Dean Sheppard
University Of California San Francisco
Project start date: 1976-07-01
Project end date: 2017-06-30
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Dean Sheppard
In Vivo Function Of Pulmonary Integrins
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R37HL053949-13 from National Heart, Lung, And Blood Institute IRG: ZRG1
Abstract: This competitive renewal application is to continue on-going studies of the functions of the epithelial integrin, alphavbeta6. Based on analysis of beta6 knockout mice, alphavbeta6 plays a critical role in regulating pulmonary responses to injury through spatially and temporally regulated activation of the cytokine, transforming growth factor beta (TGFbeta). Alphavbeta6-mediated TGFbeta-activation is required for induction of pulmonary edema and pulmonary fibrosis in response to bleomycin, and homeostatically regulates macrophage protease expression, protecting the lungs from protease-mediated emphysema. Lung injury increases expression of alphavbeta6, but multiple lines of evidence suggest that under resting conditions alphavbeta6 is expressed but does not activate TGFbeta, and that alphavbeta6-mediated TGFbeta-activation is itself "activated" by extracellular signals. The studies proposed in this application will address the critical question of how this process is regulated. Preliminary evidence suggests that known downstream targets of integrin signaling, the focal adhesion kinase (FAK) and the small GTPases, Rac1 and RhoA activate this pathway in a model culture system. Furthermore, IL-1beta, a toll-like/IL-1 receptor (TIR) ligand, activates this process in polarized alveolar and airway epithelial cells. We will therefore determine the range of TIR ligands involved, the roles of known components of the TIR signaling pathway, and how this pathway connects to FAK, Rac1 and RhoA. Preliminary results suggest that at least one structural alteration in the beta6 subunit, that mimics a mutant known to "activate" integrins in leukocytes and platelets, also enhances alphavbeta6-mediated TGFbeta-activation. We will therefore examine a series of beta6 mutants targeting each of the steps associated with conformation-dependent activation of integrins on non-adherant cells to determine the extent to which activation of alphavbeta6-mediated TGFbeta-activation mimics this process of "inside-out" conformational change of other integrins. Finally, we will utilize lines of mice already available to us expressing null mutations of components of the TIR signaling pathway, and mice we will generate predicted to either inducibly activate this pathway in alveolar epithelial cells or inducibly express maximally "activated" integrin, to examine the relevance of our in vitro findings to each of the in vivo roles we have identified for alphavbeta6. The proposed studies should provide important general insights into the mechanisms underlying affinity and avidity modulation of integrins in adherant cells. They should also identify important targets that could be used to design treatments to intervene in acute lung injury, pulmonary fibrosis and emphysema.
Keywords: alveolar macrophage, cytokine, integrin, lung disorder, protein structure function, respiratory epithelium, cell adhesion, cell proliferation, conformation, cytokine receptor, enzyme induction /repression, focal adhesion kinase, gene mutation, guanosinetriphosphatase, interleukin 1, ligand, pulmonary fibrosis /granuloma, receptor binding, toll like receptor, transforming growth factor, laboratory mouse, laboratory rat
Project start date: 1995-04-01
Project end date: 2009-03-31
5R37HL053949-13 (2007): $359123
5R01HL053949-12 (2006): $369849
5R01HL053949-11 (2005): $378750
2R01HL053949-10 (2004): $378750
Integrin-Mediated Development Of The Thoracic Duct
Dean Sheppard, Professor
Medicineuniversity Of California San Francisco
3333 California St., Ste 315
san Francisco, Ca 941430962
Grant 5R01HL064353-09 from National Heart, Lung, And Blood Institute IRG: LBPA
Abstract: This competitive renewal application is to continue on-going studies of novel functions of the integrin, alpha9beta1. Based on analysis of alpha9 knockout mice, alpha9beta1 plays a critical role in development of the thoracic duct and other lymphatic vessels. One clue to the mechanism by which this integrin might contribute to lymphatic development comes from our identification of the lymphangiogenic growth factors, VEGFC and VEGFD as putative alpha9beta1 ligands. During the current funding period a unique mechanism by which alpha9beta1 enhances cell migration was also identified, and this effect was shown to be mediated by specific sequences in the alpha9 cytoplasmic domain. A single protein, the enzyme spermine/spermidine acetyltransferase, was found to bind to the alpha9 cytoplasmic domain and to specifically modulate alpha9-dependent enhancement of cell migration. This application proposes to evaluate each of these clues in more detail. The significance of binding of the extracellular domains of alpha9beta1 to VEGFC and D will be examined by assessing cell migration, proliferation, and early steps in integrin and growth factor receptor signaling in response to recombinant forms of each growth factor in mock- and alpha9-transfected cells. The importance of co-ligation of the canonical receptor for these growth factors, VEGFR3 will be assessed by performing all of these studies in the presence or absence of co-expression of VEGFR3. The alpha9 expressing cells critical for lymphatic development will be evaluated in mice homozygous for a conditional alpha9 null allele that will be used to inactivate this gene in specific cell types that might contribute to developing lymphatics. Stable cell lines co-expressing wild type and mutant forms of alpha9beta1 along with wild type or mutant forms of SSAT will be used to map the interaction sites in each protein and determine the importance of SSAT binding and enzymatic activity for alpha9beta1-mediated enhancement of cell migration. In vitro binding of recombinant versions of each protein, co-immunoprecipitation and double staining immunofluorescence will be used to determine whether this interaction is direct, occurs in living mammalian cells and results in co-localization to informative cellular compartments. Finally, the in vivo significance of alpha9-mediated migration and interactions with SSAT will be determined utilizing mice expressing knock-in mutations of the alpha9 cytoplasmic domain specifically designed to eliminate enhanced migration and/or SSAT binding. The proposed studies should provide important information about the role this widely expressed integrin plays in lymphatic development and cell migration and could ultimately lead to the design of novel interventions in diseases affected by each of these processes
Keywords: acyltransferase, cell migration, developmental immunology, integrin, lymphatic circulation, protein protein interaction, protein structure function cell proliferation, cytoplasmic receptor, hematopoietic stem cell, phenotype, protein binding, protein isoform, receptor expression, recombinant protein, vascular endothelial growth factor genetically modified animal, immunofluorescence technique, immunoprecipitation, laboratory mouse, transfection
Project start date: 2000-02-01
Project end date: 2009-11-30
5R01HL064353-09 (2008): $323212
5R01HL064353-08 (2007): $323212
5R01HL064353-07 (2006): $332865
5R01HL064353-06 (2005): $340875
2R01HL064353-05 (2004): $340875
MECHANISMS OF INITIATION AND PERSISTENCE OF ALLERGIC ASTHMA
Dean Sheppard, Director, Lung Biology Center
University Of California San Francisco, 3333 California St., Ste 315, San Francisco, Ca 94143-0962
Grant 5U19AI077439-03 from National Institute Of Allergy And Infectious Diseases
