Document text
Principal Investigator: David Lawrence Danley
Organization: PARABON NANOLABS, INC.
Fiscal Year: 2019
Award: $225,220
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY/ABSTRACT
The human immune response to HIV-1 is complex with the virus demonstrating remarkable evasion
mechanisms. Significant progress has recently been made in HIV vaccine development based on early-stage
clinical trials and subsequent investigations into the nature of protective immunity. Several promising
messenger RNA (mRNA)-based HIV vaccines that target the HIV envelope protein (Env) have been developed
and tested with promising results in animal and nonhuman primate models. The vaccines are based on lipid
nanoparticle (LNP) technology 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 cells (DC) and secondary lymphoid
organs – a deficiency addressed through depot, intradermal or subcutaneous injection.
Rather than modifying the lipid bilayer of an mRNA-LNP vaccine to impart desirable physical and biological
attributes, we propose a more versatile approach: decoupling vaccine functionality into layers that address
extracellular and intracellular issues and goals separately. Toward this end, we propose to design and develop
a customized, spherical DNA nanostructure (origami) construct to encapsulate an existing mRNA-LNP vaccine
with the goals of increasing immunogenicity and prolonging specific antibody production. The origami will
serve (1) as an anionic shell coated with ligands for DC targeting, (2) as an anchor for co-delivery of adjuvants,
and (3) as a protective layer against enzyme degradation and nonspecific cellular interactions.
We will develop and optimize protocols for synthesizing the origami-LNP complex and characterize it using
atomic force microscopy, dynamic light scattering, fluorescence microscopy, and gel chromatography. We will
then demonstrate the capacity of the resulting construct to transfect and effect expression of Env in cell
cultures Env protein by western blot. Targeting and uptake will be assessed using labeled DNA constructs and
fluorescence microscopy. Finally, we will evaluate vaccine efficacy to stimulate a specific humoral immune
response in a murine model by quantifying antibody titers by ELISA assay. If successful, the proposed
origami-LNP approach to nanocarrier design could shift current drug delivery practice not only for mRNA-
based vaccines, but for a wide variety of therapeutic applications.
Terms: <AIDS/HIV><AIDS/HIV problem><ATGN><Ab response><Address><Adjuvant><Animals><Antibody Formation><Antibody Production><Antibody Response><Antibody titer measurement><Antigens><Area><Assay><Atomic Force Microscopy><BALB C Mouse><BALB/c><Bioassay><Biologic Assays><Biological><Biological Assay><Caliber><Cell Body><Cell Culture Techniques><Cells><Charge><Chitosan><Complex><Computer-Aided Design><Computer-Assisted Design><Culture Media><Custom><Cyclic GMP><Cytosine Polynucleotides><DNA><Data><Dendritic Cells><Dendritic cell activation><Deoxyribonucleic Acid><Designing computer software><Diameter><Drug Delivery><Drug Delivery Systems><Drug Targeting><ELISA><Early-Stage Clinical Trials><Elements><Encapsulated><Envelope Protein><Enzyme Gene><Enzyme-Linked Immunosorbent Assay><Enzymes><Epidemic><Exclusion Chromatography><Fluorescence Light Microscopy><Fluorescence Microscopy><Fluorometry><Force Microscopy><Formulation><Gel Chromatography><Gel Filtration><Gel Filtration Chromatography><Gel Permeation Chromatography><General Viruses><Germinal Center><Goals><Guanosine Cyclic Monophosphate><HIV envelope protein><HIV vaccine><HIV-1><HIV-1 vaccine><HIV-I><HIV/AIDS><HIV/AIDS Vaccines><HIV/AIDS problem><HIV1><HIV1 vaccine><Human><Human Immunodeficiency Virus Type 1><Human immunodeficiency virus 1><Hydrophobic Surfaces><Immune response><Immunity><Immunological response><In Vitro><Inbred BALB C Mice><Incubated><Interpersonal Communication><Investigation><Laboratories><Legal patent><Ligands><Lipid Bilayers><Lipids><Luciferase Immunologic><Luciferases><Lymphoid><Messenger RNA><Modeling><Modern Man><Molecular Weight><Nanostructures><Nanotechnology><Nature><Non-Polyadenylated RNA><Oligo><Oligonucleotides><Organ><Patents><Performance><Personal Communication><Phase><Phase 1 Clinical Trials><Phase I Clinical Trials><Poliglusam><Poly C><Polycytidylic Acids><Production><Proteins><Protocol><Protocols documentation><RNA><RNA Gene Products><Ribonucleic Acid><Scanning Force Microscopy><Shapes><Software Design><Specificity><Structure><Structure of germinal center of lymph node><Subcutaneous Injections><Surface><System><Technology><Testing><Therapeutic><Transfection><Translations><Universities><Vaccines><Variant><Variation><Veiled Cells><Virus><Western Blotting><Western Immunoblotting><Work><Yang><antibody biosynthesis><antibody titering><base><cGMP><cell culture><design><designing><develop a vaccine><development of a vaccine><drug development><env Antigens><env Gene Products><env Polyproteins><env Protein><extracellular><gp160><growth media><homopolymer 5'-Cytidylic acid><host response><human immunodeficiency virus vaccine><immunogen><immunogenicity><immunoglobulin biosynthesis><immunoresponse><improved><in vivo><intradermal injection><light scattering><lipid bilayer membrane><lipid nanoparticle><mRNA><mouse model><murine model><nano carrier><nano meter scale><nano meter sized><nano particle delivery><nano scale><nano tech><nano technology><nano-structures><nano-technological><nanocarrier><nanometer scale><nanometer sized><nanoparticle delivered><nanoparticle delivery><nanoscale><nanotech><nanotechnological><neutralizing antibody><new vaccines><next generation vaccines><non-human primate><nonhuman primate><novel vaccines><off-label application><off-label prescribing><off-label use><off-patent><oligos><phase 1 trial><phase I protocol><protein blotting><protein expression><response><uptake><vaccine candidate><vaccine delivery><vaccine development><vaccine efficacy><vaccine formulation>