Cited by 105RNA Interference and Gene DeliveryImmunotherapy and Immune Responsesinterferon and immune responses

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Innate Inhibiting Proteins Enhance Expression and Immunogenicity of Self-Amplifying RNA

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Blakney, Anna K., McKay, Paul F., Bouton, Clément R., Hu, Kai, Samnuan, Karnyart, Shattock, Robin J.·2021·Molecular Therapy

DOI 10.1016/j.ymthe.2020.11.011PMC PMC7935664

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Abstract

Self-amplifying RNA (saRNA) is a cutting-edge platform for both nucleic acid vaccines and therapeutics. saRNA is self-adjuvanting, as it activates types I and III interferon (IFN), which enhances the immunogenicity of RNA vaccines but can also lead to inhibition of translation. In this study, we screened a library of saRNA constructs with cis -encoded innate inhibiting proteins (IIPs) and determined the effect on protein expression and immunogenicity. We observed that the PIV-5 V and Middle East respiratory syndrome coronavirus (MERS-CoV) ORF4a proteins enhance protein expression 100- to 500-fold in vitro in IFN-competent HeLa and MRC5 cells. We found that the MERS-CoV ORF4a protein partially abates dose nonlinearity in vivo , and that ruxolitinib, a potent Janus kinase (JAK)/signal transducer and activator of transcription (STAT) inhibitor, but not the IIPs, enhances protein expression of saRNA in vivo . Both the PIV-5 V and MERS-CoV ORF4a proteins were found to enhance the percentage of resident cells in human skin explants expressing saRNA and completely rescued dose nonlinearity of saRNA. Finally, we observed that the MERS-CoV ORF4a increased the rabies virus ( RABV )-specific immunoglobulin G (IgG) titer and neutralization half-maximal inhibitory concentration (IC 50 ) by ∼10-fold in rabbits, but not in mice or rats. These experiments provide a proof of concept that IIPs can be directly encoded into saRNA vectors and effectively abate the nonlinear dose dependency and enhance immunogenicity.

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1 retracted

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Authors: Blakney, A. K., McKay, P. F., Bouton, C. R., Hu, K., Samnuan, K., & Shattock, R. J.
Affiliations: Imperial College London (GB)
Corresponding author: Blakney, Anna K.
Shattock, Robin J.
ORCID: Blakney, Anna K.
McKay, Paul F.
Bouton, Clément R.
Hu, Kai
Samnuan, Karnyart
Shattock, Robin J.
Venue: Molecular Therapy
Publisher: Elsevier BV
Published: 2021-03
Volume: 29
Issue: 3
Pages: 1174-1185
ISSN: 1525-0016 (ISSN-L)
DOI: 10.1016/j.ymthe.2020.11.011
PMC: PMC7935664
OpenAlex: W3115814127
Cited by: 105
References: 60
Citations by year: 2026: 23 · 2025: 22 · 2024: 23 · 2023: 15 · 2022: 16 · 2021: 6
FWCI: 3.00
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor)
Locations: Molecular Therapy (publishedVersion, cc-by-nc-nd)
PubMed Central (publishedVersion, cc-by-nc-nd)
PubMed (publishedVersion)
Spiral (Imperial College London) (submittedVersion, cc-by-nc-nd)
Funders: H2020 Marie Skłodowska-Curie Actions
Engineering and Physical Sciences Research Council
Whitaker International Fellows and Scholars Program
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
Keywords: self-amplifying RNA, rabies virus, interferon, protein synthesis inhibition, ruxolitinib, JAK/STAT inhibition, rabies virus neutralizing antibodies
MeSH terms: Immunogenicity, Vaccine, mRNA Vaccines, Animals, Cell Line, Encephalitis Virus, Venezuelan Equine, Fibroblasts, Gene Expression Regulation, HeLa Cells, Humans, Immunoglobulin G, Immunity, Innate, Nitriles, Pyrazoles, Pyrimidines, Rabbits, Rabies virus, Protein Biosynthesis, Vaccines, Synthetic, Viral Envelope Proteins, Signal Transduction, NF-kappa B, Parainfluenza Virus 5, STAT Transcription Factors, Interferon Regulatory Factor-3, Mice, Rats, Janus Kinases, Host-Pathogen Interactions, Middle East Respiratory Syndrome Coronavirus

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Innate Inhibiting Proteins Enhance Expression and Immunogenicity of Self-Amplifying RNA

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