RNA N6-methyladenosine modulates endothelial atherogenic responses to disturbed flow in mice

  1. Bochuan Li
  2. Ting Zhang
  3. Mengxia Liu
  4. Zhen Cui
  5. Yanhong Zhang
  6. Mingming Liu
  7. Yanan Liu
  8. Yongqiao Sun
  9. Mengqi Li
  10. Yikui Tian
  11. Ying Yang  Is a corresponding author
  12. Hongfeng Jiang  Is a corresponding author
  13. Degang Liang  Is a corresponding author
  1. Tianjin Medical University, China
  2. Chinese Academy of Sciences, China
  3. Capital Medical University, China

Abstract

Atherosclerosis preferentially occurs in atheroprone vasculature where human umbilical vein endothelial cells (HUVECs) are exposed to disturbed flow. Disturbed flow is associated with vascular inflammation and focal distribution. Recent studies have revealed the involvement of epigenetic regulation in atherosclerosis progression. N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic mRNA, but its function in endothelial atherogenic progression remains unclear. Here, we show that m6A mediates the EGFR signaling pathway during EC activation to regulate the atherosclerotic process. Oscillatory stress (OS) reduced the expression of METTL3, the primary m6A methyltransferase. Through m6A sequencing and functional studies, we determined that m6A mediates the mRNA decay of the vascular pathophysiology gene EGFR which leads to EC dysfunction. m6A modification of the EGFR 3'UTR accelerated its mRNA degradation. Double mutation of the EGFR 3'UTR abolished METTL3-induced luciferase activity. Adenovirus-mediated METTL3 overexpression significantly reduced EGFR activation and endothelial dysfunction in the presence of OS. Furthermore, TSP-1, an EGFR ligand, was specifically expressed in atheroprone regions without being affected by METTL3. Inhibition of the TSP-1/EGFR axis by using shRNA and AG1478 significantly ameliorated atherogenesis. Overall, our study revealed that METTL3 alleviates endothelial atherogenic progression through m6A-dependent stabilization of EGFR mRNA, highlighting the important role of RNA transcriptomics in atherosclerosis regulation.

Data availability

RNA-seq and MeRIP-seq data generated in this study have been deposited to the Genome Sequence Archive in BIG Data Center under accession number PRJCA004746.

The following data sets were generated

Article and author information

Author details

  1. Bochuan Li

    Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Ting Zhang

    CAS Key Laboratory of Genomic and Precision Medicine, Chinese Academy of Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Mengxia Liu

    CAS Key Laboratory of Genomic and Precision Medicine, Chinese Academy of Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Zhen Cui

    Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Yanhong Zhang

    Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Mingming Liu

    Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Yanan Liu

    Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Yongqiao Sun

    CAS Key Laboratory of Genomic and Precision Medicine, Chinese Academy of Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Mengqi Li

    Tianjin Medical University General Hospital Cardiovascular Department, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Yikui Tian

    Tianjin Medical University General Hospital Cardiovascular Department, Tianjin Medical University, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  11. Ying Yang

    CAS Key Laboratory of Genomic and Precision Medicine, Chinese Academy of Sciences, Beijing, China
    For correspondence
    yingyang@big.ac.cn
    Competing interests
    The authors declare that no competing interests exist.
  12. Hongfeng Jiang

    Key Laboratory of Remodeling-Related Cardiovascular Diseases, Capital Medical University, Tianjin, China
    For correspondence
    jhf@pku.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
  13. Degang Liang

    Tianjin Medical University General Hospital Cardiovascular Department, Tianjin Medical University, Tianjin, China
    For correspondence
    15922230066@163.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2618-6651

Funding

National Natural Science Foundation of China (81900396)

  • Bochuan Li

National Natural Science Foundation of China (82000477)

  • Mengqi Li

National Natural Science Foundation of China (81970392)

  • Hongfeng Jiang

Postdoctoral Research Foundation of China (2019M661041)

  • Bochuan Li

Postdoctoral Research Foundation of China (BX20190235)

  • Bochuan Li

China Association for Science and Technology (Excellent Sino-foreign Youth Exchange Program)

  • Bochuan Li

National Natural Science Foundation of China (91940304)

  • Ying Yang

Chinese Academy of Sciences (2018133)

  • Ying Yang

National Key Research and Development Program of China (2018YFA0801200)

  • Ying Yang

Beijing Nova Program (Z201100006820104)

  • Ying Yang

National Natural Science Foundation of China (81870207)

  • Yikui Tian

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: The investigation conformed to the Guide for the Care and Use of Laboratory Animals by the US National Institutes of Health (NIH 17 Publication No. 85-23, revised in 2011). All study protocols and the use of animals were approved by the Institutional Animal Care and Use Committee of Tianjin Medical University.

Reviewing Editor

  1. Edward A Fisher, New York University Grossman School of Medicine, United States

Publication history

  1. Received: April 29, 2021
  2. Accepted: January 7, 2022
  3. Accepted Manuscript published: January 10, 2022 (version 1)
  4. Version of Record published: January 27, 2022 (version 2)

Copyright

© 2022, Li et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Bochuan Li
  2. Ting Zhang
  3. Mengxia Liu
  4. Zhen Cui
  5. Yanhong Zhang
  6. Mingming Liu
  7. Yanan Liu
  8. Yongqiao Sun
  9. Mengqi Li
  10. Yikui Tian
  11. Ying Yang
  12. Hongfeng Jiang
  13. Degang Liang
(2022)
RNA N6-methyladenosine modulates endothelial atherogenic responses to disturbed flow in mice
eLife 11:e69906.
https://doi.org/10.7554/eLife.69906

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