Abstract: This Asthma and Allergic Diseases Center Grant application is an effort to continue a long-standing fruitful collaboration among 4 UCSF faculty members with a long-standing interest in the mechanisms underlying allergic airway inflammation and asthma. The central goals of this application are to determine critical mechanisms underlying the initiation and persistence of allergic airway inflammation and airway hyperresponsiveness. The application includes two projects that will utilize murine models and a third that will examine the relevance of molecular targets and pathways identified in these models to asthma prevalence, severity and drug responsiveness in humans. The Center Principal Investigator and leader of Project 1, Dean Sheppard, has identified critical roles for activation of transforming growth factor ? (TGF??) by two different integrins (?v?6 and ?v?8) in airway hyperresponsiveness in a chronic model of allergic asthma and in modulation of cognate immune responses. This proposal will take advantage of a series of lines of genetically modified mice to directly examine the role of TGF?? in these effects, to determine the critical cells types responsible, and to identify the molecular mechanisms underlying these responses. Project 2 is based on recent data from Richard Locksley, the project leader, that chitin, a prominent structural component of fungi, parasites and crustaceans, activates macrophages and primes innate immune cells for initiation of type 2 immune responses. Work proposed in this project will utilize a series of novel reporter lines to examine the critical chitin-responsive cells and the mechanisms and relevance of this pathway for initiation of allergic inflammation. Project 3, co-led by Esteban Burchard and John Fahy will follow-up on preliminary observations about genetic associations between sequence variants in chitin-degrading enzymes and TGF?? and allergic sensitization and asthma to more deeply interrogate associations and gene-gene interactions for sequence variants in chitinases and multiple components of the TGF? activation and signaling pathways, including the integrin subunits examined in Project 1. This project will also evaluate the functional significance of associated genes using bronchoalveolar lavage samples and tissue from asthmatic patients and healthy control subjects. These projects will be supported by a Physiology and Tissue Analysis core that will provide extensive support for all 3 projects, and by a centralized Administrative core. Lay summary - This Center will evaluate the mechanisms underlying initiation and persistence of asthma. By identifying novel pathways and molecular targets and testing their relevance to asthma, asthma severity and drug response in humans, the work in the Center should provide clues for the development of new treatments for this common and often devastating disease. PROJECT 1 ?v Integrins in Cognate Immunity and Airway Hyperresponsiveness (SHEPPARD, D) PROJECT 1 Mice lacking the epithelial integrin, ?v?6, that we have shown activates latent TGF-?, are protected from the persistent airway hyperresponsiveness (AHR) that follows chronic allergen challenge. Surprisingly, this protection is not associated with any decrease in sub-epithelial airway fibrosis, a central TGF-? -dependent feature of this model. Mice with leukocyte specific knockout of the related integrin, ?v?8, which also activates TGF-? show evidence of enhanced adaptive immunity. In this proposal, we will determine whether these altered responses in ?v?6 subunit knockout mice are a direct consequence of loss of the ?v?6 integrin and/or of TGF-? activation from conducting airway epithelial cells using "rescue" mice expressing either the wild type integrin or active TGF-? in airway epithelial cells. We will evaluate the effects of ?v?6 antibodies and a TGF-? RII-lg chimera on these same endpoints to further confirm the importance of this pathway and evaluate the feasibility of targeting this pathway for therapeutic intervention. To determine the mechanisms by which loss of ?v?6 protects from induction of AHR, we will evaluate the relationship between airway responsiveness and expression of a small number of candidate genes identified as linked to this phenotype in preliminary experiments utilizing expression microarrays. We will also determine the cellular distribution of expression by immunostaining and/or in situ hybridization, and will evaluate functional significance using commercially available lines of mice expressing null mutations of specific candidates. Because the cytokine IL-13 is known to play a central role in induction of AHR in multiple models, and because two of the most promising candidates identified by microarrays, leukotrienes C4 synthase and interleukin-18 have been suggested to be upstream of IL-13 induction in the airways, we will also examine the cellular sources of IL-13 in chronically challenged wild type and ?v?6 knockout mice. Finally, we will determine how loss of leukocyte ?v?8 leads to enhancement of adaptive immunity and examine the relevance of this pathway to allergic airway inflammation and its consequences. Lay summary - This project will examine how a single growth factor, transforming growth factor ?, can either contribute to development of chronic asthma or inhibit allergic sensitization and its consequences, depending on where and how this growth factor is activated
Project start date: 2008-04-01
Project end date: 2013-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
PFA/PA: RFA-AI-07-002
5U19AI077439-03 (2010): $1555189
Sponsored Links Excellgen http://Excellgen.com
5U19AI077439-02 (2009): $1522770
3U19AI077439-02S1 (2009): $677883
ROLE OF INFLAMMATION OF INDUCED AIRWAY HYPERREACTIVITY
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01HL033259-08 from National Heart, Lung, And Blood Institute IRG: RAP
Abstract: This application is part of a long-term effort to determine the relationship(s) between airway inflammation and airway hyperresponsiveness, a central feature of asthma. The proposed experiments will focus on the role of tachykinins, a class of low molecular weight peptides that are present in the airways of several mammals, including humans. Tachykinins play an important role in several inflammatory processes and have recently been found to be required for the induction of airway hyperresponsiveness by toluene diisocyanate (TDI), a widely used chemical that is a well known cause of occupational asthma. TDI exposure also potentiates the effects of exogenously administered tachykinins, at least in part by inhibition of neutral endopeptidase, an enzyme that contributes to local tachykinin metabolism. In the proposed experiments, we will first determine whether stimuli known to induce release of tachykinins into the airways can themselves induce airway hyperresponsiveness in guinea pigs, and whether this effect requires simultaneous inhibition of neutral endopeptidase. In similar experiments we will evaluate the effects of exogenous administration of specific tachykinins. Next we will determine whether tachykinins contribute to the increase in airway responsiveness caused by two other inflammatory stimuli, ozone and inhaled antigen, by evaluating the effects of tachykinin antagonists, tachykinin depletion and neutral endopeptidase inhibition on these responses. We will also determine whether these stimuli, like TDI, themselves lead to inhibition of airway neutral endopeptidase activity. To determine the mechanisms by which tachykinins contribute to airway hyperresponsiveness we will evaluate the effects of tachykinins and of TDI in augmenting acetylcholine release from efferent nerves and in increasing airway epithelial permeability. We will also evaluate the role of platelet activating factor in these responses. In addition, we will evaluate the mechanisms by which TDI inhibits airway neutral endopeptidase activity, by evaluating the effects of TDI on purified neutral endopeptidase and on neutral endopeptidase activity in tracheal explants. In these experiments we will focus on possible roles of toxic oxygen metabolites and proteolytic enzymes in inactivating neutral endopeptidase. Finally, we will pursue any positive in vitro experiments with studies of neutral endopeptidase inactivation by in vivo exposure of guinea pigs to TDI. These experiments will help to clarify the role of an important class of inflammatory mediators (tachykinins) in the induction of airway hyperresponsiveness and will also further our understanding of the links between hyperresponsiveness and inflammation.
