A DNA nanotechnology vaccine platform for delivery and translation of HIV-1 1086C gp160 Env LNP

PROJECT SUMMARY ABSTRACT The human immune response to HIVis complex with the virus demonstrating remarkable evasion mechanismsSignificant progress has recently been made in HIV vaccine development based on early stage clinical trials and subsequent investigations into the nature of protective immunitySeveral promising messenger RNAmRNAbased HIV vaccines that target the HIV envelope proteinEnvhave been developed and tested with promising results in animal and nonhuman primate modelsThe vaccines are based on lipid nanoparticleLNPtechnology which relies on the LNP lipid bilayer to protect the mRNA cargo and promote cellular uptake but lacks the specificity to traffic the vaccine to dendritic cellsDCand secondary lymphoid organsa deficiency addressed through depotintradermal or subcutaneous injectionRather than modifying the lipid bilayer of an mRNA LNP vaccine to impart desirable physical and biological attributeswe propose a more versatile approachdecoupling vaccine functionality into layers that address extracellular and intracellular issues and goals separatelyToward this endwe propose to design and develop a customizedspherical DNA nanostructureorigamiconstruct to encapsulate an existing mRNA LNP vaccine with the goals of increasing immunogenicity and prolonging specific antibody productionThe origami will serveas an anionic shell coated with ligands for DC targetingas an anchor for co delivery of adjuvantsandas a protective layer against enzyme degradation and nonspecific cellular interactionsWe will develop and optimize protocols for synthesizing the origami LNP complex and characterize it using atomic force microscopydynamic light scatteringfluorescence microscopyand gel chromatographyWe will then demonstrate the capacity of the resulting construct to transfect and effect expression of Env in cell cultures Env protein by western blotTargeting and uptake will be assessed using labeled DNA constructs and fluorescence microscopyFinallywe will evaluate vaccine efficacy to stimulate a specific humoral immune response in a murine model by quantifying antibody titers by ELISA assayIf successfulthe proposed origami LNP approach to nanocarrier design could shift current drug delivery practice not only for mRNAbased vaccinesbut for a wide variety of therapeutic applications PROJECT NARRATIVE There is an urgent need for an effective vaccine against HIV AIDS to protect and treat millions of sufferers and stop the ongoing epidemicTowards this end significant progress has been madeand we propose to improve the performance of a promising HIVvaccine candidate by enclosing it in a customized DNA nanostructure tailored to enhance the immune response