Variants of mast cell tryptase as markers and mediators of inflammation and tissue remodelling in asthma

Lead Research Organisation: University of Southampton
Department Name: Inflammation Infection and Repair

Abstract

Asthma and other allergic diseases represent a major health problem in this country. These conditions are associated with the release into the lungs of substantial quantities of an enzyme called tryptase. This has become a new target for drug development.
There are multiple forms of tryptase, prominent among which are the alpha and beta forms. We propose to investigate the degree to which asthmatic patients may exhibit genetic differences in the tryptases they possess. The relatively little studied alpha tryptase will be purified and its properties compared with those of beta tryptase.
The pattern of release of tryptases from cells will be studied, and the extent to which they are released into blood or fluids lining the lung examined. We shall carefully investigate the ability of the different tryptases to alter the function of cells to promote inflammation and structural alterations in the lung. These studies should advance understanding of fundamental processes of allergic disease and on the new approaches to treatment directed against tryptases.

Technical Summary

Tryptase, the major product of human mast cells, is emerging as a key mediator of inflammation and tissue remodelling in asthma and other allergic conditions. This tetrameric serine protease is often considered as a single enzyme, but multiple sequences have been derived, and a family of related proteases mapped to chromosome 16p13.3. We shall investigate the degree of genetic variation at this site in families with bronchial asthma, and examine the potential for linkage with features of disease. Alpha tryptase will be purified from a recombinant expression system and enzymatic properties investigated with synthetic and natural substrates. Comparison will be made with the more extensively studied beta tryptases, and the degree to which heterotypic tetramers may form will be examined. The cellular expression and release will be examined of alpha and beta of tryptase in vitro, and in vivo in body fluids from patients with allergic conditions. The ability of alpha tryptase to stimulate altered gene expression in epithelial cells and airway smooth muscle cells will be investigated using microarray procedures, and compared with the patterns with beta tryptase. The release of selected cytokines, matrix components or mediators, and the rate of cell proliferation in response to alpha and beta tryptases will be investigated, exploring potential proteolytic and non-proteolytic mechanisms. The abilities of alpha tryptase to provoke pro-inflammatory changes following transfer to a guinea pig model will be assessed and compared with those of beta tryptase. This integrative biology approach should yield crucial information on the mediator actions of human tryptases and their potential as targets for therapeutic intervention.

Publications

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Abdelmotelb AM (2014) Alpha-tryptase gene variation is associated with levels of circulating IgE and lung function in asthma. in Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

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Bergqvist A (2015) Alveolar T-helper type-2 immunity in atopic asthma is associated with poor clinical control. in Clinical science (London, England : 1979)

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Bradding P (2006) The role of the mast cell in the pathophysiology of asthma. in The Journal of allergy and clinical immunology

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Buckley M (2008) Identification of mast cells and mast cell subpopulations. in Methods in molecular medicine

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De Nadaï P (2006) CCR5 usage by CCL5 induces a selective leukocyte recruitment in human skin xenografts in vivo. in The Journal of investigative dermatology

 
Description Anaphylaxis provoked by anaesthetics
Geographic Reach Multiple continents/international 
Policy Influence Type Membership of a guidance committee
Impact The guidelines will encourage the routine collection of blood samples for the measurement of mast cell tryptase.
 
Description Risk factors for anaphylaxis
Geographic Reach Multiple continents/international 
Policy Influence Type Membership of a guidance committee
Impact Increases awareness of anaphylaxis as a clinically important condition
 
Description Asthma UK Project Grant
Amount £148,000 (GBP)
Organisation Asthma UK 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2006 
End 11/2009
 
Description Educational grant
Amount £50,000 (GBP)
Organisation Novartis 
Sector Private
Country Global
Start 10/2012 
End 09/2015
 
Description European Union Cooperation in Science and Technology COST scheme
Amount € 10,000 (EUR)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 05/2012 
End 09/2012
 
Description FSA, Studentship
Amount £60,000 (GBP)
Organisation Food Standards Agency (FSA) 
Sector Public
Country United Kingdom
Start 11/2007 
End 11/2010
 
