METTL3 Affects Spinal Cord Neuronal Apoptosis by Regulating Bcl-2 m6A Modifications After Spinal Cord Injury

Abstract

Objective: Spinal cord injury (SCI) is a severe type of neurological trauma. N6-methyladenosine (m6A) modification is one of the most common internal modifications of RNA. The role of METTL3, the predominant methylation enzyme of m6A modification, in SCI remains unclear. This study aimed to investigate the role of methyltransferase METTL3 in SCI.Methods: After establishing the oxygen-glucose deprivation (OGD) model of PC12 cells and rat spinal cord hemisection model, we found that the expression of METTL3 and the overall m6A modification level were significantly increased in neurons. The m6A modification was identified on B-cell lymphoma 2 (Bcl-2) messenger RNA (mRNA) by bioinformatics analysis, and m6A-RNA immunoprecipitation and RNA immunoprecipitation. In addition, METTL3 was blocked by the specific inhibitor STM2457 and gene knockdown, and then apoptosis levels were measured.Results: In different models, we found that the expression of METTL3 and the overall m6A modification level were significantly increased in neurons. After inducing OGD, inhibition of METTL3 activity or expression increased the mRNA and protein levels of Bcl-2, inhibited neuronal apoptosis, and improved neuronal viability in the spinal cord.Conclusion: Inhibition of METTL3 activity or expression can inhibit the apoptosis of spinal cord neurons after SCI through the m6A/Bcl-2 signaling pathway.

Mechanism of Fat Mass and Obesity‐Related Gene‐Mediated Heme Oxygenase‐1 m6A Modification in the Recovery of Neurological Function in Mice with Spinal Cord Injury

“… 37 Additionally, m6A modification levels are heightened in neurons in the OGD model of PC12 cells. 11 FTO, an RNA demethylase, exhibits an inverse relationship with m6A levels. FTO knockout enhances m6A modification levels, while FTO overexpression decreases them.…”

“… 10 The m6A modification level exhibits a substantial increase in the oxygen/glucose deprivation (OGD) model in PC12 cells and SCI mice. 11 Fat mass and obesity‐associated protein (FTO) is the first reported RNA N6‐methyladenosine (m6A) demethylase in eukaryotic cells. 12 Silencing FTO elevates m6A modification levels, while FTO overexpression suppresses the m6A level.…”

“… 14 , 15 Additionally, the m6A modification level undergoes a conspicuous elevation in the rat spinal cord hemisection and PC12 cells in the OGD model. 11 Various reader proteins, including IGFBP, YTHDC, and YTHDF, selectively recognize m6A modifications and regulate mRNA stability. 15 Remarkably, YTHDF2 specifically binds to the m6A site, instigating the degradation of the complex by recruiting transcripts to the RNA.…”

METTL3 promotes the osteogenic differentiation of periosteum-derived MSCs via regulation of the HOXD8/ITGA5 axis in congenital pseudarthrosis

“…Furthermore, N 6 -methyladenosine (m 6 A) modification is engaged with the osteogenic differentiation regulation in MSCs [ 11 ]. And Methyltransferase-like 3 (METTL3) is an extensively explored catalytic subunit of methyltransferases complex in the m 6 A modification process [ 17 ]. The inhibition of METTL3 has restrained the RUNX2 expression and down-regulated the bone marrow stem cell's osteogenic differentiation capacity [ [18] , [19] , [20] , [21] ].…”