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Principal Investigator: Katherine W Ferrara
Organization: STANFORD UNIVERSITY
Fiscal Year: 2020
Award: $655,422
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Abstract of: Proposed COVID-19-related supplement to
In vivo PET imaging of novel engineered AAVs informs capsid design
There is an urgent need to develop tools to assess viral pathogenesis and the efficacy of potential therapeutics
for novel viruses such as SARS-CoV-2. While organoid and cell-based assays have been broadly used to
assess the candidate receptor of such viruses, these assays cannot answer key questions as to: 1) whether
multiple receptors for the virus exist, 2) the in vivo receptor affinity of the virus and accumulation within the
upper respiratory tract, 3) transport of the virus into the vascular system and ultimately to the heart and
kidneys, and 4) the resulting transfection of these various sites. Our laboratory has previous developed
methods to label adeno-associated viruses and track their transport following systemic injection. We found
that these engineered viruses carry cargo attached to the capsid across the blood brain barrier and the cargo
accumulates deep within the brain. Combined optical and PET studies have suggested that binding of the
virus to its receptor results in transcytosis of the intact capsid. We hypothesize that coronaviruses may
possess similar capabilities to be transported across the lung epithelium. We plan to address key issues by
assessing receptor binding and transduction using PET and optical imaging. Our resulting specific aims are the
following: 1) development and validation of tagging strategies to image pseudotype viral particles at BSL2, 2)
development and validation of reporter gene strategies to image transduction of engineered viruses, and 3)
application and dissemination of these dual strategies to assess viral transport, transduction and susceptibility
to available therapies including a) antibodies, b) protease inhibitors, and c) fusion inhibitors. We propose to
develop and image engineered viruses expressing the spike protein and a reporter gene and track these
viruses within a model of lung fibrosis and a mouse model with a humanized ACE2 receptor. We will leverage
the capabilities to label and track viral capsids and transduction developed within this R01 and key capabilities
of Stanford University. Pseudotype viruses based on vesicular stomatitis virus (VSV) and lentivirus have been
developed with spike proteins corresponding to SARS-CoV or SARS-CoV-2. Further, replicons with intact viral
proteases have been engineered. We propose to collaborate with those developing and testing engineered
viruses and therapeutics at Stanford, including Jan Carette and Catherine Blish in addition to the key personnel
on our parent project. At the conclusion of each phase of this project, we will disseminate strategies for the
incorporation of a PET tag, a reporter gene, and dual PET imaging protocols. We hypothesize that these tools
can be disseminated and rapidly modified to assess both SARS-CoV-2 and future viruses. We will make our
technology available through commercial and scientific partners.
Terms: <2019 novel coronavirus><2019-nCoV><Address><Adeno-Associated Viruses><Affinity><Amino Acids><Angiotensin Converting Enzyme><Angiotensin I-Converting Enzyme><Antibodies><Antiproteases><Area><Assay><Binding><Bioassay><Biologic Assays><Biological Assay><Blood - brain barrier anatomy><Blood Serum><Blood Vessels><Blood-Brain Barrier><Brain><Brain Nervous System><CD143 Antigens><COVID-19><COVID19><Capsid><Capsid Proteins><Carboxycathepsin><Cardiac><Cell Body><Cells><Chemicals><Clinical><Clinical Treatment Moab><Coat Proteins><Coronaviridae><Coronavirus><Data Bases><Databases><Dependoparvovirus><Dependovirus><Development><Dipeptidyl Peptidase A><Drug Kinetics><EBOV><Ebola virus><Encephalon><Endopeptidase Inhibitors><Engineering><Epithelial><Epitheliasin Gene><Epithelium><Epithelium Part><Esteroproteases><Feces><Future><Heart><Hemato-Encephalic Barrier><High Prevalence><Host Factor><Host Factor Protein><Human><Human Resources><Hypertension><Image><Infection><Injections><Integration Host Factors><Kidney><Kidney Urinary System><Kininase A><Kininase II><Label><Laboratories><Lentivirinae><Lentivirus><Light><Lung><Lung Respiratory System><Manpower><Mediating><Membrane Fusion><Methods><Modeling><Modern Man><Modification><Molecular Interaction><Monoclonal Antibodies><Nucleic Acids><Optics><Organ><Organoids><PET><PET Scan><PET imaging><PETSCAN><PETT><PRSS10><Pathology><Peptidase Inhibitors><Peptidases><Peptide Hydrolase Inhibitors><Peptide Hydrolases><Peptide Peptidohydrolase Inhibitors><Peptidyl-Dipeptidase A><Pharmacokinetics><Phase><Photoradiation><Positron Emission Tomography Medical Imaging><Positron Emission Tomography Scan><Positron-Emission Tomography><Predisposition><Protease Antagonists><Protease Gene><Protease Inhibitor><Proteases><Proteinase Inhibitors><Proteinases><Proteins><Proteolytic Enzymes><Protocol><Protocols documentation><Pulmonary Fibrosis><Rad.-PET><Receptor Protein><Replication Unit><Replicon><Reporter Genes><Reporting><SARS><SARS Virus><SARS corona virus><SARS coronavirus><SARS coronavirus disease><SARS-Associated Coronavirus><SARS-CoV><SARS-CoV disease><SARS-CoV-2><SARS-CoV2><SARS-Related Coronavirus><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related coronavirus 2><Serine Endopeptidases><Serine Protease><Serine Protein Hydrolases><Serine Proteinases><Serum><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe Acute Respiratory Syndrome coronavirus disease><Severe acute respiratory syndrome coronavirus 2><Site><Subfamily lentivirinae><Surface><Susceptibility><T-Cells><T-Lymphocyte><TMPRSS2><TMPRSS2 gene><Technology><Testing><Therapeutic><Time><Training><Transfection><Universities><Upper respiratory tract><VSV><Validation><Vascular Hypertensive Disease><Vascular Hypertensive Disorder><Vascular System><Vesicular Stomatitis Virus><Vesicular stomatitis Indiana virus><Viral><Viral Burden><Viral Coat Proteins><Viral Load><Viral Load result><Viral Outer Coat Protein><Viral Pathogenesis><Viral Receptor><Viremia><Virus><Virus Receptors><Virus Replication><Virus-Lenti><Wuhan coronavirus><adeno associated virus group><aminoacid><base><bloodbrain barrier><co-morbid><co-morbidity><comorbidity><corona virus><corona virus disease 2019><coronavirus disease 2019><data base><design><designing><develop a vaccine><development of a vaccine><developmental><drug development><ebolavirus><high blood pressure><hyperpiesia><hyperpiesis><hypertensive disease><image-based method><imaging><imaging method><imaging modality><in vivo><inhibitor><inhibitor/antagonist><lung fibrosis><mAbs><mouse model><murine model><novel><novel virus><optic imaging><optical><optical imaging><parent project><particle><personnel><positron emission tomographic (PET) imaging><positron emission tomographic imaging><positron emitting tomography><pulmonary><receptor><receptor binding><receptor bound><renal><severe acute respiratory syndrome-CoV><stool><thymus derived lymphocyte><tool><transcytosis><vaccine development><vaccine formulation><vascular><viraemia><viral multiplication><viral replication><viral sepsis><virus multiplication><virus pathogenesis><virusemia>