Description Flexibility and Sustainability Funding (FSF)
Amount £86,259 (GBP)
Organisation National Institute for Health Research 
Department Comprehensive Clinical Research Network (Coordinating Centre) – NIHR
Sector Public
Country United Kingdom
Start 10/2011 
End 09/2013
 
Description Graduate studentship
Amount £70,000 (GBP)
Organisation Bayero University Kano 
Sector Academic/University
Country Nigeria
Start 02/2015 
End 01/2019
 
Description Graduate studentship
Amount £70,000 (GBP)
Organisation Iraqi Government 
Sector Public
Country Iraq
Start 10/2014 
End 09/2018
 
Description Higher Education Innovation Funding
Amount £15,000 (GBP)
Organisation Higher Education Funding Council for England 
Sector Public
Country United Kingdom
Start 04/2014 
End 06/2014
 
Description Impact Acceleration Account
Amount £18,000 (GBP)
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 11/2013 
End 03/2014
 
Description Institute for Life Sciences/Faculty Studentship
Amount £60,000 (GBP)
Organisation University of Southampton 
Sector Academic/University
Country United Kingdom
Start 10/2014 
End 09/2017
 
Description Lullaby Trust Project grant
Amount £10,000 (GBP)
Organisation The Lullaby Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2015 
End 08/2015
 
Description NERC Environment and Human Health Programme NE/E009417/1 Plant virus infection as a determinant of pollen allergenicity
Amount £60,678 (GBP)
Funding ID NE/E009417/1 
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 07/2008 
End 06/2010
 
Description NERC Innovation Grant
Amount £81,000 (GBP)
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 05/2009 
End 12/2010
 
Description National Research Priority Funding
Amount $930,000 (USD)
Organisation Qatar Foundation 
Department Qatar National Research Fund
Sector Charity/Non Profit
Country Qatar
Start 12/2013 
End 09/2017
 
Description New and Emerging Applications of Technology Programme NEAT H039 Development of discriminating tests for allergic reactions to medicines and anaesthetics, based on the assay of unique products of mast cell and basophil activation
Amount £244,408 (GBP)
Funding ID H039 
Organisation National Institute for Health Research 
Sector Public
Country United Kingdom
Start 11/2006 
End 07/2010
 
Description Part funded studentship
Amount £21,000 (GBP)
Organisation University of Southampton 
Sector Academic/University
Country United Kingdom
Start 10/2012 
End 09/2015
 
Description Petroleum Technology Development Fund Scholarship
Amount £75,000 (GBP)
Organisation PTDF Petroleum Technology Development Fund Nigeria 
Sector Public
Country Nigeria
Start 01/2020 
End 12/2024
 
Description Research grant
Amount £8,937 (GBP)
Organisation Asthma Allergy & Inflammation Research 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2016 
End 09/2018
 
Description Santander Internationalisation Fund
Amount £1,000 (GBP)
Organisation Santander Universities 
Sector Private
Country United Kingdom
Start 04/2012 
End 06/2012
 
Description Scholarship
Amount £10,000 (GBP)
Organisation Government of Egypt 
Sector Public
Country Egypt
Start 04/2016 
End 10/2016
 
Description Wessex Medical Trust, States of Jersey Research Grant
Amount £45,000 (GBP)
Organisation States of Jersey 
Sector Public
Country Jersey
Start 05/2008 
End 09/2010
 
Title Assay for mast cell chymase 
Description Diagnostic assay 
Type Of Material Technology assay or reagent 
Year Produced 2010 
Provided To Others? Yes  
Impact Improved diagnosis 
 
Title Improved assay for mast cell tryptases 
Description New monoclonal antibodies have been prepared and incorporated into new prototype assays for tryptase 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Production of new monoclonal antibodies against tryptases has led to development of better assays for these proteases. 
 
