Innate inhibiting proteins enhance expression and immunogenicity of self-amplifying RNA
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Published version
Author(s)
Type
Journal Article
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. Here, we screen a library of saRNA constructs with cis-encoded innate inhibiting proteins (IIPs) and determine the effect on protein expression and immunogenicity. We observed that the PIV-5 V and MERS-CoV ORF4a proteins enhance protein expression 100-500-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 JAK/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 RABV-specific IgG titer and neutralization IC50 by ~10-fold in rabbits, but not 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.
Date Issued
2021-03-03
Date Acceptance
2020-11-05
Citation
Molecular Therapy, 2021, 29 (3), pp.1174-1185
ISSN
1525-0016
Publisher
Cell Press
Start Page
1174
End Page
1185
Journal / Book Title
Molecular Therapy
Volume
29
Issue
3
Copyright Statement
© 2020 This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Commission of the European Communities
Identifier
https://www.sciencedirect.com/science/article/pii/S1525001620306092?via%3Dihub
Grant Number
EP/R013764/1
794059
Subjects
RNA
gene delivery
immunomodulation
innate immunity
interferon
nanoparticles
replicon
self-amplifying
vaccines
06 Biological Sciences
10 Technology
11 Medical and Health Sciences
Biotechnology
Publication Status
Published
Date Publish Online
2020-12-21