Design, Testing, and Optimization of a Preclinical mRNA-LNP Vaccine for Hepatitis C Virus Prophylaxis
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Graduate group
Discipline
Immunology and Infectious Disease
Biology
Subject
Infectious Disease
mRNA
mRNA-LNP
Vaccines
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Abstract
Hepatitis C virus (HCV) presents an increasing threat to global public health. A vaccine able to protect against chronic HCV infection would prevent serious sequelae of infection and limit costs associated with long-term infection and treatment. Advances in techniques to study immune responses against HCV have uncovered the significant protective role of antibodies which can potently neutralize diverse viral strains. Concurrently, progress in HIV research has revealed mechanisms by which broadly neutralizing antibodies (bNAbs) develop in natural infection and vaccination. To evaluate these strategies for HCV, we paired these developments with advances in the nucleoside-modified mRNA lipid nanoparticle (mRNA-LNP) vaccine platform. We evaluated structural and nonstructural viral proteins in our vaccine constructs, demonstrating that encoding p7 along with core (C), envelope glycoprotein 1 (E1), and envelope glycoprotein 2 (E2) led to significantly more viral protein expression and secretion, as well as the secretion of HCV virus-like particles. Mouse studies showed that including p7 led to significantly higher binding titers against a panel of diverse heterologous HCV variants, as well as neutralization against heterologous strains. Employing a lineage design approach, we selected sequences from the viral quasispecies of an HCV+ individual who developed bNAbs which cleared their infection. We encoded the C, E1, E2, and p7 proteins and delivered them to mice according to various immunization strategies. The approach which combined immunogens derived from early-, mid-, and late-infection for each immunization elicited significantly higher binding and neutralizing antibody responses against a panel representing global HCV antigenic diversity. These findings indicate that combining immunogens selected to bind to the germline and intermediate B cells of a bNAb lineage yields significantly broader and more potent humoral responses than other approaches. The sum of these findings advances the development of a prophylactic HCV vaccine, deepens our understanding of bNAb elicitation through vaccination, and provides a proof of concept for the suitability of the mRNA-LNP platform for inducing bNAb responses against a wide range of infectious diseases.
Advisor
Bar, Katharine, J