Document text
Principal Investigator: Harvey Bruce Pollard
Organization: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
Fiscal Year: 2023
Award: $663,601
Funding agency: National Heart Lung and Blood Institute
Abstract
Over 860,000 COVID-19 deaths have occurred in the U.S., and more than 5.5 million deaths have
occurred world-wide. Consequently, development of effective antiviral drugs that block infectivity and airway
inflammation continue to be a global health priority. Inflammation in the COVID-19 airway is due to increased
NFκB and Epithelial Sodium channel (ENaC) signaling. Cystic fibrosis (CF), a genetic disease caused by
inactivating mutations in the CFTR gene, also has the same proinflammatory NFκB and ENaC signaling
phenotype in the airway. Using a differentiated human epithelial "lung-on-a-chip" platform, we find that ACE2,
the receptor protein for the SARS-CoV-2 Spike protein, co-immunoprecipitates with CFTR. Furthermore,
exposure of differentiated epithelia to the SARS-COV-2 Spike protein dose-dependently suppresses cyclic
AMP-dependent CFTR chloride channel activity and CFTR protein expression. Spike-dependent loss of CFTR
also activates TRADD-dependent NFκB signaling and proteolytically activates ENaC. We have also found that
Spike-dependent loss of CFTR may be due to failure of endosomal recycling to return apical CFTR to the
plasma membrane. Finally, we find that nanoMolar concentrations of cardiac glycoside drugs such as
ouabain, digitoxin and digoxin, which competitively inhibit Spike:ACE2 binding, rescue Spike-dependent
reduction in CFTR activities. We have therefore hypothesized that binding of SARS-CoV-2 Spike protein to
ACE2 in the lung causes loss of CFTR signaling and activation of proinflammatory NFκB and ENaC
signaling. To further test this hypothesis we propose the following Specific Aims: SA #1: To define the
mechanism by which SARS-CoV-2 Spike protein reduces CFTR channel activity and CFTR protein
levels. We will determine the mechanism by which ACE2 interacts with CFTR. We will determine the
mechanism by which Spike protein interaction with ACE2 leads to loss of CFTR. SA#2: To define the
mechanism by which SARS-CoV-2 Spike protein drives activation of ENaC and NFκB signaling. We will
determine the mechanism by which Spike-induced loss of CFTR protein results in activation of TRADD-
dependent NFκB ENaC signaling. SA#3: To identify protective mechanisms of cardiac glycosides on cell
and animal models of COVID-19 disease. We will test whether cardiac glycoside drugs block native SARS-
CoV-2 infection of epithelia and rescue hamster models of COVID-19. .
Novelty and Significance: To our knowledge this is the first time COVID-19 airway inflammation has been
traced to inhibition of CFTR signaling. Consistently, COVID-19 patients who are also CF carriers, with only one
wildtype CFTR gene and only 50% of CFTR function, were recently reported to suffer more severe COVID-19
symptoms and earlier death than normal subjects with COVID-19.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><3'5'-cyclic ester of AMP><ACE2><Acocantherin><Acolongifloroside K><Adenosine Cyclic 3',5'-Monophosphate><Adenosine Cyclic Monophosphate><Adenosine, cyclic 3',5'-(hydrogen phosphate)><Affect><Air><Animal Model><Animal Models and Related Studies><Antiviral Agents><Antiviral Drugs><Antivirals><Apical><Automobile Driving><Basal Cell><Binding><Biopsy><CFTR><CFTR Protein><COVID disease severity><COVID infected patient><COVID patient><COVID positive patient><COVID severity><COVID-19><COVID-19 S protein><COVID-19 associated death><COVID-19 associated fatality><COVID-19 associated mortality><COVID-19 death><COVID-19 disease severity><COVID-19 fatality><COVID-19 induced death><COVID-19 induced fatality><COVID-19 induced mortality><COVID-19 infected patient><COVID-19 infection><COVID-19 mortality><COVID-19 patient><COVID-19 positive patient><COVID-19 related death><COVID-19 related fatality><COVID-19 related mortality><COVID-19 severity><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 virus><COVID-19 virus infection><COVID19><COVID19 S protein><COVID19 associated death><COVID19 associated fatality><COVID19 associated mortality><COVID19 death><COVID19 disease severity><COVID19 fatality><COVID19 induced death><COVID19 induced fatality><COVID19 induced mortality><COVID19 infection><COVID19 mortality><COVID19 patient><COVID19 positive patient><COVID19 related death><COVID19 related fatality><COVID19 related mortality><COVID19 severity><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><CV-19><CV19><Cardiac Glycosides><Cardiotonic Steroids><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell membrane><Cell model><Cells><Cellular model><Cessation of life><Chloride Channels><Chloride Ion Channels><Co-Immunoprecipitations><CoV-2><CoV2><Cricetinae><Cyclic AMP><Cystic Fibrosis><Cystic Fibrosis Transmembrane Conductance Regulator><Cytoplasmic Membrane><Data><Death><Development><Digitoxin><Digoxin><Disease><Disorder><Dose><Drugs><ENaC><Endosomes><Epithelial Cells><Epithelium><Failure><G-Strophanthin><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic defect><Hamsters><Hamsters Mammals><Health Priorities><Histologic><Histologically><Human><Incubated><Infection><Inflammation><Inflammatory><Inflammatory