Document text
Principal Investigator: Sixto Manuel Leal
Organization: UNIVERSITY OF ALABAMA AT BIRMINGHAM
Fiscal Year: 2022
Award: $668,250
Funding agency: National Institute of Allergy and Infectious Diseases
Project summary
Unlike most invasive mold infections, COVID Associated Pulmonary Aspergillosis (CAPA) occurs in
individuals with otherwise intact immune systems suggesting novel biological mechanisms mediating
susceptibility to fungal infection. This proposal seeks to evaluate the novel conceptual advancement that
immune responses targeting intracellular viral pathogens compromise essential human neutrophil
antifungal effector functions required to kill large extracellular fungal hyphae. In Aim 1, we will utilize
BAL samples and autopsy formalin-fixed paraffin embedded (FFPE) lung tissues to characterize the
expression of human neutrophil antifungal effectors, epithelial cell necroptosis, iron content, and fungi in
the respiratory tract during SARS-CoV-2 and CAPA infection. In Aim 2, we will use primary bronchiolar
epithelial cell air-liquid interface (ALI) cultures, CRISPR knockout cell lines (MLKL, RIPK1, RIPK3),
chemical inhibitors of cell death pathways, and neutrophil:fungal killing assays to determine the
mechanism by which SARS-CoV-2 mediates epithelial cell death, iron release, mold growth, and human
neutrophil antifungal effector functions. In Aim 3, we will utilize a novel CAPA infection mouse model,
cre-lox knockdown of human ACE2 expression in neutrophils (MRP8-Cre) and lung epithelial cells
(CC10-Cre) along with iron-deficient fungal mutant strains (ΔsidA, ΔftrA), to evaluate the in vivo impact of
SARS-CoV-2 infection on lung epithelial cell necroptosis, iron release, siderophore-dependent fungal
growth, and neutrophil recruitment/activation. We believe that the results of the proposed study will shed
new light on the fundamental biology mediating viral-associated secondary mold infection which may
enable the development of improved therapeutic regimens that mitigate the risk of developing CAPA.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV variant><2019-nCoV variant forms><2019-nCoV variant strains><A fumigatus><A. fumigatus><ACE2><Air><Allergic Bronchopulmonary Aspergillosis><Anabolism><Antifungal Agents><Antifungal Drug><Aspergillus><Aspergillus fumigatus><Assay><Autopsy><Bioassay><Biologic Assays><Biological><Biological Assay><Biology><Blood Neutrophil><Blood Polymorphonuclear Neutrophil><Body Tissues><Bronchioalveolar Lavage><Bronchioles><Bronchiolus><Bronchoalveolar Lavage><Bronchoalveolar Lavage Fluid><Bronchopulmonary Aspergillosis><Bronchopulmonary Lavage><CAGA><CGLA><COVID><COVID-19 infection><COVID-19 variant><COVID-19 variant forms><COVID-19 variant strains><COVID-19 virus><COVID19 infection><COVID19 virus><CRISPR><CRISPR/Cas system><Cell Culture Techniques><Cell Death><Cell Line><CellLine><Chemicals><Chemoattractants><Chemotactic Cytokines><Chemotactic Factors><Chemotaxins><Clustered Regularly Interspaced Short Palindromic Repeats><CoV disease><CoV-2><CoV2><Data><Development><Dysfunction><Enabling Factors><Epithelial Cells><Evaluation><Exhibits><Fe chelation><Fe element><Ferroprotoporphyrin><Filamentous Fungi><Formalin><Functional disorder><Fungus Diseases><Gene Expression><Generalized Growth><Germination><Growth><Heme><Homologous Chemotactic Cytokines><Host Defense><Human><Hyphae><IFN><Immune><Immune response><Immune system><Immunes><Immunity><Immunocompetent><Immunohistochemistry><Immunohistochemistry Cell/Tissue><Immunohistochemistry Staining Method><Immunological response><In Situ><Individual><Infection><Infection prevention><Intensive Care Units><Intercrines><Interferons><Intervention><Intervention Strategies><Iron><Iron Chelation><Knock-out><Knockout><Laboratories><Light><Liquid substance><Lung><Lung Lavage><Lung Parenchyma><Lung Respiratory System><Lung Tissue><MA387><MRP8><Marrow Neutrophil><Mediating><Mice><Mice Mammals><Modeling><Modern Man><Molds><Murine><Mus><Mycoses><Neutrophil Infiltration><Neutrophil Recruitment><Neutrophilic Granulocyte><Neutrophilic Leukocyte><Nutrient><Oxyhemoglobin><Paraffin Embedding><Pathway interactions><Patients><Pharmacology><Photoradiation><Physiopathology><Polymorphonuclear Cell><Polymorphonuclear Leukocytes><Polymorphonuclear Neutrophils><Predisposition><Prevent infection><Production><Proteins><Protoheme><Pulmonary Body System><Pulmonary Organ System><RIP1><RIP3><RIPK1><RIPK1 gene><RIPK3><RIPK3 gene><Receptor-Interacting Protein><Receptor-Interacting Protein 3><Receptor-Interacting Serine/Threonine Kinase 1><Receptor-Interacting Serine/Threonine Protein Kinase 3><Regimen><Reproduction spores><Research Specimen><Respiratory System><Respiratory Tracts><Respiratory tract structure><Risk><Role><S100A8><S100A8 gene><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 infection><SARS-CoV-2 variant><SARS-CoV-2 variant forms><SARS-CoV-2 variant strains><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><SIS cytokines><Sampling><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 corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Siderochromes><Siderophores><Specimen><Spores><Strains Cell Lines><Structure of parenchyma of lung><Susceptibility><Testing><Therapeutic><Therapeutic Fungicides><Tissue Growth><Tissues><Viral><Viral Diseases><Virus Diseases><Wuhan coronavirus><allergic/immunologic body system><allergic/immunologic organ system><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-fungal><anti-fungal agents><anti-fungal drug><antifungals><biologic><biosynthesis><bronchial epithelium><bronchopulmonary lavage therapy><cell culture><cell cultures><chemoattractant cytokine><chemokine><complement chemotactic factor><corona virus disease><coronavirus disease><coronavirus disease 2019 infection><coronavirus disease 2019 variant><coronavirus disease 2019 variant forms><coronavirus disease 2019 variant strains><coronavirus disease 2019 virus><coronavirus disease-19 virus><cultured cell line><cytokine><developmental><digital><extracellular><ferroheme><fluid><fungal infection><fungal pathogen><fungi pathogen><fungus><fungus infection><hCoV19><host response><immune competent><immune system response><immunoresponse><improved><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><inhibitor><innovate><innovation><innovative><interventional strategy><knock-down><knockdown><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><mortality><mouse model><murine model><mutant><nCoV2><nano-string><nanostring><necrocytosis><necropsy><neutrophil><novel><ontogeny><pathogenic fungus><pathogenic virus><pathophysiology><pathway><postmortem><prevent><preventing><pulmonary><pulmonary aspergillosis><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 variant><severe acute respiratory syndrome coronavirus 2 variant forms><severe acute respiratory syndrome coronavirus 2 variant strains><severe coronavirus disease><severe coronavirus disease 19><severe coronavirus disease 2019><severe severe acute respiratory syndrome coronavirus 2><social role><transcriptomics><viral infection><viral pathogen><virus infection><virus pathogen><virus-induced disease>