Transcriptome-Wide N6-Methyladenosine Alternations in Pulmonary Arteries of Monocrotaline-Induced Pulmonary Arterial Hypertension in Rats and Novel Therapeutic Targets

Yilu Feng; Zaixin Yu; Mi Tang; Jiang Li; Baohua Peng; Mukamengjiang Juaiti; Yiyang Tang; Benhui Liang; Mingqi Ouyang; Qingqing Liu; Jie Song

doi:10.3390/biomedicines12020364

Transcriptome-Wide N6-Methyladenosine Alternations in Pulmonary Arteries of Monocrotaline-Induced Pulmonary Arterial Hypertension in Rats and Novel Therapeutic Targets

Biomedicines. 2024 Feb 4;12(2):364. doi: 10.3390/biomedicines12020364.

Authors

Yilu Feng^{1

2

3}, Zaixin Yu¹, Mi Tang⁴, Jiang Li^{2

3}, Baohua Peng¹, Mukamengjiang Juaiti¹, Yiyang Tang¹, Benhui Liang¹, Mingqi Ouyang^{2

3}, Qingqing Liu⁵, Jie Song^{2

3}

Affiliations

¹ Department of Cardiology, Xiangya Hospital, Central South University, Changsha 410008, China.
² Research Institute of Blood Lipid and Atherosclerosis, Central South University, Changsha 410011, China.
³ Department of Cardiovascular Medicine, The Second Xiangya Hospital, Central South University, Changsha 410011, China.
⁴ Department of Cardiovascular Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China.
⁵ Department of Respiratory and Critical Care, The Second Xiangya Hospital, Central South University, Changsha 410011, China.

Abstract

N6-methyladenosine (m⁶A) is a post-transcriptional epigenetic change with transcriptional stability and functionality regulated by specific m⁶A-modifying enzymes. However, the significance of genes modified by m⁶A and enzymes specific to m⁶A regulation in the context of pulmonary arterial hypertension (PAH) remains largely unexplored. MeRIP-seq and RNA-seq were applied to explore variances in m⁶A 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 m⁶A-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 m⁶A 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 m⁶A. These genes are predominantly related to the extracellular matrix structure, MAPK, and PI3K/AKT pathways. A candidate gene, centromere protein F (CENPF), was detected with increased expression in the PAH group. Additionally, we first identified an m⁶A reader, leucine rich pentatricopeptide repeat containing (LRPPRC), which was downregulated in the PAH rat model. The in vitro downregulation of Lrpprc mediated by siRNA resulted in the enhanced proliferation and elevated expression of Cenpf mRNA in primary rat PASMCs. Our study revealed a modified transcriptome-wide m⁶A landscape and associated regulatory mechanisms in the pulmonary arteries of PAH rats, potentially offering a novel target for therapeutic strategies in the future.

Keywords: N6-methyladenosine; centromere protein F; leucine rich pentatricopeptide repeat containing; pulmonary arterial hypertension; pulmonary artery smooth muscle cells.

Abstract

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