Methotrexate attenuates vascular inflammation through an adenosine-microRNA dependent pathway

  1. Dafeng Yang
  2. Stefan Haemmig
  3. Haoyang Zhou
  4. Daniel Pérez-Cremades
  5. Xinghui Sun
  6. Lei Chen
  7. Jie Li
  8. Jorge Haneo-Mejia
  9. Tianlun Yang
  10. Ivana Hollan
  11. Mark W Feinberg  Is a corresponding author
  1. Brigham and Women's Hospital/Harvard Medical School, United States
  2. Central South University, China
  3. Xiangya Hospital, Central South University, China
  4. University of Pennsylvania, United States

Abstract

Endothelial cell (EC) activation is an early hallmark in the pathogenesis of chronic vascular diseases. MicroRNA-181b (MiR-181b) is an important anti-inflammatory mediator in the vascular endothelium affecting endotoxemia, atherosclerosis, and insulin resistance. Herein, we identify that the drug methotrexate (MTX) and its downstream metabolite adenosine exert anti-inflammatory effects in the vascular endothelium by targeting and activating MiR-181b expression. Both systemic and endothelial-specific MiR-181a2b2-deficient mice develop vascular inflammation, white adipose tissue (WAT) inflammation, and insulin resistance in a diet-induced obesity model. Moreover, MTX attenuated diet-induced WAT inflammation, insulin resistance, and EC activation in a MiR-181a2b2-dependent manner. Mechanistically, MTX attenuated cytokine-induced EC activation through a unique adenosine-adenosine receptor A3-SMAD3/4-MiR-181b signaling cascade. These findings establish an essential role of endothelial MiR-181b in controlling vascular inflammation and that restoring MiR-181b in ECs by high dose MTX or adenosine signaling may provide a potential therapeutic opportunity for anti-inflammatory therapy.

Data availability

Source data files have been provided for Figures 1 -2. RNA-Seq data has been made accessible.

Article and author information

Author details

  1. Dafeng Yang

    Medicine/Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Stefan Haemmig

    Medicine/Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Haoyang Zhou

    Cardiovascular, Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Daniel Pérez-Cremades

    Medicine, Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Xinghui Sun

    Department of Medicine, Cardiovascular Division, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Lei Chen

    Cardiology, Xiangya Hospital, Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Jie Li

    Medicine/Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Jorge Haneo-Mejia

    Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Tianlun Yang

    Cardiology, Xiangya Hospital, Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Ivana Hollan

    Medicine/Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Mark W Feinberg

    Medicine/Cardiology, Brigham and Women's Hospital/Harvard Medical School, Boston, United States
    For correspondence
    mfeinberg@bwh.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9523-3859

Funding

National Institutes of Health (HL115141)

  • Mark W Feinberg

National Institutes of Health (HL134849)

  • Mark W Feinberg

American Heart Association (18SFRN33900144)

  • Mark W Feinberg

American Heart Association (18POST34030395)

  • Stefan Haemmig

Falk Foundation

  • Mark W Feinberg

National Natural Science Foundation of China

  • Tianlun Yang

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

Ethics

Animal experimentation: All mice were maintained under SPF conditions at an American Association for the Accreditation of Laboratory Animal Care-accredited animal facility at the Brigham and Women's Hospital (protocol #2016N000182). All animal protocols were approved by the Institutional Animal Care and Use Committee at Harvard Medical School, Boston, MA and conducted in accordance with the National Institutes of Health Guide for Care and Use of Laboratory Animals.

Reviewing Editor

  1. Peter Tontonoz, University of California, Los Angeles, United States

Publication history

  1. Received: April 20, 2020
  2. Accepted: December 31, 2020
  3. Accepted Manuscript published: January 8, 2021 (version 1)
  4. Version of Record published: January 27, 2021 (version 2)

Copyright

© 2021, Yang 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.

Metrics

  • 1,147
    Page views
  • 126
    Downloads
  • 3
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Dafeng Yang
  2. Stefan Haemmig
  3. Haoyang Zhou
  4. Daniel Pérez-Cremades
  5. Xinghui Sun
  6. Lei Chen
  7. Jie Li
  8. Jorge Haneo-Mejia
  9. Tianlun Yang
  10. Ivana Hollan
  11. Mark W Feinberg
(2021)
Methotrexate attenuates vascular inflammation through an adenosine-microRNA dependent pathway
eLife 10:e58064.
https://doi.org/10.7554/eLife.58064

Further reading

    1. Immunology and Inflammation
    Manoj Arra et al.
    Research Article Updated

    Osteoarthritis is the most common joint disease in the world with significant societal consequences but lacks effective disease-modifying interventions. The pathophysiology consists of a prominent inflammatory component that can be targeted to prevent cartilage degradation and structural defects. Intracellular metabolism has emerged as a culprit of the inflammatory response in chondrocytes, with both processes co-regulating each other. The role of glutamine metabolism in chondrocytes, especially in the context of inflammation, lacks a thorough understanding and is the focus of this work. We display that mouse chondrocytes utilize glutamine for energy production and anabolic processes. Furthermore, we show that glutamine deprivation itself causes metabolic reprogramming and decreases the inflammatory response of chondrocytes through inhibition of NF-κB activity. Finally, we display that glutamine deprivation promotes autophagy and that ammonia is an inhibitor of autophagy. Overall, we identify a relationship between glutamine metabolism and inflammatory signaling and display the need for increased study of chondrocyte metabolic systems.

    1. Immunology and Inflammation
    Daniel Radtke et al.
    Research Article

    Th2 cells provide effector functions in type 2 immune responses to helminths and allergens. Despite knowledge about molecular mechanisms of Th2 cell differentiation, there is little information on Th2 cell heterogeneity and clonal distribution between organs. To address this, we performed combined single-cell transcriptome and TCR clonotype analysis on murine Th2 cells in mesenteric lymph nodes (MLN) and lung after infection with Nippostrongylus brasiliensis (Nb) as a human hookworm infection model. We find organ-specific expression profiles, but also populations with conserved migration or effector/resident memory signatures that unexpectedly cluster with potentially regulatory Il10posFoxp3neg cells. A substantial MLN subpopulation with an interferon response signature suggests a role for interferon-signaling in Th2 differentiation or diversification. Further RNA-inferred developmental directions indicate proliferation as a hub for differentiation decisions. Although the TCR repertoire is highly heterogeneous, we identified expanded clones and CDR3 motifs. Clonal relatedness between distant organs confirmed effective exchange of Th2 effector cells, although locally expanded clones dominated the response. We further cloned an Nb-specific TCR from an expanded clone in the lung effector cluster and describe surface markers that distinguish transcriptionally defined clusters. These results provide insights in Th2 cell subset diversity and clonal relatedness in distant organs.