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Principal Investigator: Allison Marie Greaney
Organization: YALE UNIVERSITY
Fiscal Year: 2024
Award: $76,756
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY/ABSTRACT
Fibrotic lung remodeling is a hallmark of many lung diseases, including severe influenza infection, idiopathic
pulmonary fibrosis, and COVID-19. While a basal-like epithelial cell has been identified as a central player to this
aberrant repair response, little has been done to investigate the behavior and interactions of other cells in the
diseased tissue, particularly alveolar macrophages. Therefore, the goal of this study is to leverage a biomimetic
engineered lung model system to investigate the relationship between regenerating basal-like progenitor cells
and pulmonary macrophages. This engineered lung tissue system is based on cellular repopulation and culture
of a decellularized native rat lung scaffold. This platform enables a well-controlled, native-like tissue environment
for the evaluation of cell-cell interactions, without the systemic confounders of in vivo studies. It is expected that
macrophages will significantly influence and direct epithelial remodeling by these basal-like cells, particularly in
relation to the fibrotic or anti-fibrotic activation state of the macrophages. This work expands on previous findings
that adding macrophages to engineered lung cultures containing basal-like progenitor cells significantly improves
tissue architecture and regenerative epithelial cell phenotype, compared to engineered lung cultures without
macrophages. First, pulmonary macrophages will be isolated from rats by bronchoavleolar lavage and
characterized. Protocols will be developed to chemically stimulate macrophages in vitro to a disease-like
inflammatory state, or to a reparative anti-fibrotic state. Next, macrophages of different activation states will be
introduced to air-liquid interface cultures of regenerative basal cells, to evaluate epithelial-macrophage
interactions in isolation. Finally, activated macrophages will be introduced to engineered lung cultures containing
regenerative basal cells, fibroblasts, and endothelium to recapitulate essential native cellular communities.
Engineered lung tissues will be evaluated for histologic and biomechanic changes between conditions, as well
as differential cell signaling patterns, as evaluated by single-cell RNA sequencing. It is expected that
inflammatory macrophages will contribute to fibrotic response in regenerating epithelium, whereas anti-fibrotic
macrophages will contribute to more functional alveolar regeneration by basal-like cells. The findings of this
study will elucidate the role of pulmonary macrophages in governing lung repair and regeneration in this model
system. Further, this work may suggest possible routes for the treatment of fibrotic lung diseases. The proposed
research project will be executed by Allison M. Greaney at the David H. Koch Institute for Integrative Cancer
Research at the Massachusetts Institute of Technology (MIT), under the Sponsorship of Dr. Robert Langer, and
Co-Sponsorship of Dr. Ruslan Medzhitov at Yale University. Dr. Langer and Dr. Medzhitov will mentor Allison in
her Research and Professional Training Goals to develop new research and scientific communication skills, so
she may be well-equipped to secure a tenure-track faculty position at a top research institution.
Terms: <Air><Alveolar><Alveolar Cell><Alveolar Macrophages><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Apoptotic><Architecture><Autoregulation><B cell growth factor><B-Cell Differentiation Factor-1><B-Cell Growth Factor-1><B-Cell Growth Factor-I><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><BCSF 1><BSF-1><BSF1><Basal Cell><Behavior><Benchmarking><Best Practice Analysis><Binetrakin><Biologic Models><Biological Mimetics><Biological Models><Biomechanics><Biomedical Engineering><Biomimetics><Body Tissues><Bronchioalveolar Lavage><Bronchoalveolar Lavage><Bronchopulmonary Lavage><COVID-19><COVID-19 infection><COVID-19 virus infection><COVID19 infection><CV-19><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Interaction><Cell Signaling><Cell-to-Cell Interaction><Cells><Cessation of life><Chemical Stimulation><Chemicals><Co-culture><Cocultivation><Coculture><Coculture Techniques><Common Rat Strains><Communication><Coronavirus Infectious Disease 2019><Data><Death><Dinoprostone><Disease><Disease Outcome><Disorder><Distal><Embryonic Muscle Cells><Endothelium><Engineering><Engineering / Architecture><Environment><Epithelial Cells><Epithelium><Evaluation><Faculty><Fibroblasts><Fibrosing Alveolitis><Fibrosis><Fibrotic lesions in lung><Goals><Histologic><Histologically><Homeostasis><Human><Hypoxemia><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><IL-4><IL4 Protein><Immune Interferon><In Vitro><Inflammatory><Injury><Institution><Interferon Gamma><Interferon Type II><Interleukin-4><Interleukin-4 Precursor><Intervention><Intervention Strategies><Intracellular Communication and Signaling><Lavage><Lipopolysaccharides><Liquid substance><Lung><Lung Alveolar Epithelia><Lung Diseases><Lung Lavage><Lung Parenchyma><Lung Respiratory System><Lung Tissue><Lung scar><Lung tissue regeneration><Lung tissue scar><Lymphocyte Stimulatory Factor 1><MCGF-2><Macrophage><Maintenance><Massachusetts><Mast Cell Growth Factor-2><Mentors><Mesenchymal><Mesenchymas><Mesenchyme><Methods><Model System><Modern Man><Myoblasts><Myofibroblast><Mφ><Natural regeneration><PGE2><PGE2 alpha><PGE2alpha><Paracrine Communication><Paracrine Signaling><Pattern><Phenotype><Physiological Homeostasis><Play><Population><Position><Positioning Attribute><Precursor Muscle Cells><Production><Progenitor Cells><Prostaglandin E2><Prostaglandin E2 alpha><Prostaglandin E2alpha><Proteins><Protocol><Protocols documentation><Publishing><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Macrophages><Pulmonary Scar><Pulmonary Tissue fibrosis><R-Series Research Projects><R01 Mechanism><R01 Program><Rat><Rats Mammals><Rattus><Regeneration><Reporting><Research><Research Grants><Research Project Grants><Research Projects><Role><Route><SARS-CoV-2 infection><SARS-CoV2 infection><Scarring at the lung><Scarring in the lung><Secure><Severe acute respiratory syndrome coronavirus 2 infection><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skeletal muscle injury><Structure of parenchyma of lung><System><T-Cell Growth Factor 2><Technology><Therapeutic Intervention><Tissue Model><Tissues><Training><Universities><Vascular Endothelium><Work><alveolar epithelium><anti-cancer research><benchmark><bio-engineered><bio-engineers><bioengineering><biological engineering><biological signal transduction><biomechanical><bronchopulmonary lavage therapy><cancer research><cell community><cell type><cellular community><coronavirus disease 2019><coronavirus disease 2019 infection><coronavirus disease-19><coronavirus infectious disease-19><critical injury><cytokine><devastating injury><diffuse interstitial pulmonary fibrosis><disease of the lung><disorder of the lung><epithelial progenitor><epithelial progenitor cell><epithelial repair><epithelial stem cell><epithelium regeneration><fibrotic lung><fibrotic lung disease><fibrotic pulmonary disease><flu infection><flu virus infection><fluid><hypoxemic><idiopathic pulmonary fibrosis><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 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><injuries><insight><intervention therapy><interventional strategy><lFN-Gamma><lavage therapy><liquid><lung disorder><lung regeneration><lung repair><lung tissue repair><migration><muscle fiber repair><muscle repair><muscle tissue repair><muscular repair><novel><organ development><organ growth><pharmacologic><progenitor><pulmonary><pulmonary regeneration><pulmonary repair><recruit><regenerate><regenerate epithelium><regenerative><repair><repaired><response><restoration><scRNA-seq><scaffold><scaffolding><severe injury><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><skills><social role><stem cells><tenure process><tenure track>