Department of Orthopaedics, Qilu Hospital, Shandong University Centre for Orthopaedics, Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong University, Jinan, China; Shandong University Centre for Orthopaedics, Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong University, Jinan, China.
Department of Orthopaedics, Tianjin Medical University General Hospital, International Science and Technology Cooperation Base of Spinal Cord Injury, Tianjin Key Laboratory of Spine and Spinal Cord, Tianjin, China.
Epigenetics. 2023 Dec;18(1):2181575. doi: 10.1080/15592294.2023.2181575.
Cerebral ischaemiareperfusion injury is an important pathological process in nervous system diseases during which neurons undergo oxygenglucose deprivation and reoxygenation (OGD/R) injury. No study has used epitranscriptomics to explore the characteristics and mechanism of injury. N6methyladenosine (m6A) is the most abundant epitranscriptomic RNA modification. However, little is known about m6A modifications in neurons, especially during OGD/R. m6A RNA immunoprecipitation sequencing (MeRIPseq) and RNA-sequencing data for normal and OGD/R-treated neurons were analysed by bioinformatics. MeRIP quantitative real-time polymerase chain reaction was used to determine the m6A modification levels on specific RNAs. We report the m6A modification profiles of the mRNA and circRNA transcriptomes of normal and OGD/R-treated neurons. Expression analysis revealed that the m6A levels did not affect m6A mRNA or m6A circRNA expression. We found crosstalk between m6A mRNAs and m6A circRNAs and identified three patterns of m6A circRNA production in neurons; thus, distinct OGD/R treatments induced the same genes to generate different m6A circRNAs. Additionally, m6A circRNA biogenesis during distinct OGD/R processes was found to be time specific. These results expand our understanding of m6A modifications in normal and OGD/R-treated neurons, providing a reference to explore epigenetic mechanisms and potential treatments for OGD/R-related diseases.
脑缺血再灌注损伤是神经系统疾病中的一个重要病理过程，在此过程中神经元经历氧葡萄糖剥夺和再氧合（OGD/R）损伤。目前尚无研究使用表观转录组学来探索损伤的特征和机制。N6-甲基腺苷（m6A）是最丰富的表观转录组 RNA 修饰。然而，关于神经元中的 m6A 修饰，特别是在 OGD/R 期间，人们知之甚少。通过生物信息学分析正常和 OGD/R 处理神经元的 m6A RNA 免疫沉淀测序（MeRIPseq）和 RNA-seq 数据。使用 MeRIP 定量实时聚合酶链反应来确定特定 RNA 上的 m6A 修饰水平。我们报告了正常和 OGD/R 处理神经元的 mRNA 和 circRNA 转录组的 m6A 修饰谱。表达分析表明，m6A 水平不影响 m6A mRNA 或 m6A circRNA 的表达。我们发现 m6A mRNA 和 m6A circRNA 之间存在串扰，并在神经元中鉴定出三种 m6A circRNA 产生模式；因此，不同的 OGD/R 处理诱导相同的基因产生不同的 m6A circRNA。此外，在不同的 OGD/R 过程中 m6A circRNA 的生物发生被发现是时间特异性的。这些结果扩展了我们对正常和 OGD/R 处理神经元中 m6A 修饰的理解，为探索表观遗传机制和 OGD/R 相关疾病的潜在治疗方法提供了参考。
