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Principal Investigator: Emanuela Marina Bruscia
Organization: YALE UNIVERSITY
Fiscal Year: 2020
Award: $412,285
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
Macrophages (MΦs) kill microorganisms, engulf dead cells and debris, and regulate the immune response.
They are thus gatekeepers of tissue health, including the lungs. The lung-tissue-resident MΦs (TR-MΦs) are
the interstitial and alveolar MΦs, which have complementary but distinct functions. In response to infections,
lungs are rapidly populated by waves of Ly6C+ circulating monocytes. In concert with TR-MΦs, these
monocytes fight the infection, then facilitate resolution of the inflammatory response. Many chronic lung
inflammatory diseases, including cystic fibrosis (CF), are associated with dysregulated MΦ function. Our long-
term goal is to understand how different lung MΦ populations contribute to lung hyper-inflammation and
infection, and to elucidate the biology of these distinct cell populations. The objective of this proposal is to
characterize ezrin’s role in monocyte/MΦ function. Our central hypothesis is that ezrin controls monocyte/MΦ
cortical actin organization and signal transduction events in response to inflammatory/infectious stimuli. These
cellular changes allow the MΦs to spread, move, phagocytize, and survive, thus shaping the magnitude and
quality of the lung immune response to infections. The rationale for these studies is that low ezrin levels have
been found in MΦs from patients with CF (our own work). Other investigators have also reported low ezrin
levels in blood cells from individuals with asthma. Thus, by elucidating the molecular mechanism by which ezrin
shapes lung MΦ functions, we could identify potential therapeutic targets for lung diseases. Our specific aims
will test the following hypotheses: (Aim 1) ezrin is necessary for monocyte/MΦ adaptation to the inflamed lung
microenvironment; (Aim 2) ezrin is needed for efficient phagocytosis of Staphylococcus aureus and
Pseudomonas aeruginosa, two microorganisms that CF patients fail to efficiently eradicate from their lungs;
(Aim 3) functional CFTR, the gene that causes CF when mutated, is needed to preserve normal ezrin levels
during MΦ activation. The contribution is significant since very little is known about ezrin’s role in regulating
lung MΦ activation. Our proposed research is innovative because we will use an unprecedented mouse model
in which ezrin is knocked out specifically in monocytes and MΦs. Thus, the proposed studies will investigate in
depth the consequences of ezrin loss in monocytes and MΦs during lung infection and inflammation.
Terms: <3D cell culture><3D culture><Actin-Binding Protein><Actins><Adhesions><Affect><Alveolar><Apoptosis><Apoptosis Pathway><Asthma><Bacteria><Binding><Biology><Blood Cells><Blood monocyte><Body Tissues><Bronchial Asthma><CF lung disease><CF patients><CFTR><CFTR Protein><Cause of Death><Cell Body><Cell Communication and Signaling><Cell Function><Cell Process><Cell Shape><Cell Signaling><Cell membrane><Cell physiology><Cell-Extracellular Matrix><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chlorides><Chronic><Clinical><Complex><Cystic Fibrosis><Cystic Fibrosis Transmembrane Conductance Regulator><Cytoplasmic Membrane><Data><Disease><Disorder><ECM><Event><Extracellular Matrix><F-Actin><Filamentous Actin><Filopodia><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Gatekeeping><Gene Alteration><Gene Mutation><Genetic Alteration><Genetic Change><Genetic defect><Goals><Health><Homolog of Drosophila TOLL><Human><Image><Immune response><Immunological response><Immunomodulation><Impairment><In Vitro><Individual><Infection><Inflammation><Inflammatory><Inflammatory Response><Inhalation><Inhaling><Inositide Phospholipids><Inositol Phosphoglycerides><Inositol Phospholipids><Intracellular Communication and Signaling><Investigation><Investigators><KO mice><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Laboratories><Lipopolysaccharides><Lung><Lung Inflammation><Lung Parenchyma><Lung Respiratory System><Lung Tissue><Lung diseases><Lung infections><Macromolecular Complexes><Marrow monocyte><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Metabolic Protein Degradation><Mice><Mice Mammals><Microscopy><Modern Man><Molecular><Molecular Interaction><Morphologic Finding><Mucoviscidosis><Murine><Mus><Mutate><Mutation><Nebulizer><Null Mouse><P aeruginosa><P. aeruginosa><PI-3K/AKT><PI3K/AKT><PIP2><Peripheral Blood Cell><Phagocytes><Phagocytic Cell><Phagocytosis><Phosphatidyl Inositol><Phosphatidylinositol 4,5-Biphosphate><Phosphatidylinositol 4,5-Diphosphate><Phosphatidylinositol-4,5-Bisphosphate><Phosphatidylinositols><Phosphoinositides><Phosphorylation><Physical shape><Plasma Membrane><Population><Production><Programmed Cell Death><Protein Family><Protein Phosphorylation><Protein Turnover><Proteins><Pseudomonas aeruginosa><Pseudomonas pyocyanea><PtIns 4,5-P2><PtdIns><PtdInsP2><Publishing><Pulmonary Cystic Fibrosis><Pulmonary Diseases><Pulmonary Disorder><Regulation><Regulator Genes><Regulatory Protein Degradation><Reporting><Research><Research Personnel><Researchers><Resolution><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><Role><S aureus><S. aureus><Shapes><Signal Transduction><Signal Transduction Systems><Signaling><Spatial Distribution><Staph aureus><Staphylococcus aureus><Stimulus><Structure><Structure of parenchyma of lung><Subcellular Process><Surface Proteins><TLR4><TLR4 gene><Techniques><Testing><Tissues><Toll Homologue><Transcriptional Regulatory Elements><Work><amebocyte><bactericidal><bactericide><base><biological signal transduction><chemical property><cystic fibrosis lung disease><cystic fibrosis patients><cystic fibrosis transmembrane regulator><cytokine><disease of the lung><disorder of the lung><ezrin><fighting><flow cytophotometry><gatekeeper><genetic approach><genetic strategy><genome mutation><host response><imaging><immune modulation><immune regulation><immune regulator><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><in vitro testing><in vivo><in vivo evaluation><in vivo testing><individuals with CF><individuals with cystic fibrosis><inflammatory lung disease><innovate><innovation><innovative><interstitial><lung disorder><macrophage><member><membrane-organizing extension spike protein><microorganism><migration><moesin><monocyte><morphologic criteria><morphologic signature><morphological criteria><morphological signature><mouse model><murine model><patients with CF><patients with cystic fibrosis><phosphoprotein p81><plasmalemma><preservation><protein degradation><pulmonary><pulmonary infections><radixin><radixin protein><regulatory gene><response><small molecule><social role><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic target><three dimensional cell culture><toll-like receptor 4><trafficking><trans acting element>