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Principal Investigator: ANDREW HAYHURST
Organization: TEXAS BIOMEDICAL RESEARCH INSTITUTE
Fiscal Year: 2023
Award: $297,000
Funding agency: National Institute of Allergy and Infectious Diseases
ABSTRACT
The current coronavirus (CoV) pandemic, seasonal infections by other CoV and other “cold” viruses,
plus the need for annual influenza vaccinations exemplify the challenges posed by viral antigenic drift and shift.
Targeting landscapes of viral structural proteins displayed on the surfaces of virus particles and or the surfaces
of infected cells has been the primary basis for developing antibody-based therapeutics. Although great
advances have been made in trying to identify regions of these surface displayed proteins that are conserved
and less prone to “escape” antibody binding, it appears to be a continual battle of cat and mouse as we are
seeing with continual emergence of CoV “Variants of Concern”. In contrast, viral structural proteins that remain
inside virus particles and cells, avoid the impact of cyclical antibody selection, and tend to be far more
conserved. Oligomeric assemblies of these proteins can also blunt the impact of antiviral escape mutations
owing to the mix of mutant and wild-type monomers present in the parent cell. Furthermore, the stoichiometry
required of an antiviral to impede oligomer function need not be necessarily 1 antiviral to 1 monomer since,
especially if the antiviral were a crosslinker, its impact would be relayed beyond the immediate contact to
neighboring oligomers. Our long-term hypothesis is that affinity reagents capable of binding an internal
oligomeric structural protein of all species of a viral genus uniformly will impede viral assembly when present
as dimeric crosslinkers in a manner that is both forward capable and has much reduced susceptibility to viral
escape. We will explore the antiviral potential of dimeric crosslinkers using viruses of the genus Ebolavirus as
our model and a novel, rare nanobody manifesting uniform reactivity to nucleoprotein of all 6 species. Our two
specific aims are: (1) we will engineer mammalian cell expression vectors encoding nanobody homodimers
and assess antiviral activity using virus like particle surrogates at BSL-2 following plasmid transfection to drive
intrabody expression, (2) we will engineer E. coli expression vectors encoding nanobody homodimers fused to
cell penetrating peptides and glycosaminoglycan binding motifs and assess antiviral activity following protein
transduction of virus infected cells at BSL-4. Success will demonstrate a novel antiviral strategy that can then
be thoroughly explored for the propensity to select escape mutants relative to an existing neutralizing antibody
regime to test whether the strategy is more “escape-proof”. The overall approach should be applicable to other
human viral pathogens by carefully retuning the affinity reagent, with adequate time and resources, to
maximize broad long-term impact in helping to safe guard human health.
Terms: <Affinity><Antibodies><Antibody Therapy><Antigen Targeting><Antigenic Determinants><Area><Avidity><Binding><Binding Determinants><Capsid><Cats><Cats Mammals><Cell Body><Cells><CoV emergence><CoV pandemic><Compensation><Coronaviridae><Coronavirus><Crosslinker><DNA><Deoxyribonucleic Acid><Detection><Development><Diagnostic><Disease><Disorder><Domestic Cats><Drugs><E coli><E. coli><EBOV><Ebola virus><Ebola-like Viruses><Engineering><Ensure><Envelope Protein><Epitopes><Escape Mutant><Escherichia coli><Evolution><Exhibits><Family><Feline Species><Felis catus><Felis domestica><Felis domesticus><Felis sylvestris catus><Flu vaccination><Foundations><Future><Generations><Genes><Genetic Alteration><Genetic Change><Genetic defect><Glycosaminoglycans><Goals><Health><Human><Immune system><In Vitro><Individual><Infection><Influenza immunization><Influenza vaccination><Intervention><Intervention Strategies><Intrabody><Mammalian Cell><Mediating><Medication><Mice><Mice Mammals><Modeling><Modern Man><Molecular Interaction><Monitor><Mucopolysaccharides><Murine><Mus><Mutation><Nature><Nucleoproteins><Parents><Penetration><Peptides><Pharmaceutic Preparations><Pharmaceutical Preparations><Plasmids><Polymers><Predisposition><Process><Prophylactic vaccination against influenza><Proteins><RNA Viruses><RNA vaccine><RNA-based vaccine><Reagent><Research><Research Resources><Resources><Risk><Seasons><Short interfering RNA><Site><Small Interfering RNA><Specificity><Structural Protein><Surface><Susceptibility><System><Technology><Testing><Therapeutic antibodies><Time><Transfection><Viral><Viral Activity><Viral Function><Viral Gene Products><Viral Gene Proteins><Viral Genome><Viral Hemorrhagic Fevers><Viral Physiology><Viral Proteins><Viral Structural Proteins><Virion><Virus><Virus Assembly><Virus Cultivation><Virus Particle><Virus Replication><Virus-like particle><Work><antibody based therapies><antibody treatment><antibody-based therapeutics><antibody-based treatment><corona virus><corona virus emergence><coronavirus emergence><coronavirus pandemic><crosslink><design><designing><developmental><dimer><drug/agent><ebolavirus><emergent CoV><emergent corona virus><emergent coronavirus><emerging CoV><emerging corona virus><emerging coronavirus><env Antigens><env Gene Products><env Polyproteins><env Protein><expression vector><flu immunisation><genome mutation><hemorrhagic fever><human disease><influenza virus vaccination><interest><interventional strategy><mRNA vaccine><mRNA-based vaccine><member><monomer><mutant><nCoV><nanobodies><nanobody><neutralizing antibody><new CoV><new approaches><new corona virus><new coronavirus><non-human primate><nonhuman primate><novel><novel CoV><novel approaches><novel corona virus><novel coronavirus><novel strategies><novel strategy><parent><particle><pathogenic virus><polymer><polymeric><preference><protein oligomer><sdAb><siRNA><single domain antibodies><stoichiometry><success><tissue culture><vaccination against influenza><variants of concern><viral assembly><viral multiplication><viral pathogen><viral replication><virus genome><virus multiplication><virus pathogen><virus protein><virus-like nanoparticles><viruslike particle>