Exploration of N6-Methyladenosine Profiles of mRNAs and the Function of METTL3 in Atherosclerosis

Yaqing Zhou; Rongli Jiang; Yali Jiang; Yahong Fu; Yerbolat Manafhan; Jinfu Zhu; Enzhi Jia

doi:10.3390/cells11192980

Exploration of N6-Methyladenosine Profiles of mRNAs and the Function of METTL3 in Atherosclerosis

Cells. 2022 Sep 24;11(19):2980. doi: 10.3390/cells11192980.

Authors

Yaqing Zhou¹, Rongli Jiang¹, Yali Jiang², Yahong Fu¹, Yerbolat Manafhan³, Jinfu Zhu⁴, Enzhi Jia¹

Affiliations

¹ Department of Cardiovascular Medicine, The First Affiliated Hospital of Nanjing Medical University, Guangzhou Road 300, Nanjing 210029, China.
² The Friendship Hospital of Ili Kazakh Autonomous Prefecture，Ili & Jiangsu Joint Institute of Health, Yining 835000, China.
³ Department of Hypertension, Yili Friendship Hospital, Stalin Road 92, Yining 835000, China.
⁴ Department of Cardiovascular Surgery, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.

Abstract

Objectives: N6-methylladenosine (m6A) modification has not been fully studied in atherosclerosis. The objectives of this study were to investigate differentially expressed m6A methylated peaks and mRNAs, along with the regulatory role of methyltransferase 3 (METTL3) in pathological processes of atherosclerosis.

Methods: The pathological models of human coronary artery smooth muscle cells (HCASMCs) were induced in vitro. The differentially expressed mRNAs and m6A peaks were identified by RNA-Seq and meRIP-Seq. The potential mechanisms were analyzed via bioinformatic assays. Methylases expression was tested by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB) in HCASMCs, and by immunohistochemical assays in 40 human coronary arteries. The knockdown of METTL3 expression in cells was performed by siRNA transfection, and cell proliferation and migration were detected after transfection.

Results: We identified 5121 m6A peaks and 883 mRNAs that were expressed differentially in the pathological processes of HCASMCs. Bioinformatic analyses showed that the different m6A peaks were associated with cell growth and cell adhesion, and the 883 genes showed that the extracellular matrix and PI3K/AKT pathway regulate the processes of HCASMCs. Additionally, 10 hub genes and 351 mRNAs with differential methylation and expression levels were found. METTL3 was upregulated in the arteries with atherosclerotic lesions and in the proliferation and migration model of HCASMCs, and pathological processes of HCASMCs could be inhibited by the knockdown of METTL3. The mechanisms behind regulation of migration and proliferation reduced by siMETTL3 are concerned with protein synthesis and energy metabolism.

Conclusions: These results revealed a new m6A epigenetic method to regulate the progress of atherosclerosis, which suggest approaches for potential therapeutic interventions that target METTL3 for the prevention and treatment of coronary artery diseases.

Keywords: METTL3; RNA-Seq; atherosclerosis; meRIP-Seq; smooth muscle cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine / analogs & derivatives
Adenosine / metabolism
Atherosclerosis* / genetics
Cell Line, Tumor
Humans
Methyltransferases* / genetics
Methyltransferases* / metabolism
Phosphatidylinositol 3-Kinases
Proto-Oncogene Proteins c-akt
RNA, Messenger / genetics
RNA, Messenger / metabolism
RNA, Small Interfering

Substances

RNA, Messenger
RNA, Small Interfering
N-methyladenosine
Methyltransferases
METTL3 protein, human
Proto-Oncogene Proteins c-akt
Adenosine

Grants and funding

This study received support from the National Natural Science Foundation of China (No. 81970302) and the Science Foundation Project of the Yili and Jiangsu Joint Institute of Health (No. yl2021ms06, No. yl2020py01).