Response><Intracellular Communication and Signaling><Lanoxin><Liquid substance><Lung><Lung Respiratory System><Medication><Modeling><Modern Man><Molecular Interaction><Mucoviscidosis><Mutation><Organ><Ouabain><Penetration><Pharmaceutic Preparations><Pharmaceutical Preparations><Phenotype><Plasma Membrane><Process><Progenitor Cells><Proteins><RNA analysis><Receptor Protein><Receptosomes><Recovery><Recycling><Regulator Genes><Reporting><Reproducibility><Respiratory Epithelium><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 associated death><SARS-CoV-2 associated fatality><SARS-CoV-2 associated mortality><SARS-CoV-2 death><SARS-CoV-2 disease severity><SARS-CoV-2 fatality><SARS-CoV-2 induced death><SARS-CoV-2 induced fatality><SARS-CoV-2 induced mortality><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 inhibitor><SARS-CoV-2 mortality><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 related death><SARS-CoV-2 related fatality><SARS-CoV-2 related mortality><SARS-CoV-2 severity><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 infection><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Secretory Cell><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Signal Transduction><Signal Transduction Systems><Signaling><Structure of respiratory epithelium><Symptoms><TNF Receptor-Associated Death Domain Protein><TNF Receptor-Associated Protein with a Death Domain><TNFR1-Associated Death Domain Protein><TNFRSF1A-Associated via Death Domain><TRADD><TRADD gene><Telomerase><Testing><Time><Transcriptional Regulatory Elements><Viral><Virus><Wuhan coronavirus><adenosine 3'5' monophosphate><airway epithelium><airway epithelium inflammation><airway inflammation><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-viral agents><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-virals><antiviral compound><antiviral medication><antiviral therapeutic><autosome><biological signal transduction><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><block viral entry><cAMP><cell type><corona virus disease 2019><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 associated death><coronavirus disease 2019 associated fatality><coronavirus disease 2019 associated mortality><coronavirus disease 2019 death><coronavirus disease 2019 disease severity><coronavirus disease 2019 fatality><coronavirus disease 2019 induced death><coronavirus disease 2019 induced fatality><coronavirus disease 2019 induced mortality><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 mortality><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 related death><coronavirus disease 2019 related fatality><coronavirus disease 2019 related mortality><coronavirus disease 2019 severity><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 virus><coronavirus disease infected patient><coronavirus disease patient><coronavirus disease positive patient><coronavirus disease severity><coronavirus disease-19><coronavirus disease-19 mortality><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus infectious disease-19><coronavirus patient><cystic fibrosis transmembrane regulator><cytokine release syndrome><cytokine storm><death due to COVID-19><death due to COVID19><death due to SARS-CoV-2><death due to coronavirus disease 2019><death due to severe acute respiratory syndrome coronavirus 2><death in COVID><death in COVID-19><death in SARS-CoV-2><death in coronavirus disease><death in coronavirus disease 2019><death in severe acute respiratory syndrome coronavirus 2><developmental><driving><drug/agent><epithelial Na+ channel><epithelial sodium channel><fatality due to COVID-19><fatality due to COVID19><fatality due to SARS-CoV-2><fatality due to coronavirus disease 2019><fatality due to severe acute respiratory syndrome coronavirus 2><fluid><gene function><genetic condition><genetic disorder><genetic recessive><genome mutation><global health><hCoV19><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><inhibit viral entry><life-threatening COVID><life-threatening COVID-19><life-threatening SARS-CoV-2><life-threatening coronavirus disease><life-threatening coronavirus disease 2019><life-threatening severe acute respiratory syndrome coronavirus 2><liquid><model of animal><mortality due to COVID-19><mortality due to COVID19><mortality due to SARS-CoV-2><mortality due to coronavirus disease 2019><mortality due to severe acute respiratory syndrome coronavirus 2><nCoV2><nano-molar><nanomolar><novel><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><plasmalemma><prevent><preventing><protein 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mortality><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 mortality><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><severe acute respiratory syndrome coronavirus 2 related death><severe acute respiratory syndrome coronavirus 2 related fatality><severe acute respiratory syndrome coronavirus 2 related mortality><severe acute respiratory syndrome coronavirus 2 severity><severe coronavirus disease><severe coronavirus disease 19><severe coronavirus disease 2019><severe severe acute respiratory syndrome coronavirus 2><stem cells><tool><trans acting element><viral entry blocker><viral entry inhibitor>