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Principal Investigator: MARK ROBERTS LOONEY
Organization: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
Fiscal Year: 2023
Award: $623,280
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract
The acute respiratory distress syndrome (ARDS) was first formally described in 1967, but it has likely been the
predominant cause of death in pandemic viral infections for centuries. Now, the world is at grips with a new
pandemic from SARS-CoV-2, or COVID-19, which has infected over 100 million people worldwide resulting in
>2 million deaths. Central to the immunopathogenesis of ARDS is the role of neutrophils and neutrophil
activation, including the release of neutrophil chromatin into the extracellular space in a process termed
neutrophil extracellular traps, or NETs. Originally described as a form of host defense to inactivate pathogens,
NETs have emerged as a potentially maladaptive response to infections, producing substantial bystander injury
to tissues and serving as a nidus for coagulation. We have shown that NETs are produced in response to both
sterile and pathogen-induced acute lung injury (including Influenza A), and when neutralized, lung injury is
reduced without compromising microbial containment. We have also shown that NETs are increased in the
plasma of patients with ARDS and associate with more severe ARDS and ARDS mortality. Emerging reports
indicate that NETs are also present in the blood and lungs of COVID-19 patients. We are now positioned to
rapidly test the role of NETs in response to SARS-CoV-2 in this application, and to develop novel approaches to
neutralize NETs for therapeutic purposes. In Aim 1, we will challenge neutrophils with SARS-CoV-2 virus or
spike protein to determine the production of NETs and NET-induction molecular pathways. We hypothesize that
secreted neutrophil proteases and NETs themselves will cleave spike protein to prime for enhanced viral
pathogenesis. We will also embark on unbiased studies of neutrophil proteins using mass spectrometry to
determine novel pathways of neutrophil activation resulting from SARS-CoV-2, including from neutrophils
isolated from COVID-19 patients. In Aim 2, we will turn our attention to a mouse model of COVID-19 in which
mouse strains expressing human ACE2 will be challenged with SARS-CoV-2 and lung injury, NETs, and
systemic effects measured. We hypothesize that NETs will be produced and predominantly localize to the
alveolar spaces. We will use mice with gain and loss of function mutations in NET pathways (PAD4-/-, DNase1-
/-, DNase1L3-/-) to test for their pathogenicity in this model. In these studies, we will also test a novel DNase to
neutralize NETs into non-toxic mononucleosomes, which could be rapidly deployed to COVID-19 patients.
Finally, we will test for the role of NET-associated platelet activation, complement activation, and coagulation
responses, and therapeutic strategies to mitigate these systemic effects. In summary, these studies will establish
new knowledge on neutrophil activation and the definitive role of NETs in COVID-19 and identify therapeutic
approaches to target NETs in SARS-CoV-2-induced ARDS.
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related acute respiratory distress syndrome><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><COVID19/ARDS><COVID19/acute respiratory distress syndrome><CV-19><CV19><Cause of Death><Cessation of life><Chromatin><Clotting><CoV disease><CoV-2><CoV2><Coagulation><Coagulation Process><Complement><Complement Activation><Complement Proteins><Containment><DNA Nucleases><DNase><Da Nang Lung><Death><Deoxyribonucleases><Development><Disease><Disorder><Esteroproteases><Excess Mortality><Explosion><Extracellular Space><Feedback><Grippe><Grips><Host Defense><Human><Infection><Infection Control><Influenza><Influenza A><Influenza A virus><Influenza Viruses Type A><Influenzavirus A><Intercellular Space><Knowledge><Literature><Lung><Lung Respiratory System><Lung damage><Lung infections><Marrow Neutrophil><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measures><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Mouse Strains><Murine><Mus><Neutrophil Activation><Neutrophilia><Neutrophilic Granulocyte><Neutrophilic Leukocyte><Orthomyxovirus Type A><Pathogenesis><Pathogenicity><Pathway interactions><Patients><Peptidases><Peptide Hydrolases><Persons><Plasma><Plasma Serum><Platelet Activation><Pneumonia><Polymorphonuclear Cell><Polymorphonuclear Leukocytes><Polymorphonuclear Neutrophils><Position><Positioning Attribute><Predisposition><Process><Production><Protease Gene><Proteases><Proteinases><Proteins><Proteolytic Enzymes><Proteomics><Reporting><Reticuloendothelial System, Serum, Plasma><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 associated ARDS><SARS-CoV-2 associated acute respiratory distress syndrome><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 induced ARDS><SARS-CoV-2 induced acute respiratory distress syndrome><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 pandemic><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 related ARDS><SARS-CoV-2 related acute respiratory distress syndrome><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2/ARDS><SARS-CoV-2/acute respiratory distress syndrome><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 epidemic><SARS-CoV2 infection><SARS-CoV2 pandemic><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sepsis><Serum><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 CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><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 epidemic><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Severities><Shock Lung><Sterility><Stiff lung><Susceptibility><Testing><Therapeutic><Trachea><Trachea Proper><Transgenic Organisms><Type A Influenza><Viral Diseases><Viral Pathogenesis><Viral Pneumonia><Virus><Virus Diseases><Wuhan coronavirus><after COVID-19 infection><after SARS-CoV-2 infection><after SARS-CoV2 infection><after infection by SARS-CoV-2><after 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pandemic><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus disease/ARDS><coronavirus disease/acute respiratory distress syndrome><coronavirus infectious disease-19><coronavirus patient><developmental><effective therapy><effective treatment><emergent pandemic><emerging pandemic><experiment><experimental research><experimental study><experiments><extracellular><following COVID-19 infection><following SARS-CoV-2 infection><following SARS-CoV2 infection><following infection by SARS-CoV-2><following severe acute respiratory distress syndrome CoV-2 infection><gain of function><grasp><hCoV19><in vivo><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><injury to tissue><life-threatening COVID><life-threatening COVID-19><life-threatening SARS-CoV-2><life-threatening coronavirus disease><life-threatening coronavirus disease 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disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><peripheral blood><post SARS-CoV-2 infection><prevent><preventing><previous COVID-19 infection><previous SARS-CoV-2 infection><previous SARS-CoV2 infection><previous severe acute respiratory distress syndrome CoV-2 infection><prior COVID-19 infection><prior SARS-CoV-2 infection><prior SARS-CoV2 infection><prior severe acute respiratory distress syndrome CoV-2 infection><pulmonary><pulmonary damage><pulmonary infections><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><respiratory pathogen><response><serious COVID><serious COVID-19><serious SARS-CoV-2><serious coronavirus disease><serious coronavirus disease 2019><serious severe acute respiratory syndrome coronavirus 2><severe COVID><severe COVID-19><severe COVID19><severe SARS-CoV-2><severe acute respiratory syndrome coronavirus 2 associated ARDS><severe acute respiratory syndrome coronavirus 2 associated acute respiratory distress 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target><therapeutic testing><tissue injury><transgenic><viral infection><viral pandemic><virus infection><virus pathogenesis><virus-induced disease><wet lung><windpipe>