N6‑methyladenosine methyltransferase METTL14 is associated with macrophage polarization in rheumatoid arthritis

Zhu,Ziheng; Wan,Lei

doi:10.3892/etm.2024.12664

N6‑methyladenosine methyltransferase METTL14 is associated with macrophage polarization in rheumatoid arthritis

Authors:
- Ziheng Zhu
- Lei Wan
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Affiliations: Department of Rheumatology and Immunology, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, Anhui 230031, P.R. China
Published online on: July 25, 2024 https://doi.org/10.3892/etm.2024.12664
Article Number: 375
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Rheumatoid arthritis (RA) is largely caused by the inflammatory response triggered by macrophage polarization. Through epigenetic reprogramming, the inflammatory state of macrophages can be modified. Macrophage polarization is associated with the RNA epigenetic alteration N6‑methyladenosine (m6A) RNA methylation. However, the specific function and underlying mechanisms of m6A methylation in the role of macrophage polarization in RA remain to be elucidated. The mRNA expression levels of m6A methylase genes and signaling pathway components associated with RA macrophages were determined in the present study using reverse‑transcription quantitative PCR. Methyltransferase 14 (METTL14) protein expression levels were determined using western blot analysis, and the levels of specific cellular secretion factors were determined using ELISA and flow cytometry. The results of the present study demonstrated that elevated METTL14 expression was associated with joint tenderness, and METTL14 expression was positively correlated with both C‑reactive protein and rheumatoid factor expression levels. Moreover, METTL14 exhibited potential in the prediction of visual analogue scale. Pro‑inflammatory cytokines (TNF‑α) and M1 macrophage markers (CD68⁺CD86⁺) were also positively associated with METTL14 expression. The results of the Kyoto Encyclopedia of Genes and Genomes analysis revealed that METTL14 was strongly associated with the MAPK signaling pathway. Notably, JNK and ERK2 exhibited a positive correlation with the M1 macrophage marker, CD68⁺CD86⁺, which was positively associated with the pro‑inflammatory factor, TNF‑α. JNK and ERK2 expression levels were markedly increased in the METTL14 high‑expression group, compared with in the low‑expression group; however, p38 and ERK1 expression levels were not significantly different between these groups. Collectively, the results of the present study demonstrated that METTL14 expression was significantly increased in the peripheral blood and synovial tissue of patients with RA, highlighting the potential association with both immunoinflammatory markers and clinical symptoms. In addition, it was suggested that METTL14 may exacerbate the downstream inflammatory response, through mediating macrophage polarization via the MAPK pathway.

