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Principal Investigator: Daniel F Dwyer
Organization: BRIGHAM AND WOMEN'S HOSPITAL
Fiscal Year: 2024
Award: $715,968
Funding agency: National Heart Lung and Blood Institute
Project Summary/Abstract:
Mast cells (MCs) expand within the airway epithelium and sub-epithelium during prevalent and burdensome
human respiratory disease, including asthma and nasal polyposis, where they are thought to play a central role
in disease pathobiology. MCs take on discrete protease expression profiles in each location, with sup-epithelial
MCs co-expressing tryptase and chymase (MCTC) while epithelial MCs express tryptase alone (MCT). Neither
the mechanisms underlying this expansion, the signals directing the epithelial and sub-epithelial MC
phenotypes, nor the differential contribution of MC phenotypes to tissue inflammation are well understood. This
application focuses on understanding the mechanisms through which airway tissue structural cells differentially
direct the expansion and differentiation of MC progenitors (MCps), based on preliminary studies identifying a
central role for fibroblasts and epithelial cells in both processes. This proposal tests the central hypothesis that
MC hyperplasia in inflamed human airway mucosa is driven by the recruitment of MCps exhibiting a robust
proliferative capacity, and that the proliferation and differentiation of these recruited MCps towards the
histochemically recognized MCT and MCTC subsets are driven by key signals secreted by airway structural
cells. A related hypothesis is that the resulting intraepithelial and subepithelial MC phenotypes are maintained
through a network of transcription factors regulated by these tissue stromal-derived signals, and that signal
nucleotide polymorphisms in these transcription factors increase risk of developing airway disease. Aim 1 of
this proposal evaluates MCp concentration in human sinonasal tissue across disease endotypes, determining
the relationship between recruited MCp and the proliferative population of MCs we previously identified in
nasal polyposis. Aim 2 of this proposal tests the impact of a series of candidate stromal-derived ligands on
directing MCp proliferation and differentiation using CRISPR/Cas9 gene editing. Aim 3 probes the transcription
factor network underlying MC polarization via shRNA knockdown and evaluates the impact of key asthma-
associated single nucleotide polymorphisms in one such transcription factor on intraepithelial MC
differentiation. Completion of these aims will greatly expand our understanding of the pathways through which
MCs are capable of influencing tissue inflammation and the mechanism(s) through which these pathways are
regulated by their tissue microenvironment.
Terms: <Acetylsalicylic Acid><Address><Airway Disease><Airway mucosa><Anabolism><Aspirin><Asthma><Automobile Driving><B Cell Differentiation Factor I><B cell growth factor 2><B-Cell Growth Factor-II><BCGF-II><BCGF2><Basal Transcription Factor><Basal transcription factor genes><Body Tissues><Bone-Derived Transforming Growth Factor><Bronchial Asthma><CD34><CD34 gene><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><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Differentiation><Cell Differentiation process><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cell Signaling><Cell surface><Cells><Cellular Proliferation><Characteristics><Chemotactic Cytokines><Chymase><Circulation><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><Co-culture><Cocultivation><Coculture><Coculture Techniques><Connective Tissue><Cys-LT><Development><Disease><Disease Progression><Disorder><Effector Cell><Eicosanoids><Eo-CSF><Eosinophil Differentiation Factor><Eosinophilic Esophagitis><Epithelial Cells><Epithelium><Esteroproteases><Exhibits><Family><Fibroblasts><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Generalized Growth><Genes><Genetic Polymorphism><Genetic Transcription><Growth><HPCA1><Histamine><Homologous Chemotactic Cytokines><Human><IL-13><IL-5><IL13><IgA enhancing factor><In Situ><In Vitro><Infiltration><Inflammation><Inflammatory><Intercrines><Interleukin 5 Precursor><Interleukin-13><Interleukin-5><Intracellular Communication and Signaling><JV15-2><Ligands><Link><Location><Lung><Lung Respiratory System><MADH3><MADH3 gene><MMCP-1><Marrow Mast Cell><Mast-Cell Disease><Mediating><Mice><Mice Mammals><Milk Growth Factor><Modern Man><Mothers Against Decapentaplegic, Drosophila, Homolog of, 3><Mucosa><Mucosal Tissue><Mucous Membrane><Multiple Polyps><Murine><Mus><NSAIDs><Nasal><Nasal Cavity Polyp><Nasal Passages Nose><Nasal Polyps><Non-Steroidal Anti-Inflammatory Agents><Nose><Nucleotides><PBMC><PGD2><Pathway interactions><Peptidases><Peptide Hydrolases><Peripheral><Peripheral Blood Mononuclear Cell><Phenotype><Platelet Transforming Growth Factor><Play><Polyps><Population><Process><Production><Progenitor Cells><Proliferating><Prostaglandin D2><Protease Gene><Proteases><Proteinases><Proteolytic Enzymes><Publishing><RNA Expression><Reaction><Reporting><Respiration Disorders><Respiratory Disease><Respiratory Disorder><Respiratory Epithelium><Respiratory Mucosa><Respiratory System Disease><Respiratory System Disorder><Respiratory System, Nose, Nasal Passages><Risk><Role><SIS cytokines><SMA- and MAD-Related Protein 3><SMAD3><Series><Severities><Severity of illness><Signal Transduction><Signal Transduction Systems><Signaling><Single Base Polymorphism><Single Nucleotide Polymorphism><Stromal Cells><Structure of respiratory epithelium><System><T cell replacing factor><T-Cell Replacing Factor><TGF B><TGF-beta><TGF-β><TGFbeta><TGFβ><Testing><Therapeutic><Time><Tissue Basophils><Tissue Growth><Tissues><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Transforming Growth Factor beta><Transforming Growth Factor-Beta Family Gene><Tryptase><WNT Signaling Pathway><WNT signaling><airway epithelium><airway epithelium inflammation><airway inflammation><asthmatic><biological signal transduction><biosynthesis><breathing disorder><cellular differentiation><chemoattractant cytokine><chemokine><chronic rhinosinusitis><chymase-1><chymotrypsin-like protease><co-morbid><co-morbidity><comorbidity><cysteinyl-leukotriene><cytokine><developmental><differential expression><differentially expressed><disease severity><driving><experiment><experimental research><experimental study><experiments><falls><human disease><in vivo><intraepithelial><knock-down><knockdown><lung tryptase><mast cell><mast cell hyperplasia><mast cell protease><mast cell protease 1><mast cell protease I><mast cell protease II><mast cell proteinase-1><mast cell tryptase><mastocyte><mastocytosis><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapy approaches><new treatment approach><new treatment strategy><non-steroidal anti-inflammatory drugs><novel><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapy approach><ontogeny><pathway><polarized cell><polymorphism><polyposis><progenitor><progenitor cell differentiation><progenitor cell proliferation><progenitor differentiation><progenitor proliferation><pulmonary><recruit><respiratory><respiratory dysfunction><respiratory inflammation><respiratory tract epithelium><respiratory tract inflammation><shRNA><short hairpin RNA><single nucleotide variant><skeletal muscle protease><skin tryptase><small hairpin RNA><social role><stem and progenitor cell proliferation><stem and progenitor differentiation><stem cell differentiation><stem cell proliferation><stem cells><transcription factor><transcriptional differences><transcriptomics>