Role Of Adenosine In Allergic Lung Disease
Stephen Lloyd Tilley
Medicineuniversity Of North Carolina Chapel Hill
Grant 2R01HL071802-05A1 from National Heart, Lung, And Blood Institute, IRG: LCMI
Abstract: Elevated adenosine levels in the lungs and exhaled breath of asthmatics, with further increases following antigen challenge, suggests that this ubiquitous mediator may contribute to the pathophysiology of asthma. Many of the effects of adenosine in the asthmatic lung are mast cell-dependent. Adenosine has both pro- and anti-inflammatory effects on mast cells, due to the expression of multiple adenosine receptors on the cell surface, each capable of activating very different intracellular signaling pathways. In this proposal we will test the hypothesis that both pro- and anti-inflammatory signals are transmitted to the mast cell by adenosine via the activation of distinct cell-surface receptors, and that engagement of these receptors by adenosine influences AHR, inflammatory cell influx, and airway remodeling. In aim 1 we will investigate the pro- inflammatory role of A3 receptors on mast cells in AHR, airway inflammation, and remodeling. In aim 2 we will investigate the capacity of agonist-induced activation of Gs-coupled adenosine receptors to limit AHR, airway inflammation, and remodeling. In aim 3 we will investigate constitutive activity of the A2B receptor. For all aims we will conduct in vitro experiments with human mast cells as well as mechanistic in vivo experiments using a series of models lacking adenosine receptors on mast cells. Completion of these aims will define pro- vs. anti-inflammatory signaling pathways on the mast cell, and identify the mechanisms by which adenosine- induced mast cell activation contributes to the cardinal features of asthma. PUBLIC HEALTH RELEVANCE. Asthma is a common chronic disease affecting approximately 10% of people in the United States. A better understanding of the inflammatory mediators involved in this disease, such as adenosine, will help identify new avenues of therapy, leading to better treatments for asthma
Project start date: 2002-12-01
Project end date: 2012-06-30
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Grants awarded to Stephen Lloyd Tilley
ADENOSINE RECEPTORS IN ALLERGIC INFLAMMATION
Stephen Lloyd Tilley
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5K08HL004280-05 from National Heart, Lung, And Blood Institute, IRG: ZHL1
Abstract: To successfully pursue biomedical research into the mechanisms of immune mediated lung injury, one must have a solid understanding of modern techniques in molecular biology, immunolgy, and pulmonary physiology. My training in pulmonary medicine has given me the opportunity to extensively study pulmonary physiology while diagnosing and treating patients with a variety of lung diseases. While I have begun to acquire some skills in molecular biology, an extended period of continued mentored training is critical for the development of the experience and skills necessary to pursue high quality research into lung disease. Dr. Koller s expertise in the generation and characterization of mouse models to study the pathogenesis of disease makes her laboratory an ideal environment for this mentored training. The overall objective of this project is to determine the contribution of adenosine to allergic airways disease by utilizing mouse models in which the expression of the A2b and A3 adenosine receptor genes have been inactivated using homologous recombination in embryonic stem cells. Specifically, we will define the expression of these receptors on immune cells and in murine lung, determine the receptor responsible for adenosine-induced mast cell degranulation, and determine the significance of activation of these receptors by adenosine to the overall pathogenesis of allergic airways disease. This project will provide extensive hands-on training in state of the art molecular biology, immunology, and murine pulmonary physiology in an environment that will promote the development of independent research skills necessary for a career investigating the mechanisms of pulmonary disease.
