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Unpaired cysteine insertions favor transmembrane dimerization and induce ligand-independent constitutive cytokine receptor signaling

  • Lynn Affrica Felicitas Baumgärtner , Julia Ettich , Helene Balles , Dorothee Johanna Lapp ORCID logo , Sofie Mossner , Christin Bassenge , Meryem Ouzin ORCID logo , Helmut Hanenberg ORCID logo , Jürgen Scheller ORCID logo and Doreen Manuela Floss ORCID logo EMAIL logo
Published/Copyright: May 3, 2024
Biological Chemistry
From the journal Biological Chemistry Volume 405 Issue 7-8

Abstract

Naturally occurring gain-of-function (GOF) mutants have been identified in patients for a variety of cytokine receptors. Although this constitutive activation of cytokine receptors is strongly associated with malignant disorders, ligand-independent receptor activation is also a useful tool in synthetic biology e.g. to improve adoptive cellular therapies with genetically modified T-cells. Balanced Interleukin (IL-)7 signaling via a heterodimer of IL-7 receptor (IL-7Rα) and the common γ-chain (γc) controls T- and B-cell development and expansion, whereas uncontrolled IL-7 signaling can drive acute lymphoid leukemia (ALL) development. The ALL-driver mutation PPCL in the transmembrane domain of IL-7Rα is a mutational insertion of the four amino acids proline-proline-cysteine-leucine and leads to ligand-independent receptor dimerization and constitutive activation. We showed here in the cytokine-dependent pre-B-cell line Ba/F3 that the PPCL-insertion in a synthetic version of the IL-7Rα induced γc-independent STAT5 and ERK phosphorylation and also proliferation of the cells and that booster-stimulation by arteficial ligands additionally generated non-canonical STAT3 phosphorylation via the synthetic IL-7Rα-PPCL-receptors. Transfer of the IL-7Rα transmembrane domain with the PPCL insertion into natural and synthetic cytokine receptor chains of the IL-6, IL-12 and Interferon families also resulted in constitutive receptor signaling. In conclusion, our data suggested that the insertion of the mutated PPCL IL-7Rα transmembrane domain is an universal approach to generate ligand-independent, constitutively active cytokine receptors.

Keywords: gain-of-function receptors; signal transduction; interleukin; synthetic receptors; receptor homo-dimerization
Corresponding author: Doreen Manuela Floss, Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany, E-mail:
Lynn Affrica Felicitas Baumgärtner and Julia Ettich contributed equally to this work. Dorothee Johanna Lapp: Present address: Institute of Innate Immunity, Medical Faculty, University of Bonn, D-53127 Bonn, Germany and Department of Microbiology & Immunology, The University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria 3000, Australia; Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany. Sofie Mossner: Present address: MLM Medical Labs, D-41066 Mönchengladbach, Germany; Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany. Meryem Ouzin: Present address: Institute of Transplantation Diagnostics and Cell Therapeutics, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany; Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany.

Funding source: Deutsche Forschungsgemeinschaft

Award Identifier / Grant number: SCHE 907/5–1

Award Identifier / Grant number: SCHE 907/6-1

Funding source: Deutsche Krebshilfe e.V.

Award Identifier / Grant number: 70114844

Acknowledgments

We thank Yvonne Arlt for technical assistance.

  1. Research ethics: Not applicable.

  2. Author contributions: The authors have accepted responsibility for the entire content of this manuscript and approved its submission. L.A.F.B., D.J.L., S.M., C.B., M.O. and H.B. formal analysis, validation, investigation; D.M.F. and J.E. supervision; J.E. resources; L.A.F.B. visualization; J.S. and D.M.F. conceptualization; J.S. and H.H. funding acquisition; L.A.F.B., J.E., H.H., J.S. and D.M F. writing and editing; D.M.F. and J.E. data curation; D.M.F. and J.E. methodology; L.A.F.B., J.S. and D.M.F. writing-original draft; D.M.F. project administration.

  3. Competing interests: The authors state no conflict of interest.

  4. Research funding: J.S. was funded by grants from the Deutsche Forschungsgemeinschaft (SCHE 907/5-1 and SCHE 907/6-1). H.H. acknowledges a grant from the Deutsche Krebshilfe e.V. (70114844).

  5. Data availability: Not applicable.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/hsz-2023-0344).

Received: 2023-11-10
Accepted: 2024-03-25
Published Online: 2024-05-03
Published in Print: 2024-07-26

© 2024 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/hsz-2023-0344
Keywords for this article
gain-of-function receptors; signal transduction; interleukin; synthetic receptors; receptor homo-dimerization

Articles in the same Issue

  1. Frontmatter
  2. Highlight: New developments in immunoengineering
  3. Highlight: new developments in immunoengineering
  4. Better safe than sorry: dual targeting antibodies for cancer immunotherapy
  5. Bovine ultralong CDR-H3 derived knob paratopes elicit potent TNF-α neutralization and enable the generation of novel adalimumab-based antibody architectures with augmented features
  6. Development of an enabling platform biotechnology for the production of proteins
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  8. Advances in preclinical TCR characterization: leveraging cell avidity to identify functional TCRs
  9. Unpaired cysteine insertions favor transmembrane dimerization and induce ligand-independent constitutive cytokine receptor signaling
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