Yang Chuanxi, Zhao Kun, Zhang Jing, Wu Xiaoguang, Sun Wei, Kong Xiangqing, Shi Jing
Department of Cardiology, Medical School of Southeast University, Nanjing, China.
Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Front Cardiovasc Med. 2021 Mar 22;8:633631. doi: 10.3389/fcvm.2021.633631. eCollection 2021.
To systematically classify the profile of the RNA m6A modification landscape of neonatal heart regeneration. Cardiomyocyte proliferation markers were detected via immunostaining. The expression of m6A modification regulators was detected using quantitative real-time PCR (qPCR) and Western blotting. Genome-wide profiling of methylation-modified transcripts was conducted with methylation-modified RNA immunoprecipitation sequencing (m6A-RIP-seq) and RNA sequencing (RNA-seq). The Gene Expression Omnibus database (GEO) dataset was used to verify the hub genes. METTL3 and the level of m6A modification in total RNA was lower in P7 rat hearts than in P0 ones. In all, 1,637 methylation peaks were differentially expressed using m6A-RIP-seq, with 84 upregulated and 1,553 downregulated. Furthermore, conjoint analyses of m6A-RIP-seq, RNA-seq, and GEO data generated eight potential hub genes with differentially expressed hypermethylated or hypomethylated m6A levels. Our data provided novel information on m6A modification changes between Day 0 and Day 7 cardiomyocytes, which identified that increased METTL3 expression may enhance the proliferative capacity of neonatal cardiomyocytes, providing a theoretical basis for future clinical studies on the direct regulation of m6A in the proliferative capacity of cardiomyocytes.
为了系统地分类新生儿心脏再生过程中RNA m6A修饰图谱的特征。通过免疫染色检测心肌细胞增殖标志物。使用定量实时PCR（qPCR）和蛋白质印迹法检测m6A修饰调节因子的表达。利用甲基化修饰的RNA免疫沉淀测序（m6A-RIP-seq）和RNA测序（RNA-seq）对甲基化修饰的转录本进行全基因组分析。使用基因表达综合数据库（GEO）数据集验证核心基因。与P0大鼠心脏相比，P7大鼠心脏中METTL3和总RNA中的m6A修饰水平较低。使用m6A-RIP-seq共检测到1637个甲基化峰差异表达，其中84个上调，1553个下调。此外，对m6A-RIP-seq、RNA-seq和GEO数据的联合分析产生了8个潜在的核心基因，其m6A水平存在差异表达的高甲基化或低甲基化。我们的数据提供了有关第0天和第7天心肌细胞之间m6A修饰变化的新信息，确定了METTL3表达增加可能增强新生儿心肌细胞的增殖能力，为未来关于m6A对心肌细胞增殖能力直接调节的临床研究提供了理论基础。