Description Ahmed Abdelmotelb 
Organisation Tanta University
Department Faculty of Medicine
Country Egypt 
Sector Academic/University 
PI Contribution Provided training for PhD student over four years, and subsequently supervised as a research fellow
Collaborator Contribution Provided PhD student for 4 years
Impact Successful completion of PhD course Publications pending
Start Year 2006
 
Description Changzhou University, China 
Organisation Changzhou University
Country China 
Sector Academic/University 
PI Contribution Presentations at Changzhou University, arrangements for student supervision, collaborative research, joint grant applications.
Collaborator Contribution Travel funding to China, collaborative research, joint grant applications.
Impact One research paper published, and another in advanced preparation. A research grant awarded in China for the joint work, and another application will shortly be submitted.
Start Year 2015
 
Description Chaza Mecheri 
Organisation University of Paris XII: Paris-Val-de-Marne
Country France 
Sector Academic/University 
PI Contribution training in cell culture, studies with mast cell proteases
Collaborator Contribution training of student in research practice
Impact publication pending
Start Year 2012
 
Description Helmut Haas 
Organisation Borstel Research Centre
Country Germany 
Sector Private 
PI Contribution Graduate student from our lab conducted studies in the partner laboratories using reagents prepared by our group
Collaborator Contribution Provided training in new molecular biology procedures
Impact Studies conducted included in a PhD thesis Publication pending
Start Year 2009
 
Description Helmut Haas 
Organisation Borstel Research Centre
Country Germany 
Sector Private 
PI Contribution Graduate student from our lab conducted studies in the partner laboratories using reagents prepared by our group
Collaborator Contribution Provided training in new molecular biology procedures
Impact Studies conducted included in a PhD thesis Publication pending
Start Year 2009
 
Description Hirohisa Saito 
Organisation National Research Institute for Child Health and Development
Country Japan 
Sector Academic/University 
PI Contribution Supervised a PhD student for a period of one year supported by his institution in Japan
Collaborator Contribution Graduate student worked in our laboratory for one year
Impact Study presented at national meeting Publication pending
Start Year 2011
 
Description Luis Escribano 
Organisation Spanish Network Centre for Mastocytosis
Country Spain 
Sector Charity/Non Profit 
PI Contribution Development of improved methods for diagnosis and clinical modelling of mastocytosis.
Collaborator Contribution Provision of samples from well characterised cases of mastocytosis.
Impact publications
Start Year 2007
 
Description Mogib Khedr 
Organisation Suez Canal University
Department Faculty of Pharmacy
Country Egypt 
Sector Academic/University 
PI Contribution Provided training for a PhD student over 4 years
Collaborator Contribution Provided a PhD student for 4 years
Impact Successful completion of PhD course Several presentations at national and international meetings. One paper published and further publications pending
Start Year 2008
 
Description Novel inhibitors of tryptase as anti-allergic agents 
Organisation Centre for Ecology & Hydrology (CEH)
Country United Kingdom 
Sector Public 
PI Contribution Access to data and intellectual input into collaborator's research.
Collaborator Contribution Provision of novel reagents
Impact Grant funding from NERC
Start Year 2009
 
Description Plant virus-induced alterations in the allergenicity of pollen 
Organisation Centre for Ecology & Hydrology (CEH)
Country United Kingdom 
Sector Public 
PI Contribution Access to data and intellectual input into collaborator's research.
Collaborator Contribution Provision of research materials, and useful discussions
Impact Grant funding from two separate NERC-led initiatives. Research paper published
Start Year 2008
 
Description Sayed Goda 
Organisation Anti-Doping Laboratory Qatar
Country Qatar 
Sector Public 
PI Contribution Supervision of research fellow and three research assistants. Set up a split site graduate student programme.
Collaborator Contribution Provision of staff to work in our laboratory Provision of materials from their institution
Impact Grant award. Presentations at national and international meetings
Start Year 2013
 
Description Uli Blank 
Organisation Paris Diderot University
Country France 
Sector Academic/University 
PI Contribution training in techniques related to calcium flux and provision of laboratory facilities
Collaborator Contribution training in techniques of kidney cell culture
Impact New information on fundamental processes in kidney disease
Start Year 2012
 
Description Support to mastocytosis patients' support group - York 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Participants in your research and patient groups
Results and Impact As a researcher in the mast cell field I was approached to give a talk at a meeting which led to the foundation of a support group for mastocytosis suffers in the UK (first national meeting was in York). There has been frequent contact since then and I have advised subsequently on aspects of the condition, on research in the field, etc.

The group is growing in size and in influence, and is now exploring setting up a research charity.
Year(s) Of Engagement Activity 2007