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Principal Investigator: Ekaterina Koroleva
Organization: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
Fiscal Year: 2023
Award: $190,161
Funding agency: National Institute of Allergy and Infectious Diseases
Lung immunopathology is a major cause of the morbidity and mortality associated with the SARS-CoV-2
infection. Accumulating evidence suggests that SARS-CoV-2-associated lung damage is caused not only
by the virus, but also by excessive production of proinflammatory cytokines, known as cytokine storm.
Although available vaccines and antiviral drugs protect against infection, these strategies do not specifically
target immune-mediated pathology. Therefore, uncoupling anti-viral host defense mechanisms from the
immunopathology induced by these mechanisms, represents a novel therapeutic strategy for COVID-19
treatment. However, to develop such strategies, a better understanding of the fundamental mechanisms that
regulate SARS-CoV-2-induced immunopathology using animal models of COVID-19 disease is critical. The
critical gap is limited therapeutic approaches that specifically target immune-mediated pathology and
availability of animal models that recapitulate human lung disease. Experiments with mouse models
expressing SARS-CoV-2 receptor, human ACE2 (hACE2) demonstrated virus invasion to the brain and
multiple organ pathology with limited lung pathology, which does not fully recapitulate acute respiratory
distress syndrome in patients with severe COVID-19 disease. Our ongoing results demonstrate that
lymphotoxin beta receptor (LTR)-deficient mice are protected from SARS-CoV-2-induced
immunopathology. We also found that LTβR promotes cytokine storm and lung damage in another model
of respiratory disease, influenza infection. The objective of this proposal is to develop novel mouse models
that mimic lung disease of COVID-19 patients and to test the efficacy of LTR inhibitor to block SARS-CoV-
2-induced lung damage. Our central hypothesis is that expression of hACE2 in type II alveolar epithelial
cells is required for SARS-CoV-2 induced lung disease and that LTR antagonist inhibits SARS-
CoV-2 induced lung immunopathology. To test this hypothesis, we propose two specific aims. In Aim 1,
we will generate mice with regulated hACE2 expression in type II alveolar epithelial cells using CRISPR-
Cas9 system. We will infect these mice intranasally with SARS-CoV-2 and evaluate lung immunopathology
and cytokine expression. In Aim 2, we will test the effectiveness of LTR antagonist to block SARS-CoV-
2-induced immunopathology. We will optimize dose and timing of LTR inhibitors and evaluate lung
immunopathology, viral replication, cytokine production, and protective immunity. This proposal is innovative
and significant, as it will generate novel animal models to study SARS-CoV-2 pathogenesis, provide deeper
understanding of the mechanisms regulating virus-induced immunopathology and test the feasibility of
targeting novel immune regulator, LTR, to inhibit SARS-CoV-2 induced immunopathology without limiting
protective immunity.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><ACE2><ARDS><Acute Respiratory Distress><Acute Respiratory Distress Syndrome><Adult ARDS><Adult RDS><Adult Respiratory Distress Syndrome><Age><Animal Model><Animal Models and Related Studies><Antibody Response><Antiviral Agents><Antiviral Drugs><Antivirals><B cell differentiation factor><B cell stimulating factor 2><B-Cell Differentiation Factor><B-Cell Differentiation Factor-2><B-Cell Stimulatory Factor-2><BCDF><BSF-2><BSF2><BSL-3 facility><BSL3 facility><Biochemical><Biological Response Modifiers><Biomodulators><Body Weight Changes><Brain><Brain Nervous System><CCL2><CCL2 gene><CCL20><CCL20 gene><CCL3><CCL3 gene><COVID infected patient><COVID patient><COVID positive patient><COVID-19><COVID-19 infected patient><COVID-19 infection><COVID-19 patient><COVID-19 positive patient><COVID-19 therapy><COVID-19 treatment><COVID-19 virus><COVID-19 virus infection><COVID19><COVID19 infection><COVID19 patient><COVID19 positive patient><COVID19 therapy><COVID19 treatment><COVID19 virus><CRE Recombinase><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CV-19><CV19><CXCL1><CXCL1 gene><CXCL2><CXCL2 gene><Cachectin Receptors><Cas nuclease technology><Cell Body><Cells><Chemokine (C-C motif) Ligand 3><Chemokine, CC Motif, Ligand 2><Chemokine, CC Motif, Ligand 20><Chemokine, CXC Motif, Ligand 2><Chemotactic Cytokines><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><CoV-2><CoV2><Complex><Consensus><Cytokine Receptors><Da Nang Lung><Data><Development><Disease><Disease model><Disorder><Dose><Encephalon><Enterobacteria phage P1 Cre recombinase><Epithelial Cells><Equilibrium><Exodus 1><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><G0S19-1><GRO Protein, Beta><GRO1><GRO2><GRO2 Oncogene><GROA><GROb><GROβ><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Generations><Genetic><Goals><H1N1><H1N1 Virus><HPGF><Hepatocyte-Stimulating Factor><Histology><Homologous Chemotactic Cytokines><Host Defense Mechanism><Human><Hybridoma Growth Factor><IFN-beta 2><IFNB2><IL-6><IL6 Protein><Immune><Immune Cell Activation><Immune Mediators><Immune Mediators/Modulators><Immune Regulators><Immune Targeting><Immune response><Immunes><Immunity><Immunohistochemistry><Immunohistochemistry Cell/Tissue><Immunohistochemistry Staining Method><Immunological response><Infection><Inflammation><Inflammatory><Influenza A Virus, H1N1 Subtype><Influenza Virus><Intercrines><Interleukin-6><Invaded><KI mice><Knock-in Mouse><LARC><LD78ALPHA><Lung><Lung Alveolar Epithelia><Lung Respiratory System><Lung damage><Lung diseases><MCAF><MCP-1><MCP1><MGI-2><MGSA><MIP 1alpha><MIP-1-alpha><MIP-1a><MIP-2A><MIP1A><MIP2-alpha><MIP2A><MIP2α><MIP3A><Macrophage