Mettl3‑mediated MA RNA Methylation Regulates Osteolysis Induced by Titanium Particles

Overview

Journal Mol Med Rep

Specialty Molecular Biology

Date 2024 Jan 12

PMID 38214327

Authors

Xiaoxuan Lin

Yang Yang

Yaohong Huang

E Li

Xiumei Zhuang

Zhengchuan Zhang

Ruogu Xu

Xiaolin Yu

Feilong Deng

Affiliations

Soon will be listed here.

Abstract

Peri‑prosthetic osteolysis (PPO) induced by wear particles is considered the primary cause of titanium prosthesis failure and revision surgery. The specific molecular mechanisms involve titanium particles inducing multiple intracellular pathways, which impact disease prevention and the targeted therapy of PPO. Notably, N6‑methyladenosine (mA) serves critical roles in epigenetic regulation, particularly in bone metabolism and inflammatory responses. Thus, the present study aimed to determine the role of RNA methylation in titanium particle‑induced osteolysis. Results of reverse transcription‑quantitative PCR (RT‑qPCR), western blotting, ELISA and RNA dot blot assays revealed that titanium particles induced osteogenic inhibition and proinflammatory responses, accompanied by the reduced expression of methyltransferase‑like (Mettl) 3, a key component of m6A methyltransferase. Specific lentiviruses vectors were employed for Mettl3 knockdown and overexpression experiments. RT‑qPCR, western blotting and ELISA revealed that the knockdown of Mettl3 induced osteogenic inhibition and proinflammatory responses comparable with that induced by titanium particle, while Mettl3 overexpression attenuated titanium particle‑induced cellular reactions. Methylated RNA immunoprecipitation‑qPCR results revealed that titanium particles mediated the methylation of two inhibitory molecules, namely Smad7 and SMAD specific E3 ubiquitin protein ligase 1, via Mettl3 in bone morphogenetic protein signaling, leading to osteogenic inhibition. Furthermore, titanium particles induced activation of the nucleotide binding oligomerization domain 1 signaling pathway through methylation regulation, and the subsequent activation of the MAPK and NF‑κB pathways. Collectively, the results of the present study indicated that titanium particles utilized Mettl3 as an upstream regulatory molecule to induce osteogenic inhibition and inflammatory responses. Thus, the present study may provide novel insights into potential therapeutic targets for aseptic loosening in titanium prostheses.

References

Wu M, Chen G, Li Y . TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Res. 2016; 4:16009. PMC: 4985055. DOI: 10.1038/boneres.2016.9. View

Derks J, Tomasi C . Peri-implant health and disease. A systematic review of current epidemiology. J Clin Periodontol. 2014; 42 Suppl 16:S158-71. DOI: 10.1111/jcpe.12334. View

Zhou J, Wan J, Gao X, Zhang X, Jaffrey S, Qian S . Dynamic m(6)A mRNA methylation directs translational control of heat shock response. Nature. 2015; 526(7574):591-4. PMC: 4851248. DOI: 10.1038/nature15377. View

Zhu Z, Xie Q, Huang Y, Zhang S, Chen Y . Aucubin suppresses Titanium particles‑mediated apoptosis of MC3T3‑E1 cells and facilitates osteogenesis by affecting the BMP2/Smads/RunX2 signaling pathway. Mol Med Rep. 2018; 18(3):2561-2570. PMC: 6102688. DOI: 10.3892/mmr.2018.9286. View

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

Song H, Song J, Cheng M, Zheng M, Wang T, Tian S . METTL3-mediated mA RNA methylation promotes the anti-tumour immunity of natural killer cells. Nat Commun. 2021; 12(1):5522. PMC: 8448775. DOI: 10.1038/s41467-021-25803-0. View

Luo S, Liao C, Zhang L, Ling C, Zhang X, Xie P . METTL3-mediated m6A mRNA methylation regulates neutrophil activation through targeting TLR4 signaling. Cell Rep. 2023; 42(3):112259. DOI: 10.1016/j.celrep.2023.112259. View

