1. Immunology and Inflammation
  2. Medicine

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
https://doi.org/10.7554/eLife.58064
Share this article
Cite this article
  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
  2. Download BibTeX
  3. Download .RIS

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

  • 1,235
    Page views
  • 141
    Downloads
  • 5
    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

Categories and tags

Research organisms

    1. Of interest

    Glycine inhibits NINJ1 membrane clustering to suppress plasma membrane rupture in cell death

    Jazlyn P Borges, Ragnhild SR Sætra ... Benjamin Ethan Steinberg
  2. Further reading

Further reading

    1. Cell Biology
    2. Immunology and Inflammation

    Glycine inhibits NINJ1 membrane clustering to suppress plasma membrane rupture in cell death

    Jazlyn P Borges, Ragnhild SR Sætra ... Benjamin Ethan Steinberg
    Short Report

    First recognized more than 30 years ago, glycine protects cells against rupture from diverse types of injury. This robust and widely observed effect has been speculated to target a late downstream process common to multiple modes of tissue injury. The molecular target of glycine that mediates cytoprotection, however, remains elusive. Here, we show that glycine works at the level of NINJ1, a newly identified executioner of plasma membrane rupture in pyroptosis, necrosis, and post-apoptosis lysis. NINJ1 is thought to cluster within the plasma membrane to cause cell rupture. We demonstrate that the execution of pyroptotic cell rupture is similar for human and mouse NINJ1, and that NINJ1 knockout functionally and morphologically phenocopies glycine cytoprotection in macrophages undergoing lytic cell death. Next, we show that glycine prevents NINJ1 clustering by either direct or indirect mechanisms. In pyroptosis, glycine preserves cellular integrity but does not affect upstream inflammasome activities or accompanying energetic cell death. By positioning NINJ1 clustering as a glycine target, our data resolve a long-standing mechanism for glycine-mediated cytoprotection. This new understanding will inform the development of cell preservation strategies to counter pathologic lytic cell death.

    1. Chromosomes and Gene Expression
    2. Immunology and Inflammation

    SRSF6 balances mitochondrial-driven innate immune outcomes through alternative splicing of BAX

    Allison R Wagner, Chi G Weindel ... Kristin L Patrick
    Research Article Updated

    To mount a protective response to infection while preventing hyperinflammation, gene expression in innate immune cells must be tightly regulated. Despite the importance of pre-mRNA splicing in shaping the proteome, its role in balancing immune outcomes remains understudied. Transcriptomic analysis of murine macrophage cell lines identified Serine/Arginine Rich Splicing factor 6 (SRSF6) as a gatekeeper of mitochondrial homeostasis. SRSF6-dependent orchestration of mitochondrial health is directed in large part by alternative splicing of the pro-apoptosis pore-forming protein BAX. Loss of SRSF6 promotes accumulation of BAX-κ, a variant that sensitizes macrophages to undergo cell death and triggers upregulation of interferon stimulated genes through cGAS sensing of cytosolic mitochondrial DNA. Upon pathogen sensing, macrophages regulate SRSF6 expression to control the liberation of immunogenic mtDNA and adjust the threshold for entry into programmed cell death. This work defines BAX alternative splicing by SRSF6 as a critical node not only in mitochondrial homeostasis but also in the macrophage’s response to pathogens.

    1. Computational and Systems Biology
    2. Immunology and Inflammation

    Longitudinal analysis of invariant natural killer T cell activation reveals a cMAF-associated transcriptional state of NKT10 cells

    Harry Kane, Nelson M LaMarche ... Lydia Lynch
    Research Article

    Innate T cells, including CD1d-restricted invariant natural killer T (iNKT) cells, are characterized by their rapid activation in response to non-peptide antigens, such as lipids. While the transcriptional profiles of naive, effector and memory adaptive T cells have been well studied, less is known about transcriptional regulation of different iNKT cell activation states. Here, using single cell RNA-sequencing, we performed longitudinal profiling of activated murine iNKT cells, generating a transcriptomic atlas of iNKT cell activation states. We found that transcriptional signatures of activation are highly conserved among heterogeneous iNKT cell populations, including NKT1, NKT2 and NKT17 subsets, and human iNKT cells. Strikingly, we found that regulatory iNKT cells, such as adipose iNKT cells, undergo blunted activation, and display constitutive enrichment of memory-like cMAF+ and KLRG1+ populations. Moreover, we identify a conserved cMAF-associated transcriptional network among NKT10 cells, providing novel insights into the biology of regulatory and antigen experienced iNKT cells.