Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-Innovation Center of Neuroregeneration, NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China.
Department of Neurosurgery, Binhai County People's Hospital, Yancheng, 224500, Jiangsu, People's Republic of China.
J Transl Med. 2023 Nov 23;21(1):845. doi: 10.1186/s12967-023-04694-3.
Denervation-induced muscle atrophy is complex disease involving multiple biological processes with unknown mechanisms. N6-methyladenosine (m6A) participates in skeletal muscle physiology by regulating multiple levels of RNA metabolism, but its impact on denervation-induced muscle atrophy is still unclear. Here, we aimed to explore the changes, functions, and molecular mechanisms of m6A RNA methylation during denervation-induced muscle atrophy.
During denervation-induced muscle atrophy, the m6A immunoprecipitation sequencing (MeRIP-seq) as well as enzyme-linked immunosorbent assay analysis were used to detect the changes of m6A modified RNAs and the involved biological processes. 3-deazidenosine (Daa) and R-2-hydroxyglutarate (R-2HG) were used to verify the roles of m6A RNA methylation. Through bioinformatics analysis combined with experimental verification, the regulatory roles and mechanisms of m6A RNA methylation had been explored.
There were many m6A modified RNAs with differences during denervation-induced muscle atrophy, and overall, they were mainly downregulated. After 72 h of denervation, the biological processes involved in the altered mRNA with m6A modification were mainly related to zinc ion binding, ubiquitin protein ligase activity, ATP binding and sequence-specific DNA binding and transcription coactivator activity. Daa reduced overall m6A levels in healthy skeletal muscles, which reduced skeletal muscle mass. On the contrary, the increase in m6A levels mediated by R-2HG alleviated denervation induced muscle atrophy. The m6A RNA methylation regulated skeletal muscle mass through ubiquitin-proteasome pathway.
This study indicated that decrease in m6A RNA methylation was a new symptom of denervation-induced muscle atrophy, and confirmed that targeting m6A alleviated denervation-induced muscle atrophy.
去神经诱导的肌肉萎缩是一种涉及多种未知机制的复杂疾病。N6-甲基腺苷（m6A）通过调节 RNA 代谢的多个水平参与骨骼肌生理学，但它对去神经诱导的肌肉萎缩的影响尚不清楚。在这里，我们旨在探讨 m6A RNA 甲基化在去神经诱导的肌肉萎缩过程中的变化、功能和分子机制。
在去神经诱导的肌肉萎缩过程中，使用 m6A 免疫沉淀测序（MeRIP-seq）以及酶联免疫吸附分析来检测 m6A 修饰 RNA 的变化及其涉及的生物过程。3-去氮腺苷（Daa）和 R-2-羟基戊二酸（R-2HG）用于验证 m6A RNA 甲基化的作用。通过生物信息学分析结合实验验证，探讨了 m6A RNA 甲基化的调控作用和机制。
在去神经诱导的肌肉萎缩过程中，有许多 m6A 修饰的 RNA 存在差异，总体上主要下调。去神经后 72 小时，m6A 修饰改变的 mRNA 涉及的生物过程主要与锌离子结合、泛素蛋白连接酶活性、ATP 结合和序列特异性 DNA 结合以及转录共激活因子活性有关。Daa 降低了健康骨骼肌中的整体 m6A 水平，从而减少了骨骼肌质量。相反，R-2HG 介导的 m6A 水平增加缓解了去神经诱导的肌肉萎缩。m6A RNA 甲基化通过泛素-蛋白酶体途径调节骨骼肌质量。
本研究表明，m6A RNA 甲基化减少是去神经诱导的肌肉萎缩的一个新症状，并证实靶向 m6A 可缓解去神经诱导的肌肉萎缩。
