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bioRxiv · 10.1101/2025.06.27.661928

Nodamuravirus protein B2 boosts self-amplifying mRNA efficacy by overcoming innate immune barriers.

Sanchez-David, R. Y.·Le, H. D.·Nemegeer, J.·Goncalves, A.·Merits, A.·Maillard, P. V.
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Abstract

Self-amplifying RNA (sa-RNA) technology is a promising strategy for vaccine design, as its intracellular replication boosts transgene expression and provides self-adjuvanticity. However, sa-RNA efficiency is limited by innate immune responses triggered by the presence of intracellular double-stranded RNA (dsRNA). In vertebrates, differentiated cells mainly use type I interferon (IFN) system for protection against viruses, while stem cells rely on IFN-independent mechanisms such as antiviral RNA interference (RNAi). Here, we found that the efficiency of sa-RNAs based on chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) genomes is enhanced when co-expressed in cis with the Nodamura virus (NoV) B2 protein, a viral suppressor of RNAi. In stem cells, NoV B2 prevents Dicer-mediated processing of dsRNA, while in somatic cells, it blocks the translation shutdown caused by protein kinase R (PKR), a key effector of the IFN system. Notably, NoV B2 does not interfere with IFN induction and signalling, preserving sa-RNAs self-adjuvant properties. Mechanistically, NoV B2 sequesters replication-derived dsRNA at the cell periphery, offering a novel strategy to boost sa-RNA efficiency without compromising its immune stimulatory properties. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=130 SRC="FIGDIR/small/661928v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@4412aforg.highwire.dtl.DTLVardef@126d089org.highwire.dtl.DTLVardef@144dd38org.highwire.dtl.DTLVardef@179bac_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Sanchez-David, R. Y., Le, H. D., Nemegeer, J., Goncalves, A., Merits, A., Maillard, P. V.. 2025-07-02. Nodamuravirus protein B2 boosts self-amplifying mRNA efficacy by overcoming innate immune barriers.. https://doi.org/10.1101/2025.06.27.661928

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