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Principal Investigator: Steven Michael Zwick
Organization: MASSACHUSETTS GENERAL HOSPITAL
Fiscal Year: 2024
Award: $76,984
Funding agency: National Heart Lung and Blood Institute
A significant fraction of asthma patients face challenges in managing their symptoms with available therapies.
Uncovering new drug targets holds the promise of alleviating their suffering and improving their quality of life.
Anticholinergics, which bind and inhibit muscarinic acetylcholine receptors, have shown recent success in
clinical trials. Animal research suggests these drugs may mitigate the lung inflammation and remodeling
observed in asthma, although the underlying mechanisms are unclear. These drugs bind M1 and M3
muscarinic receptor subtypes in the airway epithelium, where each receptor’s role in asthmatic allergen
uptake, inflammation, and remodeling is unknown. We identified secretory cell associated antigen passages
(SAPs) in the airway epithelium, which uptake asthmatic allergens into the epithelium and directly associate
with underlying antigen-presenting cells. Acetylcholine signaling increases the fraction of airway epithelial cells
internalizing allergens into SAPs, but it is not known whether M1 or M3 receptors mediate this response. We
hypothesize that M1 and M3 differentially regulate SAP formation to modulate antigen provision. In mouse
models of asthma, anticholinergic drugs that inhibit M1 and M3 reduce type 2 inflammation and goblet cell
metaplasia, hallmark pathologies of the disease. However, genetic deletion of the M1 or M3 receptor causes
only the reduced inflammatory or metaplastic phenotype, respectively. We therefore hypothesize that epithelial
acetylcholine signaling through the M1 and M3 receptors separately regulates type 2 inflammation and goblet
cell metaplasia during the asthmatic response. To test our hypotheses, we will use mouse genetics to delete
M1 or M3 receptors. By using confocal and two-photon microscopy to visualize fluorescently labeled allergens
in airway epithelial cultures and tracheal explants, we will determine which receptor mediates the stimulation of
SAP formation by acetylcholine. Further, we will generate conditional knockout mice with established
promoters to excise the M1 and M3 receptors specifically in airway epithelial cell types. We will determine the
consequences for allergic response using the house dust mite (HDM)-induced asthma mouse model. We will
measure the expression of type 2 inflammatory markers using flow cytometry, qPCR, and ELISA and will detect
goblet cell metaplasia by immunofluorescence imaging. The training plan outlined will enable me to acquire
advanced skills in murine genetics, airway epithelial culture and physiology, two-photon microscopy, and
immunology. It will also further enhance my abilities in scientific communication. Obtaining these skills is crucial
for my development into an interdisciplinary scientist employing quantitative and biomedical techniques to
study lung disease. The Rajagopal laboratory and the greater MGH community provide an inimitable
environment in which to learn these technical skills and to receive the professional training I need to become
an independent investigator. Dr. Rajagopal’s scientific output, funding, and training of academic scientists
demonstrate a successful track record of mentorship.
Terms: <2-photon microscopy><ACh Receptors><Acetylcholine><Acetylcholine Agents><Acetylcholine Receptors><Acute><Address><Adrenal Cortex Hormones><Affect><Affinity><Agonist><Air Movements><Allergens><Allergy><Animal Experimental Use><Animal Experimentation><Animal Research><Anti-Cholinergics><Anticholinergic Agents><Anticholinergics><Antigen Presentation><Antigen-Presenting Cells><Antigens><Area><Asthma><B Cell Differentiation Factor I><B cell growth factor><B cell growth factor 2><B-Cell Differentiation Factor-1><B-Cell Growth Factor-1><B-Cell Growth Factor-I><B-Cell Growth Factor-II><B-Cell Proliferating Factor><B-Cell Stimulating Factor><B-Cell Stimulating Factor-1><B-Cell Stimulation Factor-1><B-Cell Stimulatory Factor-1><BCDF-1><BCGF><BCGF-1><BCGF-II><BCGF2><BCSF 1><BSF-1><BSF1><Binding><Binetrakin><Biomedical Engineering><Blood