Glucocorticoid regulation of interleukin 8 expression in chronic obstructive pulmonary disease

Lead Research Organisation: University of Manchester
Department Name: Medical and Human Sciences

Abstract

Chronic obstructive pulmonary disease (COPD) is an inflammatory lung condition and a major cause of deaths worldwide. Since it is an inflammatory disease antiinflamatory medication such as steroids should be effective. However steroids usually fail to improve the COPD disease course significantly. This clinically observed resistance to steroid treatment is likely to be important for understanding the basis of the disease itself, and also for the clinical management of patients.
We are, therefore, currently investigating steroid action in COPD. We are doing this by comparing inflammatory lung cells between COPD patients and healthy volunteers. Interleukin 8 is a critical signal for recruitment of inflammatory cells to the lung, and is expressed at high level in COPD. Interleukin 8 is typically inhibited by glucocorticoids; but this effect is lost in COPD, contributing to their lack of beneficial effect. We will discover why interleukin 8, in COPD, acquires resistance to glucocorticoid inhibition. This will indicate new approaches to COPD treatment.

Technical Summary

Background: Chronic obstructive pulmonary disease (COPD) is a major cause of premature death, and morbidity throughout the world. The disease is generally resistant to glucocorticoids (GC). Alveolar macrophages appear to have a central role, attracting neutrophils and lymphocytes. COPD encompasses many different phenotypes. A high sputum eosinophil count is associated with better glucocorticoid response therefore defining a particular disease phenotype. Interleukin 8 production by airway macrophages is constitutively high, and resistant to inhibition by GC, whereas peripheral blood derived macrophages respond to both stimulation with lipopolysaccharide (LPS), and inhibition with GC.

Aim: To define the basis for the constitutively high expression of IL-8.

Design: I will recruit patients with COPD, either with high sputum eosinophil counts or low eosinophil counts. Airway macrophages will be harvested at bronchoscopy, with airway mucosal biopsies.

Objective 1: I will define ex-vivo cytokine responses to stimulation with LPS, and inhibition with GC, p38 kinase or both. A limited immunohistochemical study will then be done using previously optimised antibodies to cells.

Objective 2: Using celltracker software and confocal microscopy I will measure expression, and activation of the glucocorticoid receptor (GR). I will also track p38 and NFkB. I will determine protein expression in conditions of LPS activation, p38 inhibition, GC treatment and inhibition with both p38 and GC.

Objective 3: Using a new form of chromatin immunoprecipitation. I will examine the IL8 gene architecture, and transcription factor binding. I will compare lung with peripheral blood derived cells, and will use the TNF gene, which is repressed by GC under the same conditions, as a further control.

Scientific and Medical Opportunities: I will define why IL8 is not repressed by GC therefore suggesting new therapeutic approaches. I will also be able to use p38 inhibitors, which are in early clinical trials, to investigate the role of this kinase in modulating GC action.

Publications

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Blaikley JF (2011) Management of peripheral lymph node tuberculosis in routine practice: an unselected 10-year cohort. in The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease

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Carter SJ (2016) A matter of time: study of circadian clocks and their role in inflammation. in Journal of leukocyte biology

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Gibbs JE (2012) The nuclear receptor REV-ERBa mediates circadian regulation of innate immunity through selective regulation of inflammatory cytokines. in Proceedings of the National Academy of Sciences of the United States of America

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Trebble P (2010) Familial glucocorticoid resistance caused by a novel frameshift glucocorticoid receptor mutation. in The Journal of clinical endocrinology and metabolism

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Trump RP (2013) Optimized chemical probes for REV-ERBa. in Journal of medicinal chemistry

 
Description Biomedical research centre microarray award
Amount £8,500 (GBP)
Organisation National Institute for Health Research 
Department NIHR Manchester Musculoskeletal Biomedical Research Unit
Sector Public
Country United Kingdom
Start  
 