Keywords: asthma, inflammation, substance P, acetylcholine, antigen, environmental toxicology, lung, occupational disease, ozone, peroxide, platelet activating factor, pollution related respiratory disorder, protease inhibitor, proteinase, respiratory epithelium, respiratory hypersensitivity, respiratory pharmacology, stilbene 2, 4 diisocyanate, tachykinin, trachea, aerosol, guinea pig, tissue /cell culture
Project start date: 1985-09-20
Project end date: 1993-08-31
5R01HL033259-08 (1992): $165024
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5P50HL056385-059001 from National Heart, Lung, And Blood Institute
Abstract: The small animal core will provide a centralized facility for the measurement of pulmonary resistance and compliance in anesthetized and ventilated mice, and for the measurement of responsiveness to intravenous acetylcholine. The core will also assist in broncholeveolar lavage and in the inflation, perfusion, and embedding of mouse lungs. The core facility will be equipped with two complete plethysmographic systems, including plethysmographs, pressure transducers, amplifiers, pulmonary mechanics analyzers, and chart recorders, and will be staffed by a full time staff research associate under the direct supervision of the principal investigator. The core will be extensively utilized by 4 of the 6 projects in this proposal, and the existence of a core facility will insure both quality control and rapid dissemination of technical information among these 4 projects. By utilizing existing equipment, and a newly constructed, dedicated small animal laboratory, this centralized facility will obviate the needs to train additional personnel in each project lab, to duplicate expensive equipment, and to set aside space in each laboratory suitable for experiments with live animals.
Keywords: animal colony, biomedical facility, laboratory mouse, respiratory airway pressure, respiratory function, plethysmography
Sponsored Links Excellgen http://Excellgen.com
ROLE OF INFLAMMATION IN INDUCED AIRWAY HYPERREACTIVITY
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01HL033259-03 from National Heart, Lung, And Blood Institute IRG: RAP
Abstract: The proposed research is part of a long term effort to determine the mechanisms underlying the exaggerated bronchoconstrictor responsiveness that characterizes asthma. This project will combine methods of physiology, pharmacology and morphometry to study the mechanisms by which toluene diisocyanate (TDI), a widely used chemical that commonly causes asthma in exposed workers, induces both airway inflammation and airway hyperresponsiveness in guinea pigs. An advantage of this proposal is the long term collaborative interaction with a large group of scientists interested in the neurohumoral control of the lungs and airways. The general hypothesis we will test is that inflammation is a major mechanism of airway hyperresponsiveness. Thus, we will study the role of various aspects of the inflammatory response, including increased vascular permeability, chemical mediators of inflammation, and tissue infiltration with inflammatory cells, in the increase in airway responsiveness caused by exposure to TDI. First, we will use inhibitors of various steps in the metabolism of arachidonic acid to study the roles of prostaglandins, thromboxanes and leukotrienes in this response. By performing acetylcholine dose response curves and morphologic examination of sections of trachea and lung in treated and untreated animals exposed to air and to TDI, we will examine the effects of these inhibitors simultaneously on airway hyperresponsiveness and on morphologic evidence of airway injury and inflammation. Next, we will study the roles of polymophonuclear leukocytes (PMNs) in mediating the effects of TDI by performing similar studies in animals that have been depleted of PMNs or depleted and then repleted prior to exposure. We will perform similar studies in animals that have been depleted of eosinophils. In a parallel set of experiments we will study the role of inflammatory cells and mediators of inflammation in the increase in airway vascular permeability to Evans blue dye caused by exposure to TDI. Finally, we will study the effects of TDI on the in vitro behavior of airway smooth muscle both in the presence of and in the absence of PMNs eosinophils and airway epithelial cells. These studies should provide important insights into the mechanisms by which exposure to airborne chemicals can cause inflammation, edema, and hyperresponsiveness of the airways.
Keywords: BLOOD CELLS, LEUKOCYTES, NEUTROPHILS, CYANATES, ISOCYANATES, DISEASES, PATHOLOGIC PROCESSES, INFLAMMATION, FATTY ACIDS, EICOSANOIDS, ARACHIDONIC ACID, HYPERSENSITIVITY, RESPIRATORY HYPERSENSITIVITY, ASTHMA, PHENYLALKANES, TOLUENE, RESPIRATORY AND APPLIED PHYSIOLOGY STUDY SECTION, RESPIRATORY DISORDERS, ENVIRONMENTAL POLLUTANTS ASSOCIATED, ADRENAL CORTEX HORMONES, BENZOPYRROLE CARBOXYLIC ACIDS, INDOMETHACIN, BLOOD CELLS, LEUKOCYTES, EOSINOPHILS, CARDIOVASCULAR SYSTEM, ENDOTHELIUM PERMEABILITY, CHOLINE, ACETYLCHOLINE, FATTY ACIDS METABOLISM, EICOSANOIDS METABOLISM, FATTY ACIDS, EICOSANOIDS, LEUKOTRIENES, FATTY ACIDS, EICOSANOIDS, THROMBOXANES, HYPERSENSITIVITY, RESPIRATORY HYPERSENSITIVITY, MUSCLES, SMOOTH MUSCLE, NERVOUS SYSTEM, CRANIAL NERVES, VAGUS NERVE (10), OCCUPATIONAL HEALTH, OCCUPATIONAL DISEASES, RESPIRATORY DISORDERS, PULMONARY EDEMA, RESPIRATORY SYSTEM PHARMACOLOGY, RESPIRATORY SYSTEM, EPITHELIUM, RESPIRATORY SYSTEM, LUNG, RESPIRATORY SYSTEM, TRACHEA, TOXICOLOGY, ENVIRONMENTAL, DYES, ENDOCRINOLOGY, HORMONES INHIBITORS, FATTY ACIDS, EICOSANOIDS, PROSTAGLANDINS, ANTIPROSTAGLANDINAGENTS, MAMMALS, RODENTS, HYSTRICOMORPHA, GUINEA PIGS, NEUROSURGERY, DENERVATION (GENERAL), TISSUE (CELL) CULTURE
Project start date: 1985-09-20
Project end date: 1988-09-19
THE NHLBI-BAY AREA FUNCTIONAL GENOMICS CONSORTIUM
Dean Sheppard, Professor
Medicineuniversity Of California San Francisco
3333 California St., Ste 315
san Francisco, Ca 941430962
Grant 5U01HL066600-03 from National Heart, Lung, And Blood Institute IRG: ZHL1
Abstract: The NHLBI-Bay Area Functional Genomics Consortium will use gene-trap vectors to inactivate thousands of genes in mouse embryonic stem (ES) cells and make them freely available for the purpose of generating knockout mice. In preliminary studies, custom gene-trap vectors have been used to trap more than 500 mouse genes, some completely novel, and many corresponding to ESTs of unknown function. Approximately 150 of the "trapped" ES cell clones have been transmitted through the germline, and studies of the knockout mice have already led to the identification of completely novel genes that are important in cardiopulmonary development and disease. The Consortium involves several leading San Francisco Bay Area research institutions The J. David Gladstone Institutes, the University of California, San Francisco, and the University of California, Berkeley. The Consortium is organized into nine Components (1) Gene Trapping in Embryonic Stem Cells, (2) Computational Methods for Predicting Gene Function, (3) In Situ Hybridization, (4) Gene Expression Profiling and Analysis, (5) Mouse Resource for Pulmonary Disease, (6) Mouse Resource for