Ollivere B, Wimhurst J, Clark I, Donell S . Current concepts in osteolysis. J Bone Joint Surg Br. 2012; 94(1):10-5. DOI: 10.1302/0301-620X.94B1.28047. View

Choi M, Koh H, Kluess D, OConnor D, Mathur A, Truskey G . Effects of titanium particle size on osteoblast functions in vitro and in vivo. Proc Natl Acad Sci U S A. 2005; 102(12):4578-83. PMC: 555523. DOI: 10.1073/pnas.0500693102. View

10.

Zhang Y, Gu X, Li D, Cai L, Xu Q . METTL3 Regulates Osteoblast Differentiation and Inflammatory Response via Smad Signaling and MAPK Signaling. Int J Mol Sci. 2020; 21(1). PMC: 6981640. DOI: 10.3390/ijms21010199. View

11.

Hsu P, Zhu Y, Ma H, Guo Y, Shi X, Liu Y . Ythdc2 is an N-methyladenosine binding protein that regulates mammalian spermatogenesis. Cell Res. 2017; 27(9):1115-1127. PMC: 5587856. DOI: 10.1038/cr.2017.99. View

12.

Zeng C, Huang W, Li Y, Weng H . Roles of METTL3 in cancer: mechanisms and therapeutic targeting. J Hematol Oncol. 2020; 13(1):117. PMC: 7457244. DOI: 10.1186/s13045-020-00951-w. View

13.

Delanois R, Mistry J, Gwam C, Mohamed N, Choksi U, Mont M . Current Epidemiology of Revision Total Knee Arthroplasty in the United States. J Arthroplasty. 2017; 32(9):2663-2668. DOI: 10.1016/j.arth.2017.03.066. View

14.

Shah R, Penmetsa D, Thomas R, Mehta D . Titanium Corrosion: Implications For Dental Implants. Eur J Prosthodont Restor Dent. 2017; 24(4):171-180. DOI: 10.1922/EJPRD_1531Shah10. View

15.

Yan X, Liu Z, Chen Y . Regulation of TGF-beta signaling by Smad7. Acta Biochim Biophys Sin (Shanghai). 2009; 41(4):263-72. PMC: 7110000. DOI: 10.1093/abbs/gmp018. View

16.

Caruso R, Warner N, Inohara N, Nunez G . NOD1 and NOD2: signaling, host defense, and inflammatory disease. Immunity. 2014; 41(6):898-908. PMC: 4272446. DOI: 10.1016/j.immuni.2014.12.010. View

17.

Wang P, Doxtader K, Nam Y . Structural Basis for Cooperative Function of Mettl3 and Mettl14 Methyltransferases. Mol Cell. 2016; 63(2):306-317. PMC: 4958592. DOI: 10.1016/j.molcel.2016.05.041. View

18.

Lin Z, Hsu P, Xing X, Fang J, Lu Z, Zou Q . Mettl3-/Mettl14-mediated mRNA N-methyladenosine modulates murine spermatogenesis. Cell Res. 2017; 27(10):1216-1230. PMC: 5630681. DOI: 10.1038/cr.2017.117. View

19.

Tsuchiya K, Yoshimura K, Inoue Y, Iwashita Y, Yamada H, Kawase A . YTHDF1 and YTHDF2 are associated with better patient survival and an inflamed tumor-immune microenvironment in non-small-cell lung cancer. Oncoimmunology. 2021; 10(1):1962656. PMC: 8366544. DOI: 10.1080/2162402X.2021.1962656. View

20.

Geng D, Wu J, Shao H, Zhu S, Wang Y, Zhang W . Pharmaceutical inhibition of glycogen synthetase kinase 3 beta suppresses wear debris-induced osteolysis. Biomaterials. 2015; 69:12-21. DOI: 10.1016/j.biomaterials.2015.07.061. View