Description Clinical lecturer starter grant
Amount £28,269 (GBP)
Organisation Academy of Medical Sciences (AMS) 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2012 
End 06/2013
 
Description Early career grant
Amount £9,600 (GBP)
Organisation Society for Endocrinology 
Sector Learned Society
Country United Kingdom
Start 02/2013 
End 02/2018
 
Description Eureopean Respiratory Society COPD travel award
Amount £800 (GBP)
Organisation European Respiratory Society (ERS) 
Sector Charity/Non Profit
Country European Union (EU)
Start  
 
Description MRC clinician scientist award
Amount £1,007,966 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 05/2014 
End 05/2018
 
Description PUMP priming award for new technology
Amount £9,500 (GBP)
Organisation National Institute for Health Research 
Sector Public
Country United Kingdom
Start  
 
Description clinician scientist
Amount £1,102,534 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 09/2014 
End 08/2019
 
Title THP-1 knockdown cells in which a clock protein has been knocked down 
Description THP-1 cell line in which the clock protein REVERBalpha was knocked down using shRNA. 
Type Of Material Cell line 
Provided To Others? No  
Impact Allowed us to confirm that IL-6 is regulated by REVERBalpha and this was subsequently published in a PNAS paper. 
 
Description Industry sponsorship 
Organisation GlaxoSmithKline (GSK)
Country Global 
Sector Private 
PI Contribution We provided preliminary data showing that a clock protein, REVERBalpha, regulated the immune system. We were able to investigate the accuracy of the ligands targetting REVERB alpha in primary cells suggesting their translational importance.
Collaborator Contribution GSK developed novel ligands and synthesised them, allowing us to use them in our investigations.
Impact The ligands have enabled us to gain two NIHR awards. The access to the ligands also was a key part of my clinician scientist application. Two papers: 1) Optimised chemical probes for REVERBalpha J Med Chem 2) The nuclear receptor REV-ERBa mediates circadian regulation of innate immunity through selective regulation of inflammatory cytokines. PNAS
Start Year 2008
 
Description Lung transplant lectureship 
Organisation Manchester University NHS Foundation Trust
Country United Kingdom 
Sector Public 
PI Contribution From my clinical research fellowship I was able to successfully apply for a clinical lectureship gaining data for my intermediate fellowship application
Collaborator Contribution My partners allowed me to gain samples whilst in the department.
Impact Three sets of peer reviewed funding came from my time in the department (Clinical lectureship grant, SFE early career grant and a MRC clinician scientist award)
Start Year 2011
 
Description British thoracic society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Paper Presentation
Geographic Reach National
Primary Audience Health professionals
Results and Impact I presented my data concerning the function of reverb alpha in macrophages whilst being awarded the BTS young investigator of the year.

Rev-erba, a novel anti-inflammatory target, modifies the circadian oscillation of pulmonary inflammation
J Blaikley, L Green, J Gibbs, S Farrow, A Loudon, D Singh, D Ray


This raised my profile within the respiratory community allowing me to set up national and international collaborations to support my clinician scientist application
Year(s) Of Engagement Activity 2010
URL http://thorax.bmj.com/content/65/Suppl_4/A1.1.abstract
 
Description European respiratory society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Poster Presentation
Geographic Reach International
Primary Audience Health professionals
Results and Impact European respiratory society

Reverba is a novel regulator of COPD macrophage inflammation and glucocorticoid resistance J. Blaikley, J. Gibbs, A. Loudon, S. Farrow, D. Singh, D. Ray

Interest was generated in clocks and this lead to future collaborations that otherwise would not have been possible
Year(s) Of Engagement Activity 2011
URL https://www.ersnetsecure.org/public/prg_congres.detail?ww_i_session=17878&ww_c_mode=TEXTE-SPECIAL
 
Description Toronto University 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The talk sparked interest in biological timing mechanisms

Collaboration with Richard Horner
Year(s) Of Engagement Activity 2014