Lipid Metabolism and Atherogenesis, (7) Mouse Resource for Cardiopulmonary Development, (8) Cardiopulmonary Genomics Education, and (9) Administration. The major objective of the Consortium (corresponding to Component 1) is to use custom gene-trap vectors to inactivate at least 2,500 genes per year in ES cells. Each "trapped" ES cell line will be posted on the Consortium´s website (genetrap.org) and will be distributed freely to the research community for the purpose of producing knockout mice. A second objective (corresponding to Components 2-4) is to assess which of the ES cell lines is likely to be valuable for understanding cardiopulmonary development and common cardiopulmonary diseases. To achieve this objective, the investigators will use computational approaches, expression profiling with DNA microarrays, and in situ hybridization studies. A third objective (corresponding to Components 5-7) is to select a few ES cell clones for the production of knockout mice, for the purpose of understanding genes involved in cardiopulmonary development and disease. The Consortium´s resources will be distributed freely to any interested investigator and should provide a catalyst for many different NHLBI-funded research programs
Keywords: embryonic stem cell, functional genomics, gene induction /repression, gene targeting blood lipoprotein metabolism, cardiopulmonary disease, cooperative study, developmental genetics, heart, in situ hybridization, lung, microarray technology, molecular biology information system, training biotechnology, clone cell, laboratory mouse, transgenic animal
Project start date: 2000-09-30
Project end date: 2004-07-31
5U01HL066600-03 (2002): $1626627
5U01HL066600-02 (2001): $1656822
1U01HL066600-01 (2000): $1690073
5U01HL066600-04 (2003): $1555968
IN VIVO FUNCTIONS OF PULMONARY INTEGRINS
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01HL053949-05 from National Heart, Lung, And Blood Institute IRG: LBPA
Abstract: The integrin alphavbeta6 is a receptor for the extracellular matrix proteins fibronectin and tenascin. This receptor is highly expressed in the epithelium of the respiratory tract and the skin during development, after injury or inflammation, and in malignant tumors, but is not expressed at either site in normal, healthy adults. In vitro, heterologous expression of alphavbeta6 in tumor cells increases cell adhesion to fibronectin and tenascin and also increases the capacity of these cells to proliferate. However, nothing is known about what role, if any, this receptor plays in vivo. To examine the in vivo function of alphavbeta6, we are producing mice that are deficient in receptor expression using standard homologous recombination in embryonic stem cells. In parallel, we are producing mice that overexpress either functional receptor, or a mutant version engineered to be incapable of binding ligand, under the control of highly active promoters that target expression to the airway epithelium (CC10), to alveolar type II cells and bronchiolar epithelium (SPC), or to basal keratinocytes (K14). Examination of the growth and development of mice deficient in beta6 protein should provide important clues to what role, if any, this receptor plays in epithelial differentiation and organogenesis. In addition, we propose to produce cutaneous wounds and respiratory epithelial injuries in these mice to determine the role of alphavbeta6 in epithelial repair in the lungs and skin. We also propose to examine the effects of overexpression of intact or mutant beta6, and of knockout of the beta6 gene, on the development of spontaneous and chemically-induced lung tumors. These studies should provide important information about the in vivo function of this integrin in lung (and cutaneous) development, in the repair of lung (and cutaneous) injury, and in the development, growth and/or progression of lung tumors.
Keywords: chemical carcinogenesis, embryogenesis, integrin, lung neoplasm, wound healing, cell differentiation, gene mutation, hyperplasia, keratinocyte, lung disorder, metastasis, respiratory epithelium, skin, skin disorder, laboratory mouse, transgenic animal
Project start date: 1995-04-01
Project end date: 2000-02-29
5R01HL053949-05 (1999): $280551
5R01HL053949-04 (1998): $271148
5R01HL053949-03 (1997): $261814
FUNCTION OF A NOVEL AIRWAY EPITHELIAL ADHESION RECEPTOR
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01HL047412-08 from National Heart, Lung, And Blood Institute IRG: LBPA
Abstract: The integrin alpha/v/beta6, a receptor for the extracellular matrix proteins fibronectin and tenascin, is highly expressed in both airway and alveolar epithelium during organogenesis and in response to injury or inflammation. Inactivation of the beta6 gene in mice leads to infiltration of the skin and airways with inflammatory cells, suggesting a role for this receptor in down-modulating epithelial inflammation. In vitro, alpha/v/beta6 augments the ability of epithelial cells to proliferate and modulates the expression of several epithelial-derived cytokines. The central goal of this proposal is to identify the signaling pathway or pathways by which alpha/v/beta6 induces these complex changes in cell behavior. This goal is approached through 4 specific aims 1) Native immunoprecipitation, western blotting, in vitro kinase assays, and in vitro binding assays with cell lysates from stable transfectants expressing intact or mutant alpha/v/beta6 will be used to determine the relationships between activation of known components of integrin-associated signaling complexes and subsequent alpha/v/beta6- induced changes in cell behavior. 2) Preliminary results suggest that alpha/v/beta6 inhibits activation of NF-kappaB, a family of transcription factors involved in regulation of cytokine gene expression. These results will be confirmed, and their relevance to transcription and cytokine gene expression will be assessed in cell lines stably transfected with intact or mutant beta6. 3) To identify the precise structural requirements for beta6-mediated cellular responses, proliferation, cytokine gene expression, and activation of specific signaling intermediates will be assessed in stable cell lines expressing a variety of mutant versions of beta6 containing deletions or point mutations within the C-terminal 11 amino acids. 4) Native immunoprecipitation, focal adhesion plaque isolation, affinity chromatography and the yeast two-hybrid system will be used to identify the proteins responsible for signaling specificity through alpha/v/beta6. By utilizing a variety of mutant versions of beta6 that do or do not support specific functions, it should be possible to rapidly assess the likelihood that each of the proteins thus identified is functionally significant. Together, these studies should provide new information about the mechanisms by which changes in the extracellular matrix can modulate the growth and differentiation of airway epithelial cells and modulate airway inflammation. Since epithelial remodeling and inflammation are important features of a number of diseases of the lungs and airways, this information could be important for understanding the pathophysiology of these diseases and for designing improvements in their treatment.