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1	Wan L, Liu J, Huang C, Zhu Z, Wang K, Sun G, Zhu L and Hu Z: Comprehensive analysis and functional characteristics of differential expression of N6-methyladenosine methylation modification in the whole transcriptome of rheumatoid arthritis. Mediators Inflamm. 2022(4766992)2022.PubMed/NCBI View Article : Google Scholar
2	Liu S, Ma H, Zhang H, Deng C and Xin P: Recent advances on signaling pathways and their inhibitors in rheumatoid arthritis. Clin Immunol. 230(108793)2021.PubMed/NCBI View Article : Google Scholar
3	Alivernini S, Firestein GS and McInnes IB: The pathogenesis of rheumatoid arthritis. Immunity. 55:2255–2270. 2022.PubMed/NCBI View Article : Google Scholar
4	Kurowska-Stolarska M and Alivernini S: Synovial tissue macrophages in joint homeostasis, rheumatoid arthritis and disease remission. Nat Rev Rheumatol. 18:384–397. 2022.PubMed/NCBI View Article : Google Scholar
5	Jang S, Kwon EJ and Lee JJ: Rheumatoid arthritis: Pathogenic roles of diverse immune cells. Int J Mol Sci. 23(905)2022.PubMed/NCBI View Article : Google Scholar
6	Jiang P and Li X: Regulatory mechanism of lncRNAs in M1/M2 macrophages polarization in the diseases of different etiology. Front Immunol. 13(835932)2022.PubMed/NCBI View Article : Google Scholar
7	Cutolo M, Campitiello R, Gotelli E and Soldano S: The role of M1/M2 macrophage polarization in rheumatoid arthritis synovitis. Front Immunol. 13(867260)2022.PubMed/NCBI View Article : Google Scholar
8	Hasegawa T and Ishii M: Pathological osteoclasts and precursor macrophages in inflammatory arthritis. Front Immunol. 13(867368)2022.PubMed/NCBI View Article : Google Scholar
9	Gao L and Lu Q: The critical importance of epigenetics in autoimmune-related skin diseases. Front Med. 17:43–57. 2023.PubMed/NCBI View Article : Google Scholar
10	Nair N, Barton A and Wilson AG: Cell-specific epigenetic drivers of pathogenesis in rheumatoid arthritis. Epigenomics. 13:549–560. 2021.PubMed/NCBI View Article : Google Scholar
11	Chen Q, Li H, Liu Y and Zhao M: Epigenetic regulation of immune and inflammatory responses in rheumatoid arthritis. Front Immunol. 13(881191)2022.PubMed/NCBI View Article : Google Scholar
12	Jain N, Lord JM and Vogel V: Mechanoimmunology: Are inflammatory epigenetic states of macrophages tuned by biophysical factors? APL Bioeng. 6(031502)2022.PubMed/NCBI View Article : Google Scholar
13	Ghiboub M, Koster J, Craggs PD, Li Yim AYF, Shillings A, Hutchinson S, Bingham RP, Gatfield K, Hageman IL, Yao G, et al: Modulation of macrophage inflammatory function through selective inhibition of the epigenetic reader protein SP140. BMC Biol. 20(182)2022.PubMed/NCBI View Article : Google Scholar
14	Wu S, Li XF, Wu YY, Yin SQ, Huang C and Li J: N6-methyladenosine and rheumatoid arthritis: A comprehensive review. Front Immunol. 12(731842)2021.PubMed/NCBI View Article : Google Scholar
15	Gan L, Zhao Y, Fu Y and Chen Q: The potential role of m6A modifications on immune cells and immunotherapy. Biomed Pharmacother. 160(114343)2023.PubMed/NCBI View Article : Google Scholar
16	Elsabbagh RA, Rady M, Watzl C, Abou-Aisha K and Gad MZ: Impact of N6-methyladenosine (m⁶A) modification on immunity. Cell Commun Signal. 20(140)2022.PubMed/NCBI View Article : Google Scholar
17	Li C, Zhu M, Wang J, Wu H, Liu Y and Huang D: Role of m6A modification in immune microenvironment of digestive system tumors. Biomed Pharmacother. 164(114953)2023.PubMed/NCBI View Article : Google Scholar
18	Geng Q, Cao X, Fan D, Wang Q, Wang X, Zhang M, Zhao L, Jiao Y, Deng T, Liu H, et al: Potential medicinal value of N6-methyladenosine in autoimmune diseases and tumours. Br J Pharmacol: Jun 9, 2023 (Epub ahead of print).
19	Wang Y, Li L, Li J, Zhao B, Huang G, Li X, Xie Z and Zhou Z: The emerging role of m6A modification in regulating the immune system and autoimmune diseases. Front Cell Dev Biol. 9(755691)2021.PubMed/NCBI View Article : Google Scholar
20	Ma Z, Sugimura R and Lui KO: The role of m6A mRNA modification in normal and malignant hematopoiesis. J Leukoc Biol. 115:100–115. 2024.PubMed/NCBI View Article : Google Scholar
21	Kang K, Park SH, Chen J, Qiao Y, Giannopoulou E, Berg K, Hanidu A, Li J, Nabozny G, Kang K, et al: Interferon-γ represses M2 gene expression in human macrophages by disassembling enhancers bound by the transcription factor MAF. Immunity. 47:235–250. 2017.PubMed/NCBI View Article : Google Scholar
22	Kuriya B, Schieir O, Lin D, Xiong J, Pope J, Boire G, Haraoui B, Thorne JC, Tin D, Hitchon C, et al: Thresholds for the 28-joint disease activity score (DAS28) using C-reactive protein are lower compared to DAS28 using erythrocyte sedimentation rate in early rheumatoid arthritis. Clin Exp Rheumatol. 35:799–803. 2017.PubMed/NCBI
