Department of Pediatrics/Key Laboratory of Birth Defects and Related Diseases of Women and Children (Ministry of Education), West China Second University Hospital, Sichuan University, Chengdu, 610041, China.
West China College of Stomatology/State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, 610041, China.
Hum Genomics. 2023 Aug 25;17(1):78. doi: 10.1186/s40246-023-00527-y.
The RNA m6A modification has been implicated in multiple neurological diseases as well as macrophage activation. However, whether it regulates microglial activation during hypoxic-ischemic brain damage (HIBD) in neonates remains unknown. Here, we aim to examine whether the m6A modification is involved in modulating microglial activation during HIBD. We employed an oxygen and glucose deprivation microglial model for in vitro studies and a neonatal mouse model of HIBD. The brain tissue was subjected to RNA-seq to screen for significant changes in the mRNA m6A regulator. Thereafter, we performed validation and bioinformatics analysis of the major m6A regulators.
RNA-seq analysis revealed that, among 141 m6A regulators, 31 exhibited significant differential expression (FC (abs) ≥ 2) in HIBD mice. We then subjected the major m6A regulators Mettl3, Mettl14, Fto, Alkbh5, Ythdf1, and Ythdf2 to further validation, and the results showed that all were significantly downregulated in vitro and in vivo. GO analysis reveals that regulators are mainly involved in the regulation of cellular and metabolic processes. The KEGG results indicate the involvement of the signal transduction pathway.
Our findings demonstrate that m6A modification of mRNA plays a crucial role in the regulation of microglial activation in HIBD, with m6A-associated regulators acting as key modulators of microglial activation.
RNA m6A 修饰与多种神经疾病以及巨噬细胞激活有关。然而，它是否调节新生儿缺氧缺血性脑损伤（HIBD）期间的小胶质细胞激活尚不清楚。在这里，我们旨在研究 m6A 修饰是否参与调节 HIBD 期间的小胶质细胞激活。我们使用氧葡萄糖剥夺小胶质细胞模型进行体外研究和新生鼠 HIBD 模型。对脑组织进行 RNA-seq 筛选以筛选出 mRNA m6A 调节剂的显著变化。此后，我们对主要 m6A 调节剂进行了验证和生物信息学分析。
RNA-seq 分析显示，在 141 个 m6A 调节剂中，有 31 个在 HIBD 小鼠中表现出显著的差异表达（FC（abs）≥2）。然后，我们对主要的 m6A 调节剂 Mettl3、Mettl14、Fto、Alkbh5、Ythdf1 和 Ythdf2 进行了进一步验证，结果表明它们在体外和体内均显著下调。GO 分析显示，调节剂主要参与细胞和代谢过程的调节。KEGG 结果表明信号转导途径的参与。
我们的研究结果表明，mRNA 的 m6A 修饰在 HIBD 中小胶质细胞激活的调节中起着至关重要的作用，m6A 相关调节剂作为小胶质细胞激活的关键调节剂。