Keywords: cell growth regulation, integrin, protein structure /function, receptor binding, respiratory epithelium, cell adhesion, cell migration, cytokine, gene expression, inflammation, nuclear factor kappa beta, receptor expression, wound healing, affinity chromatography, immunoprecipitation, tissue /cell culture, transfection
Project start date: 1992-01-23
Project end date: 2000-05-31
5R01HL047412-08 (1999): $242987
Sponsored Links Excellgen http://Excellgen.com
5R01HL047412-07 (1998): $235327
5R01HL047412-06 (1997): $227186
MODULATION OF AIRWAY INFLAMMATION BY EPITHELIAL INTEGRINS
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5P50HL056385-050004 from National Heart, Lung, And Blood Institute
Abstract: The influx and activation of inflammatory cells in the airway wall is dependent, in large part, on the local release of inflammatory cytokines. Although much attention has focused on cytokines derived from leukocytes, it is now clear that airway epithelial cells, the most numerous Cell type in the airway wall, are themselves capable of synthesizing and secreting several cytokines that could profoundly influence airway inflammation. In non-epithelial cells, regulation of cytokine gene expression is critically dependent on signals provided from the extracellular matrix via the integrin family of transmembrane receptors. The existence of similar regulation of cytokine gene expression in airway epithelial cells could help to explain the coexistence of airway remodeling and persistent airway inflammation in asthma. We have recently generated lines of mice expressing a null mutation in the beta-6 integrin subunit, a subunit that is restricted in its expression to epithelia, especially in the lung and skin. Mice expressing this mutation develop and reproduce normally, but all have inflammatory cell infiltrates around hair follicles and scattered throughout the lungs and airways, consistent with a role for integrin- derived signals in regulation of inflammatory cell recruitment and/or activation in these sites. In the proposed studies, we will systematically examine the effects of various components of the normal extracellular matrix and of matrix proteins enriched in injured and inflamed airways, on the constitutive and stimulated synthesis and secretion of cytokines known to be synthesized by airway epithelial cells. Once we have determined the effects of various matrix proteins, we will examine the role of various integrins in these responses. For these experiments, we will use blocking anti-integrin antibodies, recombinant fragments of matrix proteins designed to be uniquely recognized by specific airway epithelial integrins, and murine airway epithelial cells derived from mice expressing mutations in integrin genes. Finally, to determine the significance of integrin and matrix-derived cytokine regulation in vivo, we will examine the influx of inflammatory cells, the production of selected cytokines, and the alterations in in vivo airway responsiveness produced when mice deficient in integrins or integrin ligands are challenged acutely or chronically by either immunologic or non-immunologic inflammatory stimuli. Through these studies, we hope to determine how alterations in airway epithelial integrins and/or their ligands could contribute to the chronic airway inflammation that characterizes persistent asthma.
Keywords: cytokine, gene expression, inflammation, integrin, respiratory epithelium, respiratory hypersensitivity, immunologic assay /test, laboratory mouse
Dean Sheppard, Director, Lung Biology Center
University Of California San Francisco, 3333 California St., Ste 315, San Francisco, Ca 94143-0962
Abstract: The administrative core will be responsible for all administrative activities of the Center and will insure that the Center operates according to its objectives and NIH guidelines. The core will be responsible for coordinating all fiscal mamagement and personnel management. The core will also coordinate and schedule regular monthly research conferences, Executive Committee meetings, visits by external advisors, meetings of internal advisors and an annual Center retreat. The core will also be responsible for the timely renewal and modification of animal research, bio-safety and radiation safety approvals, for insuring the timely training of all new and existing personnel in safe lab practices and for the coordination and preparation of the annual progress reports. The core will act as intermediary to all components of the program and to the funding agency to effectively administer and integrate all components of the program and to provide clear, effective and succinct communication. Lay summary - This core will provide the necessary administrative back-up to maintain effective communication among each of the projects and cores in this grant and between these projects and cores and groups of internal and external advisors and staff at the NIAID
Keywords: Allergic asthma; Animal Experimental Use; Animal Experimentation; Animal Research; Back; Budgets; Communication; Dorsum; Expenditure; Extrinsic asthma; Funding Agency; Funding Source; Grant; Guidelines; Human Resources; Manpower; Modification; NIAID; NIH; National Institute of Allergy and Infectious Disease; National Institutes of Health; National Institutes of Health (U.S.); Personnel Management; Preparation; Programs (PT); Programs [Publication Type]; Progress Reports; Radiation; Reports, Progress; Research; SCHED; Safety; Schedule; Training; United States National Institutes of Health; Visit; atopic asthma; conference; extrinsic allergic asthma; meetings; personnel; programs; ray (radiation); symposium
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5U19AI077439-03_9001 (2010): $196785
3U19AI077439-02S1_9001 (2009): $85850
5U19AI077439-02_9001 (2009): $192852
INTEGRIN-MEDIATED REGULATION OF AIRWAY SMOOTH MUSCLE
Dean Sheppard
Department/ Educational Institution Type:
Grant 1R01HL102292-01A1 from National Heart, Lung, And Blood Institute
