Kinetics of cytokine receptor trafficking determine signaling and functional selectivity

  1. Jonathan Martinez-Fabregas
  2. Stephan Wilmes
  3. Luopin Wang
  4. Maximillian Hafer
  5. Elizabeth Pohler
  6. Juliane Lokau
  7. Christoph Garbers
  8. Adeline Cozzani
  9. Jacob Piehler
  10. Majid Kazemian  Is a corresponding author
  11. Suman Mitra  Is a corresponding author
  12. Ignacio Moraga Gonzalez  Is a corresponding author
  1. University of Dundee, United Kingdom
  2. Purdue University, United States
  3. University of Osnabrück, Germany
  4. Otto-von-Guericke-University Magdeburg, Germany
  5. Université de Lille, France

Abstract

Cytokines activate signaling via assembly of cell surface receptors, but it is unclear whether modulation of cytokine-receptor binding parameters can modify biological outcomes. We have engineered IL-6 variants with different affinities to gp130 to investigate how cytokine receptor binding dwell-times influence functional selectivity. Engineered IL-6 variants showed a range of signaling amplitudes and induced biased signaling, with changes in receptor binding dwell-times affecting more profoundly STAT1 than STAT3 phosphorylation. We show that this differential signaling arises from defective translocation of ligand-gp130 complexes to the endosomal compartment and competitive STAT1/STAT3 binding to phospho-tyrosines in gp130, and results in unique patterns of STAT3 binding to chromatin. This leads to a graded gene expression response and differences in ex vivo differentiation of Th17, Th1 and Treg cells. These results provide a molecular understanding of signaling biased by cytokine receptors, and demonstrate that manipulation of signaling thresholds is a useful strategy to decouple cytokine functional pleiotropy.

Data availability

Sequencing data have been deposited in GEO under accession number code: GSE130810

The following data sets were generated

Article and author information

Author details

  1. Jonathan Martinez-Fabregas

    Division of Cell Signaling and Immunology, School of Life Sciences, University of Dundee, Dundee, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5809-065X
  2. Stephan Wilmes

    Division of Cell Signaling and Immunology, School of Life Sciences, University of Dundee, Dundee, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Luopin Wang

    Department of Biochemistry, Purdue University, West Lafayette, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Maximillian Hafer

    Department of Biology, University of Osnabrück, Osnabrück, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0853-2637
  5. Elizabeth Pohler

    Division of Cell Signaling and Immunology, School of Life Sciences, University of Dundee, Dundee, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  6. Juliane Lokau

    Department of Pathology, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2573-7282
  7. Christoph Garbers

    Department of Pathology, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Adeline Cozzani

    Université de Lille, Lille, France
    Competing interests
    The authors declare that no competing interests exist.
  9. Jacob Piehler

    Department of Biology, University of Osnabrück, Osnabrück, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Majid Kazemian

    Department of Biochemistry, Purdue University, West Lafayette, United States
    For correspondence
    kazemian@purdue.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7080-8820
  11. Suman Mitra

    Université de Lille, Lille, France
    For correspondence
    suman.mitra@inserm.fr
    Competing interests
    The authors declare that no competing interests exist.
  12. Ignacio Moraga Gonzalez

    Division of Cell Signaling and Immunology, School of Life Sciences, University of Dundee, Dundee, United Kingdom
    For correspondence
    IMoragagonzalez@dundee.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9909-0701

Funding

Horizon 2020 Framework Programme (714680)

  • Ignacio Moraga Gonzalez

Horizon 2020 Framework Programme (714680)

  • Jonathan Martinez-Fabregas

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

Reviewing Editor

  1. Philippe IH Bastiaens, Max Planck Institute of Molecular Physiology, Germany

Publication history

  1. Received: June 13, 2019
  2. Accepted: November 24, 2019
  3. Accepted Manuscript published: November 27, 2019 (version 1)
  4. Version of Record published: December 16, 2019 (version 2)

Copyright

© 2019, Martinez-Fabregas 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. Jonathan Martinez-Fabregas
  2. Stephan Wilmes
  3. Luopin Wang
  4. Maximillian Hafer
  5. Elizabeth Pohler
  6. Juliane Lokau
  7. Christoph Garbers
  8. Adeline Cozzani
  9. Jacob Piehler
  10. Majid Kazemian
  11. Suman Mitra
  12. Ignacio Moraga Gonzalez
(2019)
Kinetics of cytokine receptor trafficking determine signaling and functional selectivity
eLife 8:e49314.
https://doi.org/10.7554/eLife.49314

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