mRNA Encoding a Bispecific Single Domain Antibody Construct Protects against Influenza A Virus Infection in Mice

Lien Van Hoecke; Rein Verbeke; Dorien De Vlieger; Heleen Dewitte; Kenny Roose; Sharon Van Nevel; Olga Krysko; Claus Bachert; Bert Schepens; Ine Lentacker; Xavier Saelens

doi:10.1016/j.omtn.2020.04.015

mRNA Encoding a Bispecific Single Domain Antibody Construct Protects against Influenza A Virus Infection in Mice

Mol Ther Nucleic Acids. 2020 Jun 5:20:777-787. doi: 10.1016/j.omtn.2020.04.015. Epub 2020 May 1.

Authors

Affiliations

¹ VIB-UGent Center for Medical Biotechnology, VIB, 9000 Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, 9000 Ghent, Belgium.
² Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium; Cancer Research Institute Ghent, 9000 Ghent, Belgium.
³ VIB-UGent Center for Medical Biotechnology, VIB, 9000 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, 9000 Ghent, Belgium.
⁴ Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium; Cancer Research Institute Ghent, 9000 Ghent, Belgium; Laboratory for Molecular and Cellular Therapy, Vrije Universiteit Brussel, 1090 Jette, Belgium.
⁵ Upper Airways Research Laboratory, Department of Head and Skin, Ghent University, 9000 Ghent, Belgium.
⁶ VIB-UGent Center for Medical Biotechnology, VIB, 9000 Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, 9000 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, 9000 Ghent, Belgium.
⁷ VIB-UGent Center for Medical Biotechnology, VIB, 9000 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, 9000 Ghent, Belgium. Electronic address: xavier.saelens@vib-ugent.be.

Abstract

To date, mRNA-based biologics have mainly been developed for prophylactic and therapeutic vaccination to combat infectious diseases or cancer. In the past years, optimization of the characteristics of in vitro transcribed mRNA has led to significant reduction of the inflammatory responses. Thanks to this, mRNA therapeutics have entered the field of passive immunization. Here, we established an mRNA treatment that is based on mRNA that codes for a bispecific single-domain antibody construct that can selectively recruit innate immune cells to cells infected with influenza A virus. The constructs consist of a single-domain antibody that binds to the ectodomain of the conserved influenza A matrix protein 2, while the other single-domain antibody binds to the activating mouse Fcγ receptor IV. Formulating the mRNA into DOTAP (1,2-dioleoyl-3-trimethylammonium-propane)/cholesterol nanoparticles and delivering these intratracheally to mice allowed the production of the bispecific single-domain antibody in the lungs, and administration of these mRNA-particles prior to influenza A virus infection was associated with a significant reduction in viral titers and a reduced morbidity in mice. Overall, our data provide evidence that the local delivery of mRNA encoding a bispecific single-domain antibody format in the lungs could be a promising pulmonary antiviral prophylactic treatment.

Keywords: Influeanza A; bispecific singel domain antibody; mRNA; mRNA therapeutics; matrix protein 2; nanoparticles; passive immunization.