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Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors

Cytokines are small secreted proteins that among many functions also play key roles in the orchestration of inflammation in host defense and disease. Over the past years, a large number of biologics have been developed to target cytokines in disease, amongst which soluble receptor fusion proteins ha...

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Autores principales: Holgado, Aurora, Braun, Harald, Verstraete, Kenneth, Vanneste, Domien, Callewaert, Nico, Savvides, Savvas N., Afonina, Inna S., Beyaert, Rudi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7370943/
https://www.ncbi.nlm.nih.gov/pubmed/32754154
http://dx.doi.org/10.3389/fimmu.2020.01422
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author Holgado, Aurora
Braun, Harald
Verstraete, Kenneth
Vanneste, Domien
Callewaert, Nico
Savvides, Savvas N.
Afonina, Inna S.
Beyaert, Rudi
author_facet Holgado, Aurora
Braun, Harald
Verstraete, Kenneth
Vanneste, Domien
Callewaert, Nico
Savvides, Savvas N.
Afonina, Inna S.
Beyaert, Rudi
author_sort Holgado, Aurora
collection PubMed
description Cytokines are small secreted proteins that among many functions also play key roles in the orchestration of inflammation in host defense and disease. Over the past years, a large number of biologics have been developed to target cytokines in disease, amongst which soluble receptor fusion proteins have shown some promise in pre-clinical studies. We have previously shown proof-of-concept for the therapeutic targeting of interleukin (IL)-33 in airway inflammation using a newly developed biologic, termed IL-33trap, comprising the ectodomains of the cognate receptor ST2 and the co-receptor IL-1RAcP fused into a single-chain recombinant fusion protein. Here we extend the biophysical and biological characterization of IL-33trap variants, and show that IL-33trap is a stable protein with a monomeric profile both at physiological temperatures and during liquid storage at 4°C. Reducing the N-glycan heterogeneity and complexity of IL-33trap via GlycoDelete engineering neither affects its stability nor its inhibitory activity against IL-33. We also report that IL-33trap specifically targets biologically active IL-33 splice variants. Finally, we document the generation and antagonistic activity of a single-chain IL-4/13trap, which inhibits both IL-4 and IL-13 signaling. Collectively, these results illustrate that single-chain soluble receptor fusion proteins against IL-4, IL-13, and IL-33 are novel biologics that might not only be of interest for research purposes and further interrogation of the role of their target cytokines in physiology and disease, but may also complement monoclonal antibodies for the treatment of allergic and other inflammatory diseases.
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spelling pubmed-73709432020-08-03 Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors Holgado, Aurora Braun, Harald Verstraete, Kenneth Vanneste, Domien Callewaert, Nico Savvides, Savvas N. Afonina, Inna S. Beyaert, Rudi Front Immunol Immunology Cytokines are small secreted proteins that among many functions also play key roles in the orchestration of inflammation in host defense and disease. Over the past years, a large number of biologics have been developed to target cytokines in disease, amongst which soluble receptor fusion proteins have shown some promise in pre-clinical studies. We have previously shown proof-of-concept for the therapeutic targeting of interleukin (IL)-33 in airway inflammation using a newly developed biologic, termed IL-33trap, comprising the ectodomains of the cognate receptor ST2 and the co-receptor IL-1RAcP fused into a single-chain recombinant fusion protein. Here we extend the biophysical and biological characterization of IL-33trap variants, and show that IL-33trap is a stable protein with a monomeric profile both at physiological temperatures and during liquid storage at 4°C. Reducing the N-glycan heterogeneity and complexity of IL-33trap via GlycoDelete engineering neither affects its stability nor its inhibitory activity against IL-33. We also report that IL-33trap specifically targets biologically active IL-33 splice variants. Finally, we document the generation and antagonistic activity of a single-chain IL-4/13trap, which inhibits both IL-4 and IL-13 signaling. Collectively, these results illustrate that single-chain soluble receptor fusion proteins against IL-4, IL-13, and IL-33 are novel biologics that might not only be of interest for research purposes and further interrogation of the role of their target cytokines in physiology and disease, but may also complement monoclonal antibodies for the treatment of allergic and other inflammatory diseases. Frontiers Media S.A. 2020-07-13 /pmc/articles/PMC7370943/ /pubmed/32754154 http://dx.doi.org/10.3389/fimmu.2020.01422 Text en Copyright © 2020 Holgado, Braun, Verstraete, Vanneste, Callewaert, Savvides, Afonina and Beyaert. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Holgado, Aurora
Braun, Harald
Verstraete, Kenneth
Vanneste, Domien
Callewaert, Nico
Savvides, Savvas N.
Afonina, Inna S.
Beyaert, Rudi
Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title_full Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title_fullStr Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title_full_unstemmed Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title_short Single-Chain Soluble Receptor Fusion Proteins as Versatile Cytokine Inhibitors
title_sort single-chain soluble receptor fusion proteins as versatile cytokine inhibitors
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7370943/
https://www.ncbi.nlm.nih.gov/pubmed/32754154
http://dx.doi.org/10.3389/fimmu.2020.01422
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