pocketz.doi.bio/2025-12-11_142749_pubmed_37403357

A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation - PubMed

URL: https://pubmed.ncbi.nlm.nih.gov/37403357/

Saved: 2025-12-11T22:27:49.691Z

Mol Ther . 2023 Aug 2;31(8):2360-2375. doi: 10.1016/j.ymthe.2023.06.017. Epub 2023 Jul 3. A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation Taishi Kimura 1, Joseph M Leal 2, Adrian Simpson 2, Nikole L Warner 2, Bryan J Berube 2, Jacob F Archer 2, Stephanie Park 2, Ryan Kurtz 2, Troy Hinkley 2, Katrina Nicholes 2, Shibbu Sharma 2, Malcolm S Duthie 2, Peter Berglund 2, Steven G Reed 2, Amit P Khandhar 2, Jesse H Erasmus 3 Affiliations expand PMID: 37403357 PMCID: PMC10422015 DOI: 10.1016/j.ymthe.2023.06.017 Full text linksCite Abstract

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.

Keywords: LION formulation; RNA vaccines; infectious diseases; innate immunity; lipid nanoparticles; multivalent vaccines; nanoparticle emulsion; reactogenicity; replicating RNA; side-effect.

PubMed Disclaimer

Conflict of interest statement

Declaration of interests T.K., J.M.L., A.S., N.L.W., S.P., R.K., J.F.A., B.J.B., T.H., K.N., S.S., A.K., M.S.D., P.B., S.G.R., and J.H.E. have equity interest in HDT Bio. J.H.E. is a consultant for InBios. P.B. is a consultant for Karkinox, and Chimeron. P.B. and J.H.E. are co-inventors on US patent application no. 63/011,860 “Nucleic acid vaccines against COVID-19” pertaining to repRNA-CoV-2S. J.H.E., A.K., and S.G.R. are co-inventors on US patent application no. 62/993,307 “Compositions and methods for delivery of RNA” pertaining to the LION formulation.

