Epitranscriptomic Analysis of m6A Methylome After Peripheral Nerve Injury

Abstract

N6-methyladenosine (m6A) is one of the most plentiful internal RNA modifications, especially in eukaryotic messenger RNA (mRNA), which plays pivotal roles in the regulation of mRNA life cycle and nerve development. However, the mRNA m6A methylation pattern in peripheral nervous injury (PNI) has not been investigated. In this study, sciatic nerve samples were collected from 7 days after sciatic nerve injury (SNI) and control rats. Quantitative real-time PCR demonstrated that m6A-related methyltransferase/demethylase genes were remarkably upregulated in SNI group compared with control group. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was performed to reveal the m6A methylation landscape. The results showed that 4,014 m6A peaks were significantly altered, including 2,144 upregulated and 1,870 downregulated m6A peaks, which were corresponded to 1,858 genes. Moreover, 919 differentially expressed genes were identified by the conjoint analysis of MeRIP-seq and RNA-seq. GO and KEGG pathway analyses were performed to determine the biological functions and signaling pathways of the m6A-modified genes. Notably, these genes were mainly related to the immune system process, cell activation, and nervous system development in GO analysis. KEGG pathway analysis revealed that these genes were involved in the cell cycle, B cell receptor signaling pathway, axon guidance pathway, and calcium signaling pathway. Furthermore, the m6A methylation and protein expression levels of autophagy-related gene (Atg7) were increased, together with the activation of autophagy. These findings shed some light on the epigenetic regulation of gene expression, which may provide a new opinion to promote functional recovery after PNI.

Alteration of m6A epitranscriptomic tagging of ribonucleic acids after spinal cord injury in mice

“…Our previous study demonstrated that the epigenetic network is essential for vascular regeneration and functional recovery post-SCI ( Ni et al, 2019 ). The m 6 A methylation plays a significant role in the pathological process of corneal injury, brain injury, and peripheral nerve injury ( Weng et al, 2018 ; Wang et al, 2019 ; Dai et al, 2021 ; Zhang et al, 2021 ). In contrast, various types of cancers and diabetes are associated with lowered m 6 A abundance ( Chen et al, 2020 ; Huang et al, 2021 ; Lei et al, 2021 ; Ruan et al, 2021 ; Wang et al, 2021a ; Ye et al, 2021 ).…”

RNA m6A Modification in Immunocytes and DNA Repair: The Biological Functions and Prospects in Clinical Application

“…Last but not least, m 6 A modification is supposed to be highly relevant to gut microbiota (Jabs et al, 2020), heat shock proteins (Feng et al, 2020), sepsis (Sun et al, 2020;Xing et al, 2021), and pulmonary hypertension (Pan et al, 2020) and even peripheral nerve injury (Zhang L et al, 2021). These studies can provide valuable experimental basis for development of new treatments.…”

Crosstalk among N6-methyladenosine modification and RNAs in central nervous system injuries

“…After nerve injury, m6A modification may have the function of regulating axonal regeneration and synaptic regeneration. Zhang et al (2021b) performed MeRIP-seq to reveal the m6A methylation landscape in peripheral nervous injury (PNI). They found that 4,014 m6A peaks were significantly altered after PNI.…”

“…They found that 4,014 m6A peaks were significantly altered after PNI. Moreover, GO analysis and KEGG pathway analysis showed that these genes were mainly involved in axon regeneration ( Zhang et al, 2021b ). Furthermore, Wang X. L. et al (2022) reported that in a mouse neuropathic pain (NP) model, downregulation of FTO in the anterior cingulate cortex (ACC) could promote angiogenesis, axon growth and neural plasticity as proven by up-regulation of matrix metalloproteinase-9 (MMP-9), decreased levels of precursor brain-derived neurotrophic factor (proBDNF) and increased levels of mature brain-derived neurotrophic factor (mBDNF).…”