Feng Yilu, Yu Zaixin, Tang Mi, Li Jiang, Peng Baohua, Juaiti Mukamengjiang, Tang Yiyang, Liang Benhui, Ouyang Mingqi, Liu Qingqing, Song Jie
Department of Cardiology, Xiangya Hospital, Central South University, Changsha 410008, China.
Research Institute of Blood Lipid and Atherosclerosis, Central South University, Changsha 410011, China.
Biomedicines. 2024 Feb 4;12(2):364. doi: 10.3390/biomedicines12020364.
N6-methyladenosine (mA) is a post-transcriptional epigenetic change with transcriptional stability and functionality regulated by specific mA-modifying enzymes. However, the significance of genes modified by mA and enzymes specific to mA regulation in the context of pulmonary arterial hypertension (PAH) remains largely unexplored. MeRIP-seq and RNA-seq were applied to explore variances in mA and RNA expression within the pulmonary artery tissues of control and monocrotaline-induced PAH rats. Functional enrichments were analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. To screen candidate mA-related genes, the STRING and Metascape databases were used to construct a protein-protein interaction network followed by a real-time PCR validation of their expression. The expression level of an mA regulator was further investigated using immunohistochemical staining, immunofluorescence, and Western blot techniques. Additionally, proliferation assays were conducted on primary rat pulmonary artery smooth muscle cells (PASMCs). We identified forty-two differentially expressed genes that exhibited either hypermethylated or hypomethylated mA. These genes are predominantly related to the extracellular matrix structure, MAPK, and PI3K/AKT pathways. A candidate gene, centromere protein F (), was detected with increased expression in the PAH group. Additionally, we first identified an mA reader, leucine rich pentatricopeptide repeat containing (LRPPRC), which was downregulated in the PAH rat model. The in vitro downregulation of mediated by siRNA resulted in the enhanced proliferation and elevated expression of mRNA in primary rat PASMCs. Our study revealed a modified transcriptome-wide mA landscape and associated regulatory mechanisms in the pulmonary arteries of PAH rats, potentially offering a novel target for therapeutic strategies in the future.
N6-甲基腺苷（mA）是一种转录后表观遗传变化，其转录稳定性和功能由特定的mA修饰酶调节。然而，在肺动脉高压（PAH）背景下，由mA修饰的基因和mA调节特异性酶的意义仍 largely unexplored。应用MeRIP-seq和RNA-seq来探索对照和野百合碱诱导的PAH大鼠肺动脉组织中mA和RNA表达的差异。使用基因本体论和京都基因与基因组百科全书进行功能富集分析。为了筛选候选的mA相关基因，使用STRING和Metascape数据库构建蛋白质-蛋白质相互作用网络，随后对其表达进行实时PCR验证。使用免疫组织化学染色、免疫荧光和蛋白质印迹技术进一步研究mA调节剂的表达水平。此外，对原代大鼠肺动脉平滑肌细胞（PASMCs）进行增殖测定。我们鉴定出42个差异表达基因，这些基因表现出mA高甲基化或低甲基化。这些基因主要与细胞外基质结构、MAPK和PI3K/AKT途径相关。在PAH组中检测到一个候选基因，着丝粒蛋白F（）表达增加。此外，我们首次鉴定出一种mA阅读器，富含亮氨酸的五肽重复序列（LRPPRC），其在PAH大鼠模型中下调。由siRNA介导的体外下调导致原代大鼠PASMCs中增殖增强和mRNA表达升高。我们的研究揭示了PAH大鼠肺动脉中转录组范围内的mA修饰图谱及相关调控机制，可能为未来的治疗策略提供新靶点。
