Integrated analysis of m ⁶A mRNA methylation in rats with monocrotaline-induced pulmonary arterial hypertension

Abstract

Background: N6-methyladenosine (m ⁶A) modification is one of the most common chemical modifications of eukaryotic mRNAs, which play an important role in tumors and cardiovascular disease through regulating mRNA stability, splicing and translation. However, the changes of m ⁶A mRNA and m ⁶A-related enzymes in pulmonary arterial hypertension (PAH) remain largely unexplored.

Methods: MeRIP-seq was used to identify m ⁶A methylation in lung tissues from control and MCT-PAH rats. Western blot and immunofluorescence were used to evaluate expression of m ⁶A-related enzymes.

Results: Compared with control group, m ⁶A methylation was mainly increased in lung tissues from MCT-PAH rats. The up-methylated coding genes in MCT-PAH rats were primarily enriched in processes associated with inflammation, glycolysis, ECM-receptor interaction and PDGF signal pathway, while genes with down-methylation were enriched in processes associated with TGF-β family receptor members. The expression of FTO and ALKBH5 downregulated, METTL3 and YTHDF1 increased and other methylation modification-related proteins was not significantly changed in MCT-PAH rats lung tissues. Immunofluorescence indicated that expression of FTO decreased and YTHDF1 increased in small pulmonary arteries of MCT-PAH rats.

Conclusion: m ⁶A levels and the expression of methylation-related enzymes were altered in PAH rats, in which FTO and YTHDF1 may play a crucial role in m ⁶A modification.