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Principal Investigator: CHRISTOPHER C BRODER
Organization: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
Fiscal Year: 2019
Award: $193,928
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract
The emergence of pathogenic viruses represents continuous infectious disease threats to public health.
Among these, the paramyxoviruses, which include many important human and animal pathogens, also include
two excellent examples of emerged, zoonotic pathogens of importance: the henipaviruses; Hendra virus (HeV)
and Nipah virus (NiV). HeV and NiV have a uniquely broad host tropism capable of infecting at least 18 animal
species across 6 orders of mammals. HeV and NiV can also cause a systemic and often fatal respiratory
and/or neurological disease in 11 mammalian species including humans. These henipaviruses remain
significant biothreats to humans and economically important livestock in Australia and throughout South East
Asia, and there are no vaccines or antivirals approved for human use. The henipaviruses are single-stranded,
negative sense, enveloped RNA viruses and possess two membrane anchored glycoproteins involved in virus
entry, one for host cell receptor attachment (G glycoprotein) and the other a fusion (F) glycoprotein which
facilitates virion and host cell membrane fusion. The G and F glycoproteins are the major antigenic targets of
neutralizing antibodies and also the focus of several vaccine strategies. In contrast however, antiviral drug
discovery for HeV and NiV has been significantly hampered because of the requirements of biosafety level-4
(BSL-4) containment, and presently there is a complete lack of any effective antiviral therapeutics against
henipaviruses for human use. We have been extensively characterizing the recently identified non-pathogenic
species of henipavirus, Cedar virus (CedPV). Using recombinant viral glycoprotein mediated cell-cell fusion
assays, we have discovered that CedPV-mediated membrane fusion is similar to HeV and NiV, however, the
ephrin receptor tropism of CedPV was found to be remarkably broad and fusion could be triggered by ephrin-
B1 and -B2 as well as the glycophosphatidylinositol-anchored A subtype ephrins-A1, -A2, and -A5. The cell-
cell fusion activity supported by each of these ephrin receptors also correlated with CedPV G binding ability as
measured by surface plasmon resonance. Further, we have recently rescued recombinant CedPV encoding
eGFP (rCedPV-GFP) using a reverse genetics approach. Our rCedPV platform represents a new virological
system that can be used in a variety of applications to study various aspects of henipavirus cell biology and
host cell interactions safely under BSL-2 containment. But of further importance, it is also an authentic
henipavirus infection and replication reporter system now suitable for high-throughput screening (HTS) for the
discovery of potentially pan-anti-henipavirus molecules. Using this new live-henipavirus platform, our
objectives here will be to develop and optimize a rCedPV reporter system and utilize it in an HTS assay to
discover antiviral molecules capable of inhibiting the infection of henipaviruses. Specifically, we will: 1) Adapt
and develop CedPV reporters for the HTS assay; 2) Optimize the HTS parameters of recombinant virus
infection and reporter activities; and 3) Pilot a HTS assay using the Prestwick Chemical Library.
Terms: <Animals><Antiviral Agents><Antiviral Drugs><Antivirals><Assay><Australia><Binding><Bioassay><Biologic Assays><Biological Assay><Cek5 Ligand><Cek5 RPTK Ligand><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Culture Techniques><Cell Interaction><Cell Line><Cell Signaling><Cell fusion><Cell membrane><Cell-to-Cell Interaction><CellLine><Cells><Cellular biology><Chicago><Collaborations><Communicable Diseases><Containment><Cytoplasmic Membrane><EFNB1 Gene Product><EPLG1><East Asia><Eastern Asia><Eck Ligand><Eck RPTK Ligand><Elk-L Protein><Endothelium Secreted Protein B61><Eph Family Receptor Interacting Protein B1><Eph Family Receptors><Eph Receptor Tyrosine Kinase><Eph Receptors><Ephrin Receptors><Ephrin-A1><Ephrin-B1><Epl1 Protein><Equine Morbillivirus><Far East><Farm Animal><General Viruses><Generalized Growth><Glycoproteins><Growth><Hendra><Hendra Virus><Henipavirus><Henipavirus Infections><High Throughput Assay><Human><Illinois><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Intracellular Communication and Signaling><Kinetics><LERK-1 Protein><LERK-2 Protein><Lead><Libraries><Livestock><Luc Gene><Luciferase Gene><Luciferase Immunologic><Luciferases><Lung diseases><Mammalia><Mammals><Measures><Mediating><Medical><Membrane><Membrane Fusion><Modern Man><Molecular Interaction><Nervous System Diseases><Neurologic Disorders><Neurological Disorders><Nipah Virus><Paramyxoviridae><Paramyxovirus><Pathogenicity><Pb element><Plasma Membrane><Public Health><Pulmonary Diseases><Pulmonary Disorder><RNA Viruses><Receptor Cell><Recombinants><Reporter><Reporter Genes><Research><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><Series><Signal Transduction><Signal Transduction Systems><Signaling><Strains Cell Lines><Surface Plasmon Resonance><System><Testing><Texas><Therapeutic><Time><Tissue Growth><Tropism><Universities><Vaccines><Viral><Viral Diseases><Virion><Virus><Virus Diseases><Virus Particle><Virus Replication><Work><Zoonoses><Zoonotic><Zoonotic Infection><anti-viral agents><anti-viral drugs><anti-virals><base><bio-threat><biological signal transduction><biothreat><cell biology><cell culture><chemical library><cultured cell line><disease of the lung><disorder of the lung><drug discovery><elk Ligand><experiment><experimental research><experimental study><genetic approach><glycoprotein G><heavy metal Pb><heavy metal lead><high throughput screening><inhibitor><inhibitor/antagonist><insight><lung disorder><membrane structure><nervous system disorder><neurological disease><neutralizing antibody><ontogeny><pathogen><pathogenic virus><plasmalemma><recombinant virus><reverse genetics><screening><small molecule><small molecule libraries><viral infection><viral multiplication><viral pathogen><viral replication><virology><virus infection><virus multiplication><virus-induced disease>