METTL14-mediated m6A epitranscriptomic modification contributes to chemotherapy-induced neuropathic pain by stabilizing GluN2A expression via IGF2BP2
Weicheng Lu, … , Huijie Ma, Jingdun Xie
Weicheng Lu, … , Huijie Ma, Jingdun Xie
Published February 6, 2024
Citation Information: J Clin Invest. 2024;134(6):e174847. https://doi.org/10.1172/JCI174847.
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Research Article Neuroscience

METTL14-mediated m6A epitranscriptomic modification contributes to chemotherapy-induced neuropathic pain by stabilizing GluN2A expression via IGF2BP2

Abstract

Epigenetics is a biological process that modifies and regulates gene expression, affects neuronal function, and contributes to pain. However, the mechanism by which epigenetics facilitates and maintains chronic pain is poorly understood. We aimed to determine whether N6-methyladenosine (m6A) specifically modified by methyltransferase-like 14 (METTL14) alters neuronal activity and governs pain by sensitizing the GluN2A subunit of the N-methyl-d-aspartate receptor (NMDAR) in the dorsal root ganglion (DRG) neurons in a model of chemotherapy-induced neuropathic pain (CINP). Using dot blotting, immunofluorescence, gain/loss-of-function, and behavioral assays, we found that m6A levels were upregulated in L4–L6 DRG neurons in CINP in a DBP/METTL14-dependent manner, which was also confirmed in human DRGs. Blocking METTL14 reduced m6A methylation and attenuated pain hypersensitivity. Mechanistically, METTL14-mediated m6A modification facilitated the synaptic plasticity of DRG neurons by enhancing the GluN2A subunit of NMDAR, and inhibiting METTL14 blocked this effect. In contrast, overexpression of METTL14 upregulated m6A modifications, enhanced presynaptic NMDAR activity in DRG neurons, and facilitated pain sensation. Our findings reveal a previously unrecognized mechanism of METTL14-mediated m6A modification in DRG neurons to maintain neuropathic pain. Targeting these molecules may provide a new strategy for pain treatment.

Authors

Weicheng Lu, Xiaohua Yang, Weiqiang Zhong, Guojun Chen, Xinqi Guo, Qingqing Ye, Yixin Xu, Zhenhua Qi, Yaqi Ye, Jingyun Zhang, Yuge Wang, Xintong Wang, Shu Wang, Qiyue Zhao, Weian Zeng, Junting Huang, Huijie Ma, Jingdun Xie

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Figure 10

The transcription factor DBP mediates METTL14 upregulation caused by paclitaxel treatment.

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The transcription factor DBP mediates METTL14 upregulation caused by pac...
(A) Venn diagram shows the predictive transcription factors (TFs) by taking intersections with the TFs of the rat species and predictive results from the PROMO, AnimalTFDB, and MEME (FIMO function) databases. (B and C) Dbp mRNA (B) and protein (C) expression in the bilateral L4–L6 DRGs in saline- or PCX-pretreated rats (at least 6 rats per group, Student’s t test). (D) Colocalization analysis of DBP and METTL14 based on double-labeled immunofluorescent images. Scale bars: 100 μm, left; 50 μm, enlarged image. (E) DBP and METTL14 protein expression in PC12 cells transfected with siDbp compared with the control (n = 6 per group, 1-way ANOVA followed by Tukey’s post hoc test). (F) Dbp and Grin2a mRNA expression in the rat bilateral L4–L6 DRGs in saline- or PCX-treated rats with siRNA-Mettl14 or scrambled siRNA injection (n = 5 rats per group, 1-way ANOVA followed by Tukey’s post hoc test). (G–I) Mechanical allodynia, thermal hyperalgesia, and cold allodynia of saline- or PCX-treated rats after siRNA-Dbp or scrambled siRNA treatment (at least 7 rats per group, 2-way ANOVA followed by Tukey’s post hoc test). (J) Top: Schematic representation of the full-length Mettl14 promoter, truncation promoters (–780 bp to ~30 bp, –590 bp to ~30 bp), and predicted binding sites from the JASPAR database. Bottom: Vectors containing full-length Mettl14 promoter and Mettl14 truncation promoters (–780 bp to ~30 bp) show higher luciferase activity (at least n = 7 per group, 1-way ANOVA followed by Tukey’s post hoc test). Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.