Eosinophil><Bronchial Asthma><Cell Body><Cell Communication and Signaling><Cell Isolation><Cell Segregation><Cell Separation><Cell Separation Technology><Cell Signaling><Cells><Characteristics><Cholinergic Agents><Cholinergic Drugs><Cholinergic Receptors><Cholinoceptive Sites><Cholinoceptors><Clinical><Clinical Trials><Communication><Communities><Complication><Corticoids><Corticosteroids><Curiosities><Data><Dermatophagoides Allergens><Development><Disease><Disorder><Drug usage><Drugs><ELISA><Environment><Enzyme-Linked Immunosorbent Assay><Eo-CSF><Eosinophil Differentiation Factor><Eosinophilia><Eosinophilic Granulocyte><Eosinophilic Leukocyte><Epithelial Cells><Epithelium><Face><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Funding><Genetic><Goals><Goblet Cells><HP40><Homolog of Mouse T Cell and Mast Cell Growth Factor 40><House Dust Mite Allergens><House Dust Mites><Housedust Mites><Hypersensitivity><IL-13><IL-4><IL-5><IL-9><IL13><IL4 Protein><IL9 Protein><IgA enhancing factor><Image><Immune><Immunes><Immunofluorescence><Immunofluorescence Immunologic><Immunology><Individual><Inflammation><Inflammation Mediators><Inflammatory><Inflammatory Response><Inhalation><Inhaling><Interleukin 5 Precursor><Interleukin 9 Precursor><Interleukin-13><Interleukin-4><Interleukin-4 Precursor><Interleukin-5><Interleukin-9><Intestinal><Intestines><Intracellular Communication and Signaling><Investigators><Involuntary Muscle><KO mice><Kinetics><Knock-out Mice><Knockout Mice><Label><Laboratories><Learning><Lung><Lung Diseases><Lung Inflammation><Lung Respiratory System><Lymphocyte Stimulatory Factor 1><M1 receptor><M3 receptor><MCGF-2><Marrow Eosinophil><Mast Cell Growth Factor-2><Measures><Mediating><Medication><Mentorship><Metaplasia><Metaplastic Change><Mice><Mice Mammals><Molecular><Molecular Interaction><Molecular Target><Mucous body substance><Mucus><Murine><Mus><Muscarinic Acetylcholine Receptor><Muscarinic M1 Receptor><Muscarinic M3 Receptor><Muscarinic Receptors><Null Mouse><Oral Ingestion><Output><Ovalbumin><Pathology><Persons><Pharmaceutical Preparations><Phenotype><Physiology><Pneumonitis><Public Health><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Inflammation><Pyroglyphidae><QOL><Quality of life><Receptor Protein><Reporting><Research Personnel><Research Proposals><Researchers><Respiratory Epithelium><Role><Scientist><Secretory Cell><Secretory Rate><Signal Transduction><Signal Transduction Systems><Signaling><Site><Smooth Muscle><Structure><Structure of respiratory epithelium><Surface><Symptoms><T cell replacing factor><T-Cell Growth Factor 2><T-Cell Growth Factor P40><T-Cell Replacing Factor><T-Cell/Mast Cell Growth Factor p40><Technical Expertise><Techniques><Testing><Th-2 Cell><Th2 Cells><Therapeutic><Therapeutic Uses><Trachea><Trachea Proper><Training><Type 2 Helper Cell><Visualization><Work><accessory cell><air flow><airflow><airway epithelium><airway smooth muscle><allergen response><allergic response><allergy response><animal experimentations><antagonism><antagonist><asthma model><asthma patient><asthmatic><asthmatic airway><asthmatic patient><bio-engineered><bio-engineers><bioengineering><biological engineering><biological signal transduction><bowel><cell sorting><cell type><chelation><cholinergic><chronic airway disease><chronic respiratory disease><clinical relevance><clinically relevant><conditional knock-out><conditional knockout><constriction><cytokine><developmental><disease of the lung><disorder of the lung><drug use><drug/agent><dust mite allergens><enzyme linked immunoassay><eosinophil><extracellular><faces><facial><flow cytophotometry><imaging><immunogen><improved><improved outcome><inflammation marker><inflammatory marker><inflammatory mediator><lung disorder><mouse genetics><mouse model><mucous><murine model><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><p40 Cytokine><p40 Protein><promoter><promotor><pulmonary><receptor><receptor function><respiratory smooth muscle><respiratory tract epithelium><response><side effect><skills><social role><standard care><standard treatment><success><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><technical skills><two photon excitation microscopy><two photon microscopy><uptake><windpipe>