Figures

Graphical abstract

Figure 1

Intramuscular injection of repRNA/LION restricts…

Figure 2

Intramuscular injection of repRNA/LION activates…

Figure 3

Intramuscular injection of repRNA/LION induces…

Figure 4

Intramuscular injection of repRNA/LION induces…

Figure 5

Intramuscular injection of multivalent repRNA/LION…

Similar articles Antigen-dependent interplay of formulation, systemic innate responses, and antibody responses to multi-component replicon RNA vaccination. Kimura T, Reed SJ, Warner NL, Fredericks MN, Lewis TB, Lafferty A, Hodge E, Simpson A, Hinkley T, Khandhar AP, Fuller DH, Erasmus JH. Mol Ther Nucleic Acids. 2025 Jun 9;36(3):102595. doi: 10.1016/j.omtn.2025.102595. eCollection 2025 Sep 9. PMID: 40606648Free PMC article. Polyethylenimine-based polyplex delivery of self-replicating RNA vaccines. Démoulins T, Milona P, Englezou PC, Ebensen T, Schulze K, Suter R, Pichon C, Midoux P, Guzmán CA, Ruggli N, McCullough KC. Nanomedicine. 2016 Apr;12(3):711-722. doi: 10.1016/j.nano.2015.11.001. Epub 2015 Dec 1. PMID: 26592962 Self-Amplifying Pestivirus Replicon RNA Encoding Influenza Virus Nucleoprotein and Hemagglutinin Promote Humoral and Cellular Immune Responses in Pigs. Démoulins T, Ruggli N, Gerber M, Thomann-Harwood LJ, Ebensen T, Schulze K, Guzmán CA, McCullough KC. Front Immunol. 2021 Jan 28;11:622385. doi: 10.3389/fimmu.2020.622385. eCollection 2020. PMID: 33584723Free PMC article. Self-Replicating RNA Vaccine Delivery to Dendritic Cells. Démoulins T, Englezou PC, Milona P, Ruggli N, Tirelli N, Pichon C, Sapet C, Ebensen T, Guzmán CA, McCullough KC. Methods Mol Biol. 2017;1499:37-75. doi: 10.1007/978-1-4939-6481-9_3. PMID: 27987142Review. Replicon RNA vaccines: design, delivery, and immunogenicity in infectious diseases and cancer. Tang L, Que H, Wei Y, Yang T, Tong A, Wei X. J Hematol Oncol. 2025 Apr 17;18(1):43. doi: 10.1186/s13045-025-01694-2. PMID: 40247301Free PMC article.Review. See all similar articles Cited by Controlling reactogenicity while preserving immunogenicity from a self-amplifying RNA vaccine by modulating nucleocytoplasmic transport. Wojcechowskyj JA, Jong RM, Mäger I, Flach B, Munson PV, Mukherjee PP, Mertins B, Barcay KR, Folliard T. NPJ Vaccines. 2025 Apr 29;10(1):85. doi: 10.1038/s41541-025-01135-8. PMID: 40301369Free PMC article. Dissolving microneedles for nucleic acid delivery: A systematic search, review, and data synthesis. Tobos CI, Woodrow KA. Acta Biomater. 2025 Jun 15;200:115-131. doi: 10.1016/j.actbio.2025.05.025. Epub 2025 May 9. PMID: 40349901 A replicating RNA vaccine protects against severe fever with thrombocytopenia syndrome virus infection in mice. Stamper A, Bisom T, Meade-White K, Lewis M, Leventhal S, Clancy C, Hinkley T, Erasmus J, Feldmann H, Hawman DW. NPJ Vaccines. 2025 Oct 16;10(1):219. doi: 10.1038/s41541-025-01269-9. PMID: 41102248Free PMC article. mRNA vaccine designs for optimal adjuvanticity and delivery. Mochida Y, Uchida S. RNA Biol. 2024 Jan;21(1):1-27. doi: 10.1080/15476286.2024.2333123. Epub 2024 Mar 26. PMID: 38528828Free PMC article.Review. An anti-Shiga toxin VHH nanobody multimer protects mice against fatal toxicosis when administered intramuscularly as repRNA. Robinson SR, Dayao DA, Medina JA, Martone CJ, Yauch AK, Hinkley T, Erasmus JH, Shoemaker CB, Tzipori S. Infect Immun. 2024 Nov 12;92(11):e0023924. doi: 10.1128/iai.00239-24. Epub 2024 Oct 11. PMID: 39392311Free PMC article. See all "Cited by" articles References Pardi N., Hogan M.J., Porter F.W., Weissman D. mRNA vaccines - a new era in vaccinology. Nat. Rev. Drug Discov. 2018;17:261–279. doi: 10.1038/nrd.2017.243. - DOI - PMC - PubMed Hou X., Zaks T., Langer R., Dong Y. Lipid nanoparticles for mRNA delivery. Nat. Rev. Mater. 2021;6:1078–1094. doi: 10.1038/s41578-021-00358-0. - DOI - PMC - PubMed Bloom K., van den Berg F., Arbuthnot P. Self-amplifying RNA vaccines for infectious diseases. Gene Ther. 2021;28:117–129. doi: 10.1038/s41434-020-00204-y. - DOI - PMC - PubMed Vogel A.B., Lambert L., Kinnear E., Busse D., Erbar S., Reuter K.C., Wicke L., Perkovic M., Beissert T., Haas H., et al. Self-Amplifying RNA Vaccines Give Equivalent Protection against Influenza to mRNA Vaccines but at Much Lower Doses. Mol. Ther. 2018;26:446–455. doi: 10.1016/j.ymthe.2017.11.017. - DOI - PMC - PubMed Jackson L.A., Anderson E.J., Rouphael N.G., Roberts P.C., Makhene M., Coler R.N., McCullough M.P., Chappell J.D., Denison M.R., Stevens L.J., et al. An mRNA Vaccine against SARS-CoV-2 - Preliminary Report. N. Engl. J. Med. 2020;383:1920–1931. doi: 10.1056/NEJMoa2022483. - DOI - PMC - PubMed Show all 57 references Publication types Research Support, N.I.H., Extramural MeSH terms Animals Antigens Humans Immunity, Humoral Inflammation Mice Nanoparticles* RNA* / genetics Tissue Distribution Substances RNA Antigens Related information Gene MedGen Protein (RefSeq) Grants and funding R61 AI161811/AI/NIAID NIH HHS/United States 75N93019C00037/AI/NIAID NIH HHS/United States 75N93019C00008/AI/NIAID NIH HHS/United States R43 AI165100/AI/NIAID NIH HHS/United States 75N93020C00028/AI/NIAID NIH HHS/United States LinkOut - more resources Full Text Sources Elsevier Science PubMed Central Other Literature Sources The Lens - Patent Citations Database Molecular Biology Databases Mouse Genome Informatics (MGI)