The proposal is to develop an RNA-based multi-antigen SARS-CoV-2 vaccine, supporting NIAID's call to developa vaccine using emerging antigen design strategies and novel delivery approaches. The SARS-CoV-2 pandemicis actively threatening public health, world economies, and ways of life. Vaccine candidates are in human trialsnow, but long-term solutions will require an adaptable vaccine system that can be universalized broadly to allmembers of the coronavirus family. Tiba's vaccine will be innovative in two respects: 1) The inclusion of all fourstructural virion proteins will allow a deeper and potentially broader immune response against coronavirus, asmultiple conserved T cell epitopes will be presented by the vaccine. In addition, the incorporation of structuralproteins besides the Spike (S) protein will allow the formation of immunogenic VLPs in situ. 2) Conventional lipidnanoparticles (LNPs), which are the mainstay of nucleic acid delivery, require a large proportion of "structural"lipid, resulting in a relatively low RNA content. Tiba has developed a modified dendrimer delivery molecule thatmaximizes RNA mass content, protects RNA from degradation, and enables efficient uptake by cells. Thisapproach will increase immunogenicity and thus effectiveness of the vaccine. The first Phase 1 Aim is to optimizecandidate payloads comprising M, N, and E mRNA or saRNA. Conventional mRNAs encoding the M, N, and Eproteins will be codon-optimized and lead RNA payloads will then be formulated with Tiba's modified dendrimerdelivery material. With the M, N, and E payloads generated in both saRNA and mRNA formats, effectiveimmunogenic doses for each will be determined by injecting six-week-old BALB/c mice with nanoparticles atdifferent doses. The antigen RNA payloads to proceed for titration into S RNA vaccine candidates will beselected. The minimum effective dose for each M, N, and E mRNA or saRNA will be roughly estimated using aregression model. Aim 2 will test the lead candidate payloads along with S in mice. Combination formulationswill be generated wherein different masses of the lead M, N, and E RNAs are titrated into the optimal S mRNAor saRNA formula to identify a formulation mixture that maximizes M, N, and E cellular immune responses whilenot suppressing the S humoral response. Treatment doses will deliver the same mass of S RNA across allgroups, and that optimal mass will be chosen based on the results of dose-finding preliminary studies. Using theresults of the study, two mixtures that confer detectable responses against each virion component while notsuppressing S antibody and T cell responses will be selected as lead candidates. For Aim 3, the lead candidateswill be used in a hamster challenge study. Vaccinations will be performed by i.m. injection in golden Syrianhamsters. Challenges will be performed using the current best practice. The benchmark of success for this studyis evidence of immune responses against firstly the S component of the vaccine, and concomitant protectionagainst disease, ideally after a single administration. If successful, further development of lead vaccinecandidates and the path to commercialization will be forthcoming in a Phase 2 SBIR Proposal. Project Narrative Development of an effective vaccine against coronavirus is critical for world economies, devastated by the current pandemic, to begin recovery and to save millions from significant illness or death. We propose to produce a multi-antigen RNA-based prototype vaccine against SARS-CoV-2, utilizing a novel and innovative modified- dendrimer-based nanoparticle delivery technology that will solve the challenges hindering current single-antigen nucleic acid vaccine approaches and potentially lead to more broad protection against coronaviruses. The proposed vaccine product can be easily adapted to include sequences of other coronaviruses with pandemic potential due to its modular nature, making it a potential approach to multivalent or universal coronavirus vaccination. 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