Document text
Principal Investigator: Mary Jane Thomassen
Organization: EAST CAROLINA UNIVERSITY
Fiscal Year: 2019
Award: $311,326
Funding agency: National Institute of Environmental Health Sciences
DESCRIPTION (provided by applicant): Pulmonary granuloma formation is a key feature of sarcoidosis, a chronic disease of unknown etiology. Sarcoidosis has been linked to environmental risk factors (wood-burning stoves, fireplaces and firefighting) that favor carbon nanotube formation. We have shown that lungs instilled with multiwall carbon nanotubes (MWCNT) produce chronic inflammation and granuloma formation resembling sarcoidosis. Sarcoidosis alveolar macrophages exhibit elevated pro-inflammatory cytokines such as CCL5 and IFN-γ, but are deficient in PPARγ, a lipid regulator that regulates the ABC transporters A1/G1 and dampens inflammation. This phenotype appears in macrophages from MWCNT-instilled mice. In macrophage-specific PPARγ-KO mice, inflammatory mediators and granulomatous disease are exacerbated, suggesting that PPARγ is protective. Preliminary data indicate elevated microRNA (miR)-33 in macrophages and granulomatous tissues from both sarcoid patients and MWCNT-instilled mice. The role of miR-33 in the lung is unknown, but elsewhere miR-33 is shown to increase inflammation and repress ABCA1 and ABCG1 lipid transporters (which have anti- inflammatory properties). Strikingly, both transporters are depressed in macrophages from sarcoid patients and MWCNT-instilled mice. Further, in vitro miR-33 overexpression in wild type alveolar macrophages decreases ABCA1 and ABCG1 but not PPARγ. These results suggest that two pathways may dysregulate transporters in granulomatous disease - one associated with intrinsic PPARγ status and the other via environmentally triggered miR-33. We hypothesize that granuloma-protective pathways of PPARγ are mediated by anti-inflammatory actions of ABCA1 and/or ABCG1. Aim 1 will determine functional significance of lipid transporters in granulomatous disease. MWCNT will be instilled in macrophage-specific KO models of: PPARγ, ABCA1, ABCG1, and ABCA1/ABCG1 (double KO) plus wild-type mice. Sixty days later, untreated, sham-(vehicle) treated, or MWCNT-instilled mice will be examined for granuloma size/frequency, and alveolar macrophage expression of ABCA1, ABCG1, miR-33, CCL5, IFNγ, PPARγ, and intracellular cholesterol. Aim 2 will explore the role of miR-33 as a mediator in granulomatous disease and alveolar macrophage expression of ABCA1 and ABCG1. MWCNT-instilled wild-type and macrophage-specific PPARγ KO mice will receive lenti-control, lenti-miR-33, or lenti-miR-33 antagomir. Granuloma size/frequency and alveolar macrophage markers will be evaluated. Subaims will examine miR-33 and lipid transporters in murine and healthy human alveolar macrophages cultured with MWCNT. Aim 3 will determine whether ABCA1, ABCG1, and miR-33 are intrinsically dysregulated in sarcoid alveolar macrophages compared to healthy controls. Subaims will examine effects of lenti-ABCA1, lenti-ABCG1, or lenti-miR-33 antagomir on alveolar macrophages. Our murine MWCNT granuloma model provides a unique opportunity for defining mechanisms and components perpetuating chronicity in environmental lung disease.
Terms: <ABC Transport Protein><ABC Transporter Protein><ABC Transporters><ABCA1><ABCA1 protein><ATP binding cassette transporter 1><ATP-Binding Cassette Transporters><Address><Affect><African American><Afro American><Afroamerican><Air><Alveolar Macrophages><Animal Model><Animal Models and Related Studies><Anti-inflammatory><Antiinflammatory Effect><Area><Basal Transcription Factor><Basal transcription factor genes><Besnier-Boeck Disease><Binding><Black Populations><Body Tissues><Boeck's Sarcoid><Bronchioalveolar Lavage><Bronchoalveolar Lavage><Bronchopulmonary Lavage><Buckytubes><CCL5><Carbon><Carbon Nanotubes><Carbon nano tubes><Carbon nanoparticle><Caucasian><Caucasian Race><Caucasians><Caucasoid><Caucasoid Race><Causality><Cell Body><Cells><Chemokine (C-C Motif) Ligand 5><Cholesterol><Chronic><Chronic Disease><Chronic Granulomatous Disease><Chronic Illness><D17S136E><Data><Depressed mood><Development><Diesel Fuels><Disease><Disease Marker><Disease Pathway><Disorder><Dysfunction><Environment><Environmental Factor><Environmental Health><Environmental Health Science><Environmental Risk Factor><Epidemiologic Research><Epidemiologic Studies><Epidemiological Studies><Epidemiology Research><Etiology><Exhibits><Frequencies><Functional disorder><Gamma interferon><General Transcription Factor Gene><General Transcription Factors><Grant><Granuloma><Granulomatous><Granulomatous Lesion><HDLDT1><Human><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><Immune Interferon><In Vitro><Incidence><Inflammation><Inflammation Mediators><Inflammatory><Inflammatory Response><Interferon Gamma><Interferon Type II><Interferon-gamma><KO mice><Knock-out Mice><Knockout Mice><Lentivirinae><Lentivirus><Link><Lipids><Lung><Lung Granuloma><Lung Lavage><Lung Respiratory System><Lung diseases><MGC17164><Mediating><Mediator><Mediator of Activation><Mediator of activation protein><Methane><Mice><Mice Mammals><Micro RNA><MicroRNAs><Modeling><Modern Man><Molecular Interaction><Murine><Mus><Natural Gas><Null Mouse><Occidental><PPAR gamma><PPAR-γ><PPARgamma><PPARγ><Pathway interactions><Patients><Peroxisome Proliferative Activated Receptor Gamma><Peroxisome Proliferator-Activated Receptor gamma><Peroxisome Proliferator-Activated Receptor γ><Phenotype><Physiopathology><Procedures><Prognosis><Propane><Property><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Macrophages><RANTES><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><Role><SCYA5><SIS delta><SIS-delta><SISd><Sarcoidosis><Schaumann's Disease><Small Inducible Cytokine A5><Subfamily lentivirinae><T-Cell RANTES Protein><T-Cell Specific Protein p288><TCP228><Thiazolidinedione Receptor><Time><Tissues><Transcription Factor Proto-Oncogene><Transcription factor genes><Up-Regulation><Upregulation><Virus-Lenti><Wild Type Mouse><Wood burning stove><Wood stove><Woodstove><anti-inflammatory effect><antiinflammatory><black American><bronchopulmonary lavage therapy><causation><cholesterol transporters><cholesterol-efflux regulatory protein><chronic disorder><consumer product><cytokine><depressed><developmental><disease causation><disease of the lung><disorder of the lung><environmental risk><epidemiologic investigation><epidemiology study><exhaust><fat metabolism><inflammatory mediator><lFN-Gamma><lipid metabolism><lung disorder><macrophage><miRNA><miRNAs><model of animal><model organism><mouse model><murine model><nano materials><nanomaterials><novel><outcome forecast><overexpress><overexpression><particle><pathophysiology><pathway><public health relevance><pulmonary><pulmonary granuloma><respiratory><sadness><social role><therapeutic target><transcription factor><vapor><white race>