Leukotriene E4 receptors in asthma pathogenesis

Lead Research Organisation: King's College London
Department Name: Medicine School Office

Abstract

The incidence of bronchial asthma has increased during last two decades in the UK, affecting more than 5 million people and putting the UK in the first place among European countries with the highest prevalence of the disease. In spite of great efforts we still do not fully understand the primary mechanisms leading to development of asthmatic symptoms. Cysteinyl leukotrienes (cysLTs) appear to be one of the major substances causing asthma and asthmatic lungs are very sensitive to the effects of these substances, especially to leukotriene E4 (LTE4). This disproportionate sensitivity to LTE4 is even more increased in a subpopulation of patients with aspirin-sensitive asthma. We propose to characterise new receptors for cysLTs and define the way that cysLTs exert their action on lung cells. Our research will allow an understanding of the role of the cysLTs in the mechanism of bronchial asthma and may lead to new treatments.

Technical Summary

Basic and clinical studies have indicated that cysteinyl leukotrienes (cysLTs) are one of the major proinflammatory mediators involved in the pathophysiology of bronchial asthma. Although leukotrienes have been known for more than 20 years we still do not fully understand their signalling and functions in pathological processes such as bronchial asthma.
Asthmatic airways respond with enhanced bronchoconstriction to inhaled cysLTs and especially to leukotriene E4 (LTE4) in comparison with normal subjects. This disproportionate hyperresponsiveness to LTE4 is even more increased in a subpopulation of asthmatics (about 10% of asthmatics), with aspirin-sensitive asthma. LTE4 has also been shown to be the most potent cysLT in eliciting influx of inflammatory cells such as eosinophils and basophils into bronchial mucosa of asthmatic subjects. None of these phenotypes can be explained based on current knowledge of cysLT signalling through their specific receptors. The existence of separate LTE4 receptors has been recently postulated. The P2Y12 has been suggested to be a novel LTE4 receptor in human cells and another, as yet unidentified LTE4 receptor has been described in CysLT1/CysLT2 double knock-out murine model. Our goal is to characterise the P2Y12 as the LTE4 specific receptor in human primary cells, define the human P2Y12 gene structure, including promoter and alternatively spliced transcript expression and mechanisms responsible for regulation of the receptor expression. To address whether P2Y12 may have a role in asthma pathogenesis, the receptor expression at the mRNA level as well responsiveness to LTE4 will be studied in cells derived from asthmatic patients in comparison to healthy control subjects. The role of currently used P2Y12 inhibitors as a potential, new anti-asthmatic drugs will be evaluated in in vitro models. Clinical data showing increased LTE4 responsiveness in asthmatic patients strongly suggest that an additional LTE4 receptor is expressed on airway smooth muscle cells, apart from P2Y12, with enhanced expression in asthmatics and with the highest expression present in bronchi of aspirin sensitive subjects. In order to identify this novel LTE4 receptor we will characterise the pattern of G-protein coupled receptor (GPCR) expression and LTE4-induced signalling in cells from asthmatics with different asthma phenotypes and healthy controls using real-time PCR array and calcium mobilization assay. Identified genes will be cloned, expressed heterologously and analysed for LTE4 responsiveness.
This project presents a unique opportunity to provide important insights on the role of LTE4 in asthma and the possibility of identifying new molecular targets for patient benefit.
 
Description Advances in mechanisms of allergy and clinical immunology in 2012
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in clinical reviews
 
Description IUPHAR review
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in systematic reviews
 
Description Asthma UK Travel Award
Amount £500 (GBP)
Organisation Asthma UK 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 05/2011 
End 09/2011
 
Description Asthma UK Travel Award
Amount £500 (GBP)
Organisation Asthma UK 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 08/2009 
End 09/2009
 
Description BRC Early Career Awards
Amount £4,400 (GBP)
Organisation King's College London 
Department NIHR Biomedical Research Centre
Sector Academic/University
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 11/2012 
End 03/2013
 
Description BSI travel award
Amount £500 (GBP)
Organisation British Society For Immunology 
Sector Learned Society
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 05/2013 
End 06/2013
 
Title Airway smooth muscle cell lines from healthy and asthmatic individuals 
Description Airway smooth muscle cell lines grown from endobronchial biopsies from healthy and asthmatic individuals. 
Type Of Material Cell line 
Year Produced 2011 
Provided To Others? Yes  
Impact Development of human primary cell model used for in vitro research on airway smooth muscle cells. This model is used for predicting airway responses in health and disease states. It may replace animal models for some experiments. 
 
Description Airway smooth muscle culture from bronchial biopsies 
Organisation King's College London
Department Department of Physiology
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Setting the method for in vitro culture of human airway smooth muscle from bronchial biopsies
Collaborator Contribution Selection of patients, performing bronchoscopies and collecting biopsies. Culture of human airway smooth muscle
Impact Setting up a unique model to study human airways. Research projects in progress.
Start Year 2009
 
Description Airway smooth muscle culture from bronchial biopsies 
Organisation King's College London
Department Department of Respiratory Medicine and Allergy
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Setting the method for in vitro culture of human airway smooth muscle from bronchial biopsies
Collaborator Contribution Selection of patients, performing bronchoscopies and collecting biopsies. Culture of human airway smooth muscle
Impact Setting up a unique model to study human airways. Research projects in progress.
Start Year 2009
 
Description Collaboration with Critical Care Medicine Department, NIH, Bethesda, USA 
Organisation National Institutes of Health Clinical Center
Department Critical Care Medicine Department (CCMD)
Country United States of America 
Sector Hospitals 
PI Contribution Project design, collaboration on experiments and data analysis. PMID: 21605126
Collaborator Contribution Research project on leukotriene signalling in human primary cells
Impact Publication: Chen LY, Eberlein M, Alsaaty S, Martinez-Anton A, Barb J, Munson PJ, Danner RL, Liu Y, Logun C, Shelhamer JH, Woszczek G.Cooperative and redundant signaling of leukotriene B4 and leukotriene D4 in human monocytes.Allergy. 2011 Oct;66(10):1304-11. PMID: 21605126
Start Year 2008
 
Description GPCR Imperial 
Organisation Imperial College London
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Collaboration on research on G-protein coupled receptors in immune responses. PMID: 22391114
Collaborator Contribution Collaboration on research on G-protein coupled receptors in immune responses. -Intelectual input -Access to facilities
Impact PMID: 22391114
Start Year 2010
 
Description GPCRs King's 
Organisation King's College London
Department Division of Asthma, Allergy and Lung Biology
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Collaboration on several projects studying role of G-protein coupled receptors in asthma and allergy. PMID: 23472176; PMID: 22391114
Collaborator Contribution Collaboration on several projects studying role of G-protein coupled receptors in asthma and allergy. -Intellectual input -Access to facilities -Training of staff
Impact PMID: 23472176; PMID: 22391114
Start Year 2010
 
Description Leukotriene receptors 
Organisation University College London
Department UCL Cancer Institute
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Research collaboration on the project concerning leukotriene receptors.
Collaborator Contribution Principal investigators of the project on comparative primate methylome analysis.
Impact Publication
Start Year 2012
 
Description NIHR BRC 
Organisation King's College London
Department NIHR Biomedical Research Centre
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Collaboration - research projects on asthma and allergy.
Collaborator Contribution Access to clinical and research facilities. Training.
Impact PMID: 23472176; PMID: 22391114
Start Year 2009