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Principal Investigator: Jose Alfonso Rodriguez
Organization: UNIVERSITY OF CALIFORNIA LOS ANGELES
Fiscal Year: 2019
Award: $234,000
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
New World Hemorrhagic Fever (NWHF) mammarenaviruses are category A pathogens that
pose a grave threat to humans due their high infectivity, mortality between 15-30% when
untreated, and our lack of therapeutic options. A subgroup of NWHF mammarenaviruses
include the clade B pathogens Junin, Machupo, Guanarito, Chapare and Sabia, which
collectively put thousands at risk per year and are considered potential bioweapons. Infection of
human cells by New World hemorrhagic fever viruses is mediated by the binding of their surface
glycoprotein (GP1) to the human Transferrin Receptor 1 (hTfR1/CD71). However, while all viral
glycoproteins bind the same receptor, structural variability among GP1 proteins curtails efforts
to develop broadly neutralizing therapeutics against the entire group of viruses; this is one of the
most complex challenges faced by targeted therapies. We recently reported the use of a
monoclonal antibody (ch128.1), which targets the ectodomain of hTfR1, to efficiently inhibit the
infection of mammalian cells by pseudovirus expressing pathogenic NWHF GP1. We aim to
develop an effective and broadly neutralizing therapeutic against NWHF mammarenaviruses
and establish the structural foundation for its inhibition. To achieve this goal, we will generate
and characterize single chain antibody fragments (scFv and scFab) with the variable region of
the monoclonal antibody 128.1 that prevent binding of all pathogenic NWHF GP1 proteins to
hTfR1. We will recombinantly produce and purify the single chain fragments and biochemically
confirm their expected high affinity and specificity for hTfR1. We will also evaluate fragments for
their ability to inhibit entry of pseudoviruses decorated with NWHF virus GP1 into cells
expressing hTfR1. We will then crystallize our fragments alone and bound to hTfR1, and
determine their atomic structure. Our elucidation of these structures will benefit from frontier
crystallographic tools at our disposal, including microfocal x-ray diffraction and electron micro-
diffraction (MicroED). The resulting structures will provide us with a template from which we can
generate new generations of improved and broadly acting therapeutics against all clade B
NWHF mammarenaviruses.
Terms: <7S Gamma Globulin><Ab-dependent cellular cytotoxicity><Affinity><Antibodies><Antibody Fragments><Antigen Binding Fragment><Arenaviridae><Arenaviridae Infections><Arenavirus><Arenavirus Infections><Arenavirus group><Argentine hemorrhagic fever virus><Argentinian Hemorrhagic Fever Virus><Autoimmune><Autoimmune Process><Binding><Biochemical><Bolivian Hemorrhagic Fever Virus><CD71><Capsid Proteins><Category A pathogen><Category A priority pathogen><Cause of Death><Cell Body><Cell Surface Glycoproteins><Cell Surface Proteins><Cell surface><Cells><Chapare><Chapare Virus><Circulatory Collapse><Clinical Research><Clinical Study><Clinical Treatment Moab><Coat Proteins><Coma><Comatose><Communicable Diseases><Complement-Dependent Cytotoxicity><Complex><Constant Region><Crystallization><Development><Disease><Disorder><Dose><Dysfunction><Electrons><Endocytosis><Evaluation><External Domain><Extracellular Domain><FDA approved><Fab Fragments><Fab Immunoglobulins><Foundations><Functional disorder><Future><GP1><GTP-Binding Protein 1><GTPBP1><GTPBP1 gene><General Viruses><Generations><Genetic Alteration><Genetic Change><Genetic defect><Genetics-Mutagenesis><Glycoproteins><Goals><Guanarito><Guanarito virus><Human><Ig Constant Region><Ig Variable Region><IgG><IgG1><Immunoglobulin Constant Region><Immunoglobulin Fragments><Immunoglobulin G><Immunoglobulin V><Immunoglobulin Variable Region><Immunoglobulin, F(ab) Fragment><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Junin><Junin virus><Length><Light><Machupo><Machupo virus><Mammalian Cell><Mediating><Membrane Glycoproteins><Methods><Micro Electron Diffraction><MicroEd><Modeling><Modern Man><Molecular><Molecular Interaction><Molecular Target><Monoclonal Antibodies><Mutagenesis><Mutagenesis Molecular Biology><Mutation><NIAID><National Institute of Allergy and Infectious Disease><Negative Beta Particle><Negatrons><Neurologic Manifestations><Neurologic Signs and Symptoms><Neurologic Symptoms><Neurological Manifestations><Neurological Signs and Symptoms><Pathogenicity><Photoradiation><Physiopathology><Production><Property><Proteins><Reaction><Receptor Protein><Recombinant Antibody><Recombinants><Reporting><Retroviridae><Retroviruses><Risk><Seizures><Shock><Specificity><Structure><Subgroup><Surface><Surface Glycoproteins><Symptoms><System><TFR gene><TFR protein><TFR1><TFRC><TFRC gene><TRFR><Therapeutic><Therapeutic Uses><Transferrin Receptor><Transferrin Receptor 1><Treatment Efficacy><Variable Region><Venezuelan hemorrhagic fever virus><Viral><Viral Coat Proteins><Viral Hemorrhagic Fevers><Viral Outer Coat Protein><Virus><Virus Inhibitors><Virus-Retrovirus><X ray diffraction><X ray diffraction analysis><antibody dependent cell mediated cytotoxicity><antibody dependent cytotoxicity><antibody-dependent cell cytotoxicity><antibody-dependent cellular cytotoxicity><antibody-mediated cytotoxicity><biological weapon><bioweapon><circulatory shock><cost><cross-link><crosslink><developmental><effective therapy><effective treatment><flu><frontier><genome mutation><hemorrhagic fever><hemorrhagic fever virus><improved><innovate><innovation><innovative><intervention efficacy><mAbs><mortality><neural manifestation><neutralizing antibody><new drug treatments><new drugs><new therapeutics><new therapy><next generation therapeutics><novel><novel drug treatments><novel drugs><novel therapeutics><novel therapy><pathogen><pathophysiology><pre-clinical study><preclinical study><prevent><preventing><receptor><side effect><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic efficacy><therapeutically effective><therapy efficacy><tool><viral inhibitor>