Inflammatory Protein 3-Alpha><Mediating><Mice><Mice Mammals><Modeling><Modern Man><Monocyte Chemoattractant Protein-1><Monocyte Chemotactic Protein-1><Monocyte Chemotactic and Activating Factor><Monocyte Chemotactic and Activating Protein><Monocyte Chemotactive and Activating Factor><Monocyte Secretory Protein JE><Morbidity><Morbidity - disease rate><Mouse Strains><Murine><Mus><Myeloid Differentiation-Inducing Protein><Organ><Pathologic><Pathology><Pathway interactions><Patients><Plasmacytoma Growth Factor><Production><Proteins><Protocol><Protocols documentation><Public Health><Publishing><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Pathology><RNA Seq><RNA sequencing><RNAseq><Receptor Protein><Receptor Signaling><Reproducibility><Research><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><Role><SARS><SARS corona virus 2><SARS coronavirus disease><SARS-CO-V2><SARS-COVID-2><SARS-CoV disease><SARS-CoV-2><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 inhibitor><SARS-CoV-2 pathogenesis><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 therapy><SARS-CoV-2 treatment><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SCYA2><SCYA20><SCYA3><SCYB1><SCYB2><SIS cytokines><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><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome coronavirus disease><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 infection><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome related corona virus 2><Shock Lung><Small Inducible Cytokine A2><Small Inducible Cytokine A3><Small Inducible Cytokine Subfamily A, Member 20><Small Inducible Cytokine Subfamily B, Member 2><Stem Cell Inhibitor><Stiff lung><System><TNF Receptor Family Protein><TNF Receptor Superfamily><TNF Receptors><TNFR><Testing><Therapeutic><Therapeutic Uses><Transcript Expression Analyses><Transcript Expression Analysis><Transgenic Mice><Tumor Necrosis Factor Receptor><Tumor Necrosis Factor Receptor Family><Tumor Necrosis Factor Receptor Superfamily><Viral><Virus><Virus Replication><Weight Change><Wuhan coronavirus><access to vaccination><access to vaccines><after COVID-19 infection><after SARS-CoV-2 infection><after SARS-CoV2 infection><after infection by SARS-CoV-2><after severe acute respiratory distress syndrome CoV-2 infection><ages><alveolar epithelium><analyze gene expression><angiotensin converting enzyme 2><angiotensin converting enzyme II><antagonism><antagonist><anti-viral agents><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-virals><antiviral compound><antiviral medication><antiviral therapeutic><bacteriophage P1 recombinase Cre><balance><balance function><biosafety level 3 facility><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><chemoattractant cytokine><chemokine><corona virus disease 2019><coronavirus disease 2019><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 therapy><coronavirus disease 2019 treatment><coronavirus disease 2019 virus><coronavirus disease infected patient><coronavirus disease patient><coronavirus disease positive patient><coronavirus disease-19><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus infectious disease-19><coronavirus patient><cytokine><cytokine release syndrome><cytokine storm><design><designing><developmental><disease of the lung><disorder model><disorder of the lung><effective therapy><effective treatment><effectiveness testing><efficacy testing><experiment><experimental research><experimental study><experiments><feasibility testing><flow cytophotometry><flu infection><flu virus infection><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><gene expression analysis><gene expression assay><hCoV19><host response><human tissue><immune activation><immune system response><immunomodulatory biologics><immunopathology><immunoresponse><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 flu><infected with flu virus><infected with influenza><infected with influenza virus><infected with severe acute respiratory syndrome coronavirus 2><influenza infection><influenza virus infection><influenzavirus><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><inhibitor><innovate><innovation><innovative><interferon beta 2><knockin mice><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><lung development><lung disorder><lung injury><lung pathology><lymphotoxin beta receptor><lymphotoxin β receptor><lymphotoxin-beta-specific receptor><macrophage inflammatory protein 2><member><model of animal><mortality><mouse model><murine model><nCoV2><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapy approaches><new treatment approach><new treatment strategy><novel><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapy approach><pathway><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><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><promoter><promotor><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><receptor><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 infected patient><severe acute respiratory syndrome coronavirus 2 pathogenesis><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><severe acute respiratory syndrome coronavirus 2 therapy><severe acute respiratory syndrome coronavirus 2 treatment><severe coronavirus disease><severe coronavirus disease 19><severe coronavirus disease 2019><severe severe acute respiratory syndrome coronavirus 2><sex><social role><therapeutic target><transcriptional profiling><transcriptome sequencing><transcriptomic sequencing><treat COVID-19><treat COVID19><treat SARS-CoV-2><treat coronavirus disease 2019><treat severe acute respiratory syndrome coronavirus 2><vaccination access><vaccination availability><vaccine access><vaccine availability><viral multiplication><viral replication><virus multiplication><wet lung><β-GRO protein>