Abstract: The a9¿1 integrin is highly expressed in airway smooth muscle. Mice we have generated lacking this integrin only in smooth muscle cells have marked in vivo airway hyperresponsiveness and lung slices from these mice have increased airway narrowing. We have previously shown that the a9 subunit cytoplasmic domain directly binds the enzyme spermine/spermidine acetyltransferase (SSAT), the rate limiting step in catabolism of higher order polyamines, and that this association and polyamine catabolism are important modulators of a9¿1 function. Pharmacologic stabilization of SSAT also augments a9¿1-dependent prevention of airway smooth muscle contraction. In the current application we will systematically evaluate the effects of a9¿1 on responses of airway smooth muscle to multiple contractile agonists and to isoproterenol-induced relaxation using 4 parallel experimental systems (in vivo AHR, tracheal ring contraction, airway narrowing in lung slices and shortening of airway smooth muscle cells). We will utilize a variety of mutant and chimeric constructs of a9 and SSAT, in vivo and in vitro studies with SSAT knockout mice, catalytically inactive SSAT mutants and genetic and pharmacologic inhibitors to thoroughly examine what contribution interaction of a9 with SSAT makes to this response. Force generation in smooth muscle depends on calcium-dependent actin-myosin cross-bridging and parallel actin polymerization, and we will systematically evaluate the effects of a9¿1 on each of these pathways. Because the two major cytosolic effects of higher order polyamines are prevention of potassium efflux through Kir channels and activation of the lipid kinase, PIP5K1?, we will pay special attention to the roles of Kir channels and PIP5K1? in this process. The proposed studies will test the overall hypothesis that ligated a9¿1 normally serves as a brake on airway narrowing by concentrating SSAT, catabolizing polyamines and thus inhibiting potassium efflux and/or PIP2 production, resulting in reduced calcium oscillations, decreased actin-myosin cross-bridging and/or impaired actin polymerization. Abnormalities in this pathway, either acquired or genetic, could contribute to diseases such as asthma that are characterized by enhanced airway narrowing. Project Exaggerated airway narrowing is a central feature of asthma. We have found that mice lacking a specific protein (an integrin) only in smooth muscle have exaggerated airway narrowing. In this proposal we will determine how this integrin normally prevents airway narrowing, a process that could be perturbed in diseases such as asthma
Keywords: 1, 4-Butanediamine; 1, 4-Butanediamine, N, N`-bis(3-aminopropyl)-; 1, 4-Butanediamine, N-(3-aminopropyl)-; 1, 4-Diaminobutane; 1D8 antigen; 4-(1-Hydroxy-2-((1-methylethyl)amino)ethyl)-1, 2-benzenediol; ing; Actins; Adenosine Triphosphatase, Myosin; Adhesion Plaques; Adhesions; Agonist; airway hyper-reactivity; Airway Hyper-responsiveness; airway hyperreactivity; airway hyperresponsiveness; airway smooth muscle; Antigenic Surface Determinant Protein OA3; Asthma; ATPase, Actin-Activated; Attention; Band 2 Protein; Binding; Binding (Molecular Function); biological signal transduction; Blood Coagulation Factor IV; Bronchial Asthma; Ca++ element; Calcium; Calcium Oscillations; Catabolism; CD47 Antigen; CD47 Antigen (Rh-Related Antigen, Integrin-Associated Signal Transducer); CD47 Glycoprotein; Cell Communication and Signaling; Cell Locomotion; Cell Migration; cell motility; Cell Movement; Cell Signaling; Cell-Matrix Adherens Junctions; Cellular Migration; Coagulation Factor IV; Complex; cSAT transferase; Cytoplasmic Domain; Cytoplasmic Tail; diamine acetyltransferase; Diamine N-Acetyltransferase; Disease; disease/disorder; Disorder; EC 2.3.1.57; EC 2.7; Enzymes; Extracellular Matrix, Integrins; Factor IV; Focal Adhesions; Focal Contacts; gene product; Generations; Genetic; heavy metal lead; heavy metal Pb; Human; Human, General; IAP-50 antigen; In Vitro; in vivo; inhibitor; inhibitor/antagonist; Integrin-Associated Protein; integrin-associated protein IAP, human; integrin-associated protein p50; Integrins; Intracellular Communication and Signaling; Isoprenaline; Isopropyl Noradrenaline; Isopropylarterenol; Isopropylnoradrenaline; Isopropylnorepinephrine; Isoproterenol; Isuprel; K element; Kinases; Knockout Mice; Lead; Leiomyocyte; Leukocyte Surface Antigen CD47; Link; Lipids; Lung; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; MER6; Mice; Mice, Knock-out; Mice, Knockout; Molecular Interaction; Motility; Motility, Cellular; Murine; Mus; Muscle Cell Contraction; Muscle Contraction; Muscle, Involuntary; Muscle, Smooth; Muscular Contraction; mutant; Myocytes, Smooth Muscle; Myosin Adenosinetriphosphatase; myosin ATP phosphohydrolase (actin translocating); Myosin ATPase; Myosins; Null Mouse; OA3; OA3 antigen; OVTL3 protein, human; pathway; Pathway interactions; Pb element; Phosphatidylinositol 4, 5-Biphosphate; Phosphatidylinositol 4, 5-Diphosphate; Phosphatidylinositol-4, 5-Bisphosphate; Phosphotransferases; PIP2; Play; Polyamine Catabolism; Polyamine Compound; Polyamines; polymerization; Potassium; prevent; preventing; Prevention; Process; Production; Proteins; PtdInsP2; PtIns 4, 5-P2; pulmonary; Putrescine; Putrescine Acetyltransferase; putrescine N-acetyltransferase; Regulation; Relaxation; respiratory smooth muscle; Respiratory System, Lung; response; Role; Sarcoplasmic Reticulum; SAT; screening; Screening procedure; screenings; Signal Transduction; Signal Transduction Systems; Signaling; Site; Slice; Smooth muscle (tissue); Smooth Muscle Cells; Smooth Muscle Myocytes; Smooth Muscle Tissue Cell; social role; Source; SPD/SPM acetyltransferase; Spermidine; Spermidine N1-Acetyltransferase; spermidine spermine acetyltransferase; spermidine-spermine N(1)-acetyltransferase; Spermidine/Spermine N1-Acetyltransferase; Spermine; SSAT; Surface Antigen Identified by Monoclonal Antibody 1D8; System; System, LOINC Axis 4; Talin; Testing; Tetramethylenediamine; thrombospondin-1 receptor CD47; Transphosphorylases; Two Hybrid; Waves, Calcium; Work; Yeast One Hybrid System; Yeast One/Two-Hybrid System; yeast two hybrid system; Yeasts
Relevance: Exaggerated airway narrowing is a central feature of asthma. We have found that mice lacking a specific protein (an integrin) only in smooth muscle have exaggerated airway narrowing. In this proposal we will determine how this integrin normally prevents airway narrowing, a process that could be perturbed in diseases such as asthma
Project start date: 2010-12-03
Project end date: 2014-11-30
Budget start date: 3-DEC-2010
Budget end date: 30-NOV-2011
PFA/PA: PA-10-067
1R01HL102292-01A1 (2011): $386250
Sponsored Links Excellgen http://Excellgen.com
Conference On Fibronectin, Integrins & Related Molecules
Dean Sheppard, Professor
Gordon Research Conferences West Kingston, Ri 02892
Grant 5R13CA097981-03 from National Cancer Institute IRG: ZCA1
