its receptor expression of MPhis in peritoneal fluid. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 19:462–465. 2003.PubMed/NCBI(In Chinese).
21	McLaren J, Prentice A, Charnock-Jones DS, Millican SA, Müller KH, Sharkey AM and Smith SK: Vascular endothelial growth factor is produced by peritoneal fluid macrophages in endometriosis and is regulated by ovarian steroids. J Clin Invest. 98:482–489. 1996.PubMed/NCBI View Article : Google Scholar
22	Dasgupta S, Bhattacharya-Chatterjee M, O'Malley BW Jr and Chatterjee SK: Inhibition of NK cell activity through TGF-beta 1 by down-regulation of NKG2D in a murine model of head and neck cancer. J Immunol. 175:5541–5550. 2005.PubMed/NCBI View Article : Google Scholar
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
24	Chen D, Zeng S, Huang M, Xu H, Liang L and Yang X: Role of protein arginine methyltransferase 5 in inflammation and migration of fibroblast-like synoviocytes in rheumatoid arthritis. J Cell Mol Med. 21:781–790. 2017.PubMed/NCBI View Article : Google Scholar
25	Li S, Fu X, Wu T, Yang L, Hu C and Wu R: Role of interleukin-6 and its receptor in endometriosis. Med Sci Monit. 23:3801–3807. 2017.PubMed/NCBI View Article : Google Scholar
26	Ulukus M and Arici A: Immunology of endometriosis. Minerva Ginecol. 57:237–248. 2005.PubMed/NCBI
27	Haber E, Danenberg HD, Koroukhov N, Ron-El R, Golomb G and Schachter M: Peritoneal macrophage depletion by liposomal bisphosphonate attenuates endometriosis in the rat model. Hum Reprod. 24:398–407. 2009.PubMed/NCBI View Article : Google Scholar
28	Liu G and Yang H: Modulation of macrophage activation and programming in immunity. J Cell Physiol. 228:502–512. 2013.PubMed/NCBI View Article : Google Scholar
29	Li MZ, Wu YH, Ali M, Wu XQ and Nie MF: Endometrial stromal cells treated by tumor necrosis factor-α stimulate macrophages polarized toward M2 via interleukin-6 and monocyte chemoattractant protein-1. J Obstet Gynaecol Res. 46:293–301. 2020.PubMed/NCBI View Article : Google Scholar
30	Doucet M, Jayaraman S, Swenson E, Tusing B, Weber KL and Kominsky SL: CCL20/CCR6 signaling regulates bone mass accrual in mice. J Bone Miner Res. 31:1381–1390. 2016.PubMed/NCBI View Article : Google Scholar
31	Fan Z, Kong X, Xia J, Wu X, Li H, Xu H, Fang M and Xu Y: The arginine methyltransferase PRMT5 regulates CIITA-dependent MHC II transcription. Biochim Biophys Acta. 1859:687–696. 2016.PubMed/NCBI View Article : Google Scholar
32	Ding Q, Chen J, Wei X, Sun W, Mai J, Yang Y and Xu Y: RAFTsomes containing epitope-MHC-II complexes mediated CD4⁺ T cell activation and antigen-specific immune responses. Pharm Res. 30:60–69. 2013.PubMed/NCBI View Article : Google Scholar
33	González-Ramos R, Defrère S and Devoto L: Nuclear factor-kappaB: A main regulator of inflammation and cell survival in endometriosis pathophysiology. Fertil Steril. 98:520–528. 2012.PubMed/NCBI View Article : Google Scholar
34	Wei H, Wang B, Miyagi M, She Y, Gopalan B, Huang DB, Ghosh G, Stark GR and Lu T: PRMT5 dimethylates R30 of the p65 subunit to activate NF-κB. Proc Natl Acad Sci USA. 110:13516–13521. 2013.PubMed/NCBI View Article : Google Scholar
35	Chan-Penebre E, Kuplast KG, Majer CR, Boriack-Sjodin PA, Wigle TJ, Johnston LD, Rioux N, Munchhof MJ, Jin L, Jacques SL, et al: A selective inhibitor of PRMT5 with in vivo and in vitro potency in MCL models. Nat Chem Biol. 11:432–437. 2015.PubMed/NCBI View Article : Google Scholar
36	Harris DP, Bandyopadhyay S, Maxwell TJ, Willard B and DiCorleto PE: Tumor necrosis factor (TNF)-α induction of CXCL10 in endothelial cells requires protein arginine methyltransferase 5 (PRMT5)-mediated nuclear factor (NF)-κB p65 methylation. J Biol Chem. 289:15328–15339. 2014.PubMed/NCBI View Article : Google Scholar
37	Poli-Neto OB, Meola J, Rosa-E-Silva JC and Tiezzi D: Transcriptome meta-analysis reveals differences of immune profile between eutopic endometrium from stage I-II and III-IV endometriosis independently of hormonal milieu. Sci Rep. 10(313)2020.PubMed/NCBI View Article : Google Scholar
38	Nagarsheth N, Wicha MS and Zou W: Chemokines in the cancer microenvironment and their relevance in cancer immunotherapy. Nat Rev Immunol. 17:559–572. 2017.PubMed/NCBI View Article : Google Scholar