A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation
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RNA vaccines possess significant clinical promise in counteracting human diseases caused by infectious or cancerous threats. Self-amplifying replicon RNA (repRNA) has been thought to offer the potential for enhanced potency and dose sparing. However, repRNA is a potent trigger of innate immune responses in vivo, which can cause reduced transgene expression and dose-limiting reactogenicity, as highlighted by recent clinical trials. Here, we report that multivalent repRNA vaccination, necessitating higher doses of total RNA, could be safely achieved in mice by delivering multiple repRNAs with a localizing cationic nanocarrier formulation (LION). Intramuscular delivery of multivalent repRNA by LION resulted in localized biodistribution accompanied by significantly upregulated local innate immune responses and the induction of antigen-specific adaptive immune responses in the absence of systemic inflammatory responses. In contrast, repRNA delivered by lipid nanoparticles (LNPs) showed generalized biodistribution, a systemic inflammatory state, an increased body weight loss, and failed to induce neutralizing antibody responses in a multivalent composition. These findings suggest that in vivo delivery of repRNA by LION is a platform technology for safe and effective multivalent vaccination through mechanisms distinct from LNP-formulated repRNA vaccines.
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Previous efforts to decrease immunogenicity of saRNA focused on sequence evolution, co-expression of viral inhibitory proteins and optimization of the delivery vehicle.
“Previous efforts to decrease the immunogenicity of saRNA focused on sequence evolution, co-expression of viral inhibitory proteins and optimization of the delivery vehicle16–18.”
From Complete substitution with modified nucleotides in self-amplifying RNA suppresses the interferon response and increases potency (2025) · cites this paper as [18]
The Discussion (page unavailable) supports delivery-vehicle optimization as an alternative way to limit systemic innate responses to replicating RNA. The Results discuss prior efforts to inhibit type I interferon signaling, and the References name work on innate-inhibiting proteins, but our copy does not describe co-expression of viral inhibitory proteins. The Discussion says sequence optimization cannot be applied to replicating RNA to raise its dose ceiling, rather than identifying it as a prior saRNA strategy. (LLM, conf=0.85)
Experimental — accuracy not yet measuredThe repRNA/LIONTM platform is less reactogenic than traditional LNPs.
“The repRNA/LION™ platform is less reactogenic than traditional LNPs suggesting higher tolerable single vaccine doses would be possible33.”
From Clade 2.3.4.4b but not historical clade 1 HA replicating RNA vaccine protects against bovine H5N1 challenge in mice (2025) · cites this paper as [33]
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Self-citation
- Authors
- Kimura, T., Leal, J. M., Simpson, A., Warner, N. L., Berube, B. J., Archer, J. F., Park, S., Kurtz, R., Hinkley, T., Nicholes, K., Sharma, S., Duthie, M. S., Berglund, P., Reed, S. G., Khandhar, A. P., & Erasmus, J. H.
- Affiliations
- University of Washington (US)
- Corresponding author
- Kimura, Taishi
- Erasmus, Jesse H.
- ORCID
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2023-08
- Volume
- 31
- Issue
- 8
- Pages
- 2360-2375
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC10422015
- OpenAlex
- W4382933450
- Cited by
- 52
- References
- 58
- Citations by year
- 2026: 8 · 2025: 21 · 2024: 18 · 2023: 5
- FWCI
- 4.80
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (75N93019C00008, R61AI161811, 75N93019C00037, 1R43AI165100-01, 75N93020C00028, 62/993,307)
- 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
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Keywords
- RNA vaccines, innate immune response, systemic inflammation, lipid nanoparticles, intramuscular administration, neutralizing antibodies, vaccine reactogenicity
- MeSH terms
- Animals, Antigens, Humans, Inflammation, RNA, Tissue Distribution, Mice, Nanoparticles, Immunity, Humoral
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