Abstract: Research on cell interactions with extracellular matrices has exploded during the past decade. Fibronectin has been the prototype extracellular matrix molecule for much of this investigation and has been a major focus on this Gordon Conference since 1982. The discovery of integrins in the mid-1980s as receptors for fibronectin and other constituents of extracellular matrices expanded the scope of the conference. The emerging understanding over the last five years that plasma membrane and cytosolic molecular partners regulate integrin function and specificity has further expanded the scope of the conference. Together, fibronectin, integrins, and their binding partners control many basic biological processes including cell adhesion, cell shape, organization of the cytoskeleton and the extracellular matrix, cell motility, regulation of morphogenesis and tissue interactions, immune cell trafficking, regulation of cell activation state and response to growth factors, induction and resolution of inflammation, hemostasis, and wound repair. Important new insights have been made in recent years, including Determination of the first crystal structure of the extracellular domain of an integrin. This work provides a platform for detail molecular dissection of integrin function. Molecular mechanisms involved in regulation of the activation state of integrins, critical to their ability to bind ligand and transduce signals. The nature and regulation of signaling complexes assembled at adhesion sites, including cytoskeletal components, receptor and non-receptor tyrosine kinases, and other elements of signaling cascades. New families of ligands including growth factors, neuronal and immune adhesion molecules, and disintegrins, and new families of physically-associated molecules including growth factor and other receptors, tetraspanins, caveolin, and multiple cytosolic partners, that have broadened the context within which integrins function. A central role for integrin function and dysfunction in a variety of physiologic and pathologic states, which make these molecules focused therapeutic targets. The conference outlined in this application is directed at communicating these exciting new developments and stimulating discussion among participants from different disciplines. This cross-pollination is a most effective way of stimulating new waves of insight and information.
Keywords: fibronectin, integrin, meeting /conference /symposium, protein binding, protein protein interaction, protein structure function, receptor, travel
Project start date: 2003-01-17
Project end date: 2007-12-31
5R13CA097981-03 (2007): $10000
Dean Sheppard, Professor
Medicineuniversity Of California San Francisco
Grant 5R01AI024674-22 from National Institute Of Allergy And Infectious Diseases IRG: CMI
Abstract: Receptors for the Fc domain of IgG (FcgammaR) and for the third component of complement are critical for pathogen elimination in host defense against infection and in generation of an inflammatory response. In preliminary, we have found that specific functions of both FcgammaRs and the integrin complement receptor aMa2 are lost in dendritic cells and macrophages that lack glycan phosphatidylinositol (GPI) anchored proteins because of a targeted deletion of the enzyme, Pig-a, which is required for synthesis of the GPI anchor. GPI-anchored proteins are not uniformly distributed in the plasma membrane, but are found in lipid rafts, domains enriched in cholesterol and sphingolipids. In unactivated cells raft domains are small patches where, on the cytoplasmic face, some signaling molecules accumulate. Cell activation by a variety of means leads to lipid raft coalescence, which in turn may lead to activation of signaling cascades because of the increased concentration of signaling molecules in the enlarged raft domains. Ligated FcgammaRs and alphaMbeta2 can cluster together with aggregated lipid rafts, suggesting a potential role for GPI-anchored proteins in targeting these receptors to rafts. Raft association may be a critical step in transmembrane signaling by these receptors. The purpose of this application is to understand the molecular basis for the requirement for GPI anchored proteins in the function of FcgammaRs and aMa2, to determine the molecular mechanisms by which these receptors are targeted to lipid rafts, and to test the hypothesis that their targeting to the raft compartment allows communication of the ligated receptors with intracellular tyrosine kinase signaling cascades
Keywords: glycosylphosphatidylinositol, immunoglobulin G, immunoglobulin structure, integrin, leukocyte activation /transformation, protein protein interaction biological signal transduction, cell adhesion molecule, complement receptor, intracellular transport, leukocyte, macrophage, membrane activity, membrane structure, protein structure function, protein tyrosine kinase laboratory mouse, tissue /cell culture
Project start date: 1988-09-01
Project end date: 2010-02-28
5R01AI024674-19 (2006): $365005
ROLE OF A NEUTROPHIL INTEGRIN IN LUNG INFLAMMATION
Dean Sheppard, Professor
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01HL064353-04 from National Heart, Lung, And Blood Institute IRG: LBPA
Abstract: Neutrophils contribute to the tissue injury central to a number of common lung diseases, including acute lung injury, cystic fibrosis and chronic bronchitis. A key step in the development of neutrophil-mediated tissue injury is the recruitment of neutrophils to sites of extravascular injury. Although several of the critical receptors involved in the recruitment of neutrophils have been identified, considerable evidence suggests that unidentified receptors must participate in neutrophil recruitment, especially recruitment into the lung. Recently, we have identified the integrin alpha9beta1 on human neutrophils, and have found that this integrin together with its close structural relative, alpha4beta1, is critical for neutrophil migration across activated endothelial monolayers in vitro. The central issues addressed in this application are the mechanisms by which alpha9beta1 in the distinct steps of rolling, stable adhesion and endothelial transmigration, we will utilize blocking monoclonal antibodies and neutrophils derived from the bone marrow of alpha9 null chimeric mice. To determine the in vivo significance of alpha9beta1 on neutrophils we will examine neutrophil sequestration, extra-vascular emigration and neutrophil-mediated tissue injury in the lungs and peritoneal cavity in guinea pigs treated with alpha9beta1 blocking antibody and in chimeric mice with alpha9 null neutrophils. We will then utilize cell lines stably transfected with a variety of deletion and chimeric mutant versions of alpha9 to determine the role of specific sequences in the alpha9 and alpha4 cytoplasmic domains in adhesion, migration and endothelial transmigration. Finally we will utilize ch8imeric and mutant forms of alpha9 with defined functional properties to determine the roles of rapid spatial redistribution of integrins and cytoskeletal associations in integrin-mediated migration, both in leukocyte and non-leukocyte model systems. These studies should provide insight into key steps in neutrophil recruitment and could lead to the development of novel interventions for the treatment of diseases characterized by neutrophil-mediated tissue injury.