Keywords: hypersensitivity, inflammation, pathologic process, purinergic receptor, respiratory disorder, gene expression, mast cell, receptor expression, disease /disorder model, embryonic stem cell, laboratory mouse
Project start date: 2000-05-05
Project end date: 2005-04-30
5K08HL004280-05 (2004): $125982
5K08HL004280-04 (2003): $125982
5K08HL004280-02 (2001): $122154
1K08HL004280-01 (2000): $110160
Role Of Propionibacteria In Sarcoidosis
Stephen Lloyd Tilley
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5R21HL077581-02 from National Heart, Lung, And Blood Institute, IRG: ZHL1
Abstract: The objective of this proposal is to determine if Propionibacteria are etiologically important to the development of sarcoidosis in the United States (Aim 1), and to develop a mouse model of sarcoidosis using this bacterium (Aim 2). Several lines of evidence suggest that this anaerobic bacteria may be involved in the pathogenesis of disease. First Propionibacteria have been identified by culture and PCR in a high percentage of sarcoid lymph nodes from Japanese and European patients, and studies with in situ molecular probes detected Propionibacteria within sarcoid granulomas. Second, antibiotics effective against this organism showed remarkable efficacy in a small group of patients with cutaneous sarooidosis. Finally, animals exposed to this bacterium develop granulomatous inflammation similar to sarcoidosis. In Aim 1 we will look for evidence of Propionibacteria by performing quantitative PCR, with probes specific for this organism, on DNA from granulomas obtained by laser capture microscopy from parffin-embedded samples. We will also look for Propionibacteria by in situ hybridization. Finally, we will attempt to culture the organism from tissue obtained from patients undergoing diagnostic evaluation. Cultured bacteria will be characterized from patients with acute-resolving and chronic-persistent disease to see if the bacterial factors contribute to clinical phenotype. Cultured bacteria will also be used for inoculation in mice to produce an animal model. The initiation, maintenance, and resolution of granulomatous inflammation in mice of several different strains will be assessed following exposure to Propionibacteria. These experiments will allow us to look at host factors as determinants of clinical phenotype. Multiple organs will be evaluated including the lungs, lymph nodes, liver, spleen, heart, brain, and eyes, and skin to determine whether host factors or bacterial factors are important for specific organ involvement. Future directions will include a placebo-controlled trial of tetracyclines for sarcoidosis if Propionibacteria are identified.
Keywords: Propionibacterium, disease /disorder etiology, host neoplasm interaction, lymph node neoplasm, pathologic process, sarcoidosis, disease /disorder model, model design /development, transfection /expression vector, clinical research, human subject, in situ hybridization, laboratory mouse, laser capture microdissection, microorganism culture, polymerase chain reaction
Project start date: 2004-09-01
Project end date: 2006-06-30
5R21HL077581-02 (2005): $219000
1R21HL077581-01 (2004): $146000
Role Of Adenosine In Allergic Lung Disease
Stephen Lloyd Tilley
Medicineuniversity Of North Carolina Chapel Hill
Grant 2R01HL071802-05A1 from National Heart, Lung, And Blood Institute, IRG: LCMI
Abstract: Elevated adenosine levels in the lungs and exhaled breath of asthmatics, with further increases following antigen challenge, suggests that this ubiquitous mediator may contribute to the pathophysiology of asthma. Many of the effects of adenosine in the asthmatic lung are mast cell-dependent. Adenosine has both pro- and anti-inflammatory effects on mast cells, due to the expression of multiple adenosine receptors on the cell surface, each capable of activating very different intracellular signaling pathways. In this proposal we will test the hypothesis that both pro- and anti-inflammatory signals are transmitted to the mast cell by adenosine via the activation of distinct cell-surface receptors, and that engagement of these receptors by adenosine influences AHR, inflammatory cell influx, and airway remodeling. In aim 1 we will investigate the pro- inflammatory role of A3 receptors on mast cells in AHR, airway inflammation, and remodeling. In aim 2 we will investigate the capacity of agonist-induced activation of Gs-coupled adenosine receptors to limit AHR, airway inflammation, and remodeling. In aim 3 we will investigate constitutive activity of the A2B receptor. For all aims we will conduct in vitro experiments with human mast cells as well as mechanistic in vivo experiments using a series of models lacking adenosine receptors on mast cells. Completion of these aims will define pro- vs. anti-inflammatory signaling pathways on the mast cell, and identify the mechanisms by which adenosine- induced mast cell activation contributes to the cardinal features of asthma. PUBLIC HEALTH RELEVANCE. Asthma is a common chronic disease affecting approximately 10% of people in the United States. A better understanding of the inflammatory mediators involved in this disease, such as adenosine, will help identify new avenues of therapy, leading to better treatments for asthma
Project start date: 2002-12-01
Project end date: 2012-06-30
5R01HL071802-04 (2007): $311478
5R01HL071802-03 (2006): $320781
5R01HL071802-02 (2005): $328500
1R01HL071802-01A1 (2004): $182500
Stephen Lloyd Tilley
University Of North Carolina Chapel Hill
office Of Sponsored Research
chapel Hill, Nc 27599
Grant 1U19AI077437-018657 from National Institute Of Allergy And Infectious Diseases, IRG: ZAI1
Project start date: 2008-03-01
Project end date: 2013-02-28