Engineered extracellular vesicle decoy receptor-mediated modulation of the IL6 trans-signalling pathway in muscle

Mariana Conceição; Laura Forcina; Oscar P B Wiklander; Dhanu Gupta; Joel Z Nordin; Besarte Vrellaku; Graham McClorey; Imre Mäger; André Gӧrgens; Per Lundin; Antonio Musarò; Matthew J A Wood; Samir El Andaloussi; Thomas C Roberts

doi:10.1016/j.biomaterials.2020.120435

Engineered extracellular vesicle decoy receptor-mediated modulation of the IL6 trans-signalling pathway in muscle

Biomaterials. 2021 Jan:266:120435. doi: 10.1016/j.biomaterials.2020.120435. Epub 2020 Oct 5.

Authors

Affiliations

¹ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Department of Paediatrics, University of Oxford, Oxford, UK. Electronic address: mariana.conceicao@paediatrics.ox.ac.uk.
² DAHFMO-Unit of Histology and Medical Embryology, Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci Bolognetti, IMM, Sapienza University of Rome, Rome, Italy.
³ Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Evox Therapeutics Limited, Oxford Science Park, Oxford, UK.
⁴ Department of Paediatrics, University of Oxford, Oxford, UK.
⁵ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Department of Paediatrics, University of Oxford, Oxford, UK.
⁶ Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Evox Therapeutics Limited, Oxford Science Park, Oxford, UK; Institute for Transfusion Medicine, University Hospital Essen, Essen, Germany.
⁷ Evox Therapeutics Limited, Oxford Science Park, Oxford, UK.
⁸ DAHFMO-Unit of Histology and Medical Embryology, Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci Bolognetti, IMM, Sapienza University of Rome, Rome, Italy; Center for Life Nano Science @Sapienza, Istituto Italiano di Tecnologia, Rome, Italy.
⁹ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Department of Paediatrics, University of Oxford, Oxford, UK; MDUK Oxford Neuromuscular Centre, University of Oxford, Oxford, UK.
¹⁰ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Evox Therapeutics Limited, Oxford Science Park, Oxford, UK.
¹¹ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Department of Paediatrics, University of Oxford, Oxford, UK; MDUK Oxford Neuromuscular Centre, University of Oxford, Oxford, UK. Electronic address: thomas.roberts@paediatrics.ox.ac.uk.

PMID: 33049461
DOI: 10.1016/j.biomaterials.2020.120435

Abstract

The cytokine interleukin 6 (IL6) is a key mediator of inflammation that contributes to skeletal muscle pathophysiology. IL6 activates target cells by two main mechanisms, the classical and trans-signalling pathways. While classical signalling is associated with the anti-inflammatory activities of the cytokine, the IL6 trans-signalling pathway mediates chronic inflammation and is therefore a target for therapeutic intervention. Extracellular vesicles (EVs) are natural, lipid-bound nanoparticles, with potential as targeted delivery vehicles for therapeutic macromolecules. Here, we engineered EVs to express IL6 signal transducer (IL6ST) decoy receptors to selectively inhibit the IL6 trans-signalling pathway. The potency of the IL6ST decoy receptor EVs was optimized by inclusion of a GCN4 dimerization domain and a peptide sequence derived from syntenin-1 which targets the decoy receptor to EVs. The resulting engineered EVs were able to efficiently inhibit activation of the IL6 trans-signalling pathway in reporter cells, while having no effect on the IL6 classical signalling. IL6ST decoy receptor EVs, were also capable of blocking the IL6 trans-signalling pathway in C2C12 myoblasts and myotubes, thereby inhibiting the phosphorylation of STAT3 and partially reversing the anti-differentiation effects observed when treating cells with IL6/IL6R complexes. Treatment of a Duchenne muscular dystrophy mouse model with IL6ST decoy receptor EVs resulted in a reduction in STAT3 phosphorylation in the quadriceps and gastrocnemius muscles of these mice, thereby demonstrating in vivo activity of the decoy receptor EVs as a potential therapy. Taken together, this study reveals the IL6 trans-signalling pathway as a promising therapeutic target in DMD, and demonstrates the therapeutic potential of IL6ST decoy receptor EVs.

Keywords: Decoy receptors; Exosomes; Extracellular vesicles; IL6; Inflammation; Muscle.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Extracellular Vesicles*
Interleukin-6
Mice
Muscle Fibers, Skeletal
Muscular Dystrophy, Duchenne*
Signal Transduction

Substances

Interleukin-6