Keywords: inflammation, integrin, lung injury, neutrophil, cell adhesion, cell migration, vascular endothelium permeability, chimeric protein, laboratory mouse, tissue /cell culture, transfection, transgenic animal
Project start date: 2000-02-01
Project end date: 2003-11-30
5R01HL064353-04 (2003): $282740
5R01HL064353-03 (2002): $275925
5R01HL064353-02 (2001): $269310
1R01HL064353-01 (2000): $262886
AV INTEGRINA IN COGNATE IMMUNITY AND AIRWAY HYPERRESPONSIVENESS
Dean Sheppard, Director, Lung Biology Center
University Of California San Francisco, 3333 California St., Ste 315, San Francisco, Ca 94143-0962
Abstract: Mice lacking the epithelial integrin, avpe, that we have shown activates latent TGFp, are protected from the persistent airway hyperresponsiveness (AHR) that follows chronic allergen challenge. Surprisingly, this protection is not associated with any decrease in sub-epithelial airway fibrosis, a central TGFp-dependent feature of this model. Mice Iwith leukocyte specific knockout of the related integrin, avp8, which also activates TGFp show evidence of enhanced adaptive immunity. In this proposal, we will determine whether these altered responses in P6 subunit knockout mice are a direct consequence of loss of the avp6 integrin and/or of TGFp activation from conducting airway epithelial cells using "rescue" mice expressing either the wild type integrin or active TGFp in airway epithelial cells. We will evaluate the effects of avpe antibodies and a TGFpRII-lg chimera on these same endpoints to further confirm the importance of this pathway and evaluate the feasibility of targeting this pathway for therapeutic intervention. To determine the mechanisms by which loss of avp6 protects from induction of AHR, we will eavluate the relationship between airway responsiveness and expression of a small number of candidate genes identified as linked to this phenotype in preliminary experiments utilizing expression microarrays. We will also determine the cellular distribution of expression by immunostaining and/or in situ hybridization, and will evaluate functional significance using commercially available lines of mice expressing null mutations of specific candidates. Because the cytokine IL-13 is known to play a central role in induction of AHR in multiple models, and because two of the most promising candidates identified by microarrays, leuotriene C4 synthase and interleukin-18 have been suggested to be upstream of IL-13 induction in the airways, we willl also examine the cellular sources of IL- 13 in chronically challenged wild type and P6 knockout mice. Finally, we will determine how loss of leukocyte avp8 leads to enhancement of adaptive immunity and examine the relevance of this pathway to allergic airway inflammation and its consequences. Lay summary- This project will examine how a single growth factor, transforming growth factor p, can either contribute to development of chronic asthma or inhibit allergic sensitization and its consequences, depending on where and how this growth factor is activated
Keywords: Acute; Airway Hyper-responsiveness; Allergens; Allergic; Allergic asthma; Allergic inflammation; Aminoacetic Acid; Animal growth regulators, transforming growth factors; Antibodies; Antibodies, Blocking; Arg-Gly-Asp; Arginine; Arginine, L-Isomer; Arginine-Glycine-Aspartic Acid Cell Adhesion Domain; Aspartic Acid; Asthma; Basophilic Histiocyte; Basophils, Tissue; Binding; Binding (Molecular Function); Blocking Antibodies; Blood leukocyte; Body Tissues; Bronchial Asthma; Candidate Disease Gene; Candidate Gene; Cell Communication and Signaling; Cell Count; Cell Number; Cell Signaling; Chimera; Chimera organism; Chronic; Complex; Dendritic Cells; Dendritic cell activation; Development; Epithelial; Epithelial Cells; Exposure to; Extracellular Matrix, Integrins; Extrinsic asthma; Family; Fibrosis; GFAC; Gene Expression; Genes; Genetics, in situ Hybridization; Glycine; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; Human; Human, General; IFN-gamma-Inducing Factor; IGIF; IL-1 Gamma; IL-13; IL-18; IL-1g; IL13; IL18 Protein; IL1F4; INFLM; Immune Globulins; Immune response; Immunity; Immunoglobulins; Immunoglobulins / Antibodies; In Situ Hybridization; Inflammation; Integrins; Interferon-gamma-Inducing Factor; Interleukin 18 (Interferon-Gamma-Inducing Factor); Interleukin 18 Proprotein; Interleukin-1 Gamma; Interleukin-13; Interleukin-18; Interleukin-18 Precursor; Intracellular Communication and Signaling; Isoforms; Keratin; Knock-out; Knockout; Knockout Mice; L-Arginine; L-Aspartic Acid; LTC4 synthase; Lead; Leukocytes; Link; MGC12320; Maintenance; Maintenances; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow Mast Cell; Marrow leukocyte; Mediating; Mice; Mice, Knock-out; Mice, Knockout; Modeling; Molecular Interaction; Murine; Mus; Null Mouse; PAI-1; PAI1; PLANH1; Pathway interactions; Pb element; Peptides; Phenotype; Plasminogen Activator Inhibitor 1; Play; Population; Protein Isoforms; Proteins; RGD; RGD (peptide); RGD (sequence); RGD Cell Adhesion Domain; RGD Domain; RGD Motif; RGD Tripeptide Sequence; RGD tripeptide; Receptor Protein; Regulation; Reporter; Reticuloendothelial System, Leukocytes; Role; Serine or Cysteine Proteinase Inhibitor Clade E Member 1; Signal Transduction; Signal Transduction Systems; Signaling; Source; T memory cell; T-Cells; T-Lymphocyte; Testing; Tetracycline Antibiotic; Tetracyclines; Therapeutic Intervention; Thymus-Dependent Lymphocytes; Tissues; Transforming Growth Factors; Transgenic Organisms; Tumor Growth Factors; Type 1 Plasminogen Activator Inhibitor; Veiled Cells; White Blood Cells; White Cell; airway epithelium; airway epithelium infalmmation; airway hyper-reactivity; airway hyperreactivity; airway hyperresponsiveness; airway inflammation; airway remodeling; allergic airway disease; allergic airway epithelium inflammation; allergic airway inflammation; arginyl-glycyl-aspartic acid; atopic asthma; biological signal transduction; cytokine; experiment; experimental research; experimental study; extrinsic allergic asthma; gene product; heavy metal Pb; heavy metal lead; host response; immunoresponse; in situ Hybridization Staining Method; in vivo; intervention therapy; leukotriene A4-glutathione S-leukotrienyltransferase; leukotriene C4 synthetase; leukotriene-C4 synthase; mast cell; mastocyte; member; memory T lymphocyte; null mutation; pathway; receptor; research study; response; social role; thymus derived lymphocyte; transgenic; white blood cell; white blood corpuscle
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5U19AI077439-03_0001 (2010): $347988
3U19AI077439-02S1_0001 (2009): $151862
5U19AI077439-02_0001 (2009): $341137
Sponsored Links Excellgen http://Excellgen.com