23	Castrejón I, Chua JR and Pincus T: A RheuMetric physician checklist to quantitate levels of inflammation, damage and distress on 0-10 visual analogue scales. Clin Exp Rheumatol. 35 (Suppl 107):S21–S25. 2017.PubMed/NCBI
24	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
25	Geng Q, Cao X, Fan D, Gu X, Zhang Q, Zhang M, Wang Z, Deng T and Xiao C: Diagnostic gene signatures and aberrant pathway activation based on m6A methylation regulators in rheumatoid arthritis. Front Immunol. 13(1041284)2022.PubMed/NCBI View Article : Google Scholar
26	Zhang X, Li X, Jia H, An G and Ni J: The m⁶A methyltransferase METTL3 modifies PGC-1α mRNA promoting mitochondrial dysfunction and oxLDL-induced inflammation in monocytes. J Biol Chem. 297(101058)2021.PubMed/NCBI View Article : Google Scholar
27	Saeki N and Imai Y: Crosstalk between synovial macrophages and fibroblasts in rheumatoid arthritis. Histol Histopathol. 38:1231–1238. 2023.PubMed/NCBI View Article : Google Scholar
28	Boutet MA, Courties G, Nerviani A, Le Goff B, Apparailly F, Pitzalis C and Blanchard F: Novel insights into macrophage diversity in rheumatoid arthritis synovium. Autoimmun Rev. 20(102758)2021.PubMed/NCBI View Article : Google Scholar
29	Tong J, Flavell RA and Li HB: RNA m⁶A modification and its function in diseases. Front Med. 12:481–489. 2018.PubMed/NCBI View Article : Google Scholar
30	Dou X, Huang L, Xiao Y, Liu C, Li Y, Zhang X, Yu L, Zhao R, Yang L, Chen C, et al: METTL14 is a chromatin regulator independent of its RNA N6-methyladenosine methyltransferase activity. Protein Cell. 14:683–697. 2023.PubMed/NCBI View Article : Google Scholar
31	Li X, Xu X, Zhang Q, Ling M, Li X and Tan X: METTL14 promotes fibroblast-like synoviocytes activation via the LASP1/SRC/AKT axis in rheumatoid arthritis. Am J Physiol Cell Physiol. 324:C1089–C1100. 2023.PubMed/NCBI View Article : Google Scholar
32	Li H, Feng Y, Zheng X, Jia M, Mei Z, Wang Y, Zhang Z, Zhou M and Li C: M2-type exosomes nanoparticles for rheumatoid arthritis therapy via macrophage re-polarization. J Control Release. 341:16–30. 2022.PubMed/NCBI View Article : Google Scholar
33	Song Y, Gao N, Yang Z, Zhang L, Wang Y, Zhang S and Fan T: Characteristics, polarization and targeted therapy of mononuclear macrophages in rheumatoid arthritis. Am J Transl Res. 15:2109–2121. 2023.PubMed/NCBI
34	Gu Q, Yang H and Shi Q: Macrophages and bone inflammation. J Orthop Translat. 10:86–93. 2017.PubMed/NCBI View Article : Google Scholar
35	Li Y, Zhou Y, Wang Y, Crawford R and Xiao Y: Synovial macrophages in cartilage destruction and regeneration-lessons learnt from osteoarthritis and synovial chondromatosis. Biomed Mater. 17:2021.PubMed/NCBI View Article : Google Scholar
36	Huang X, Wang L, Guo H and Zhang W: Macrophage membrane-coated nanovesicles for dual-targeted drug delivery to inhibit tumor and induce macrophage polarization. Bioact Mater. 23:69–79. 2023.PubMed/NCBI View Article : Google Scholar
37	Zheng Y, Li Y, Ran X, Wang D, Zheng X, Zhang M, Yu B, Sun Y and Wu J: Mettl14 mediates the inflammatory response of macrophages in atherosclerosis through the NF-κB/IL-6 signaling pathway. Cell Mol Life Sci. 79(311)2022.PubMed/NCBI View Article : Google Scholar
38	Xie Q, Li Z, Luo X, Wang D, Zhou Y, Zhao J, Gao S, Yang Y, Fu W, Kong L and Sun T: piRNA-14633 promotes cervical cancer cell malignancy in a METTL14-dependent m6A RNA methylation manner. J Transl Med. 20(51)2022.PubMed/NCBI View Article : Google Scholar
39	Li T, Wang T, Jing J and Sun L: Expression pattern and clinical value of key m6A RNA modification regulators in abdominal aortic aneurysm. J Inflamm Res. 14:4245–4258. 2021.PubMed/NCBI View Article : Google Scholar
40	Fang Q, Zhou C and Nandakumar KS: Molecular and cellular pathways contributing to joint damage in rheumatoid arthritis. Mediators Inflamm. 2020(3830212)2020.PubMed/NCBI View Article : Google Scholar
41	Neamatallah T: Mitogen-activated protein kinase pathway: A critical regulator in tumor-associated macrophage polarization. J Microsc Ultrastruct. 7:53–56. 2019.PubMed/NCBI View Article : Google Scholar
42	Hao J, Hu Y, Li Y, Zhou Q and Lv X: Involvement of JNK signaling in IL4-induced M2 macrophage polarization. Exp Cell Res. 357:155–162. 2017.PubMed/NCBI View Article : Google Scholar
43	Wang Y, Han CC, Cui D, Li Y, Ma Y and Wei W: Is macrophage polarization important in rheumatoid arthritis? Int Immunopharmacol. 50:345–352. 2017.PubMed/NCBI View Article : Google Scholar