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.

Metrics

  • 3,899
    Page views
  • 612
    Downloads
  • 22
    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. 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

Further reading

    1. Immunology and Inflammation
    Xing Feng, Ruifeng Sun ... Joao Pedro Pereira
    Research Article Updated

    Acute lymphoblastic and myeloblastic leukemias (ALL and AML) have been known to modify the bone marrow microenvironment and disrupt non-malignant hematopoiesis. However, the molecular mechanisms driving these alterations remain poorly defined. Using mouse models of ALL and AML, here we show that leukemic cells turn off lymphopoiesis and erythropoiesis shortly after colonizing the bone marrow. ALL and AML cells express lymphotoxin α1β2 and activate lymphotoxin beta receptor (LTβR) signaling in mesenchymal stem cells (MSCs), which turns off IL7 production and prevents non-malignant lymphopoiesis. We show that the DNA damage response pathway and CXCR4 signaling promote lymphotoxin α1β2 expression in leukemic cells. Genetic or pharmacological disruption of LTβR signaling in MSCs restores lymphopoiesis but not erythropoiesis, reduces leukemic cell growth, and significantly extends the survival of transplant recipients. Similarly, CXCR4 blocking also prevents leukemia-induced IL7 downregulation and inhibits leukemia growth. These studies demonstrate that acute leukemias exploit physiological mechanisms governing hematopoietic output as a strategy for gaining competitive advantage.

    1. Immunology and Inflammation
    Eve Richardson, Špela Binter ... Simon J Watson
    Research Article

    Immunoglobulin loci-transgenic animals are widely used in antibody discovery and increasingly in vaccine response modelling. In this study, we phenotypically characterised B-cell populations from the Intelliselect® Transgenic mouse (Kymouse) demonstrating full B-cell development competence. Comparison of the naïve B-cell receptor (BCR) repertoires of Kymice BCRs, naïve human, and murine BCR repertoires revealed key differences in germline gene usage and junctional diversification. These differences result in Kymice having CDRH3 length and diversity intermediate between mice and humans. To compare the structural space explored by CDRH3s in each species’ repertoire, we used computational structure prediction to show that Kymouse naïve BCR repertoires are more human-like than mouse-like in their predicted distribution of CDRH3 shape. Our combined sequence and structural analysis indicates that the naïve Kymouse BCR repertoire is diverse with key similarities to human repertoires, while immunophenotyping confirms that selected naïve B-cells are able to go through complete development.