MICA: Evaluation of anti diabetic drugs in the treatment of respiratory disease

Lead Research Organisation: St George's University of London
Department Name: Basic Medical Sciences

Abstract

The airways of the lung are lined with a thin layer of fluid (airway surface liquid, ASL) which is important for lung defence against infection. To optimise its function, the volume and composition of ASL are tightly regulated by the surface cells of the airway (epithelium). We have shown that the concentration of glucose (sugar) in ASL is normally much lower than that of blood. Our evidence indicates that this serves as a protective mechanism as glucose is a growth substrate for many organisms including infectious microbes. However, glucose concentration in ASL increases when the airways are inflamed, when blood glucose concentration is raised (hyperglycaemia, associated with diabetes or severe illness) and, more potently, when inflammation and hyperglycaemia are both present. This makes the airway more susceptible to infection particularly with pathogens such as methicillin resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa.

We have developed a cell model of human airway epithelium to understand how glucose concentration in ASL is regulated. We have shown that glucose applied to the basolateral (blood) side can get across the epithelium into the ASL. Normally the movement of glucose between the epithelial cells is restricted. However, when the epithelium becomes inflamed, it becomes leakier allowing glucose to pass across it more easily. If this is coupled with increased glucose concentration on the basolateral side (as in diabetes), even more glucose gets across. In this situation, we showed that the growth of bacteria in ASL was increased. In an exciting new development, we prevented the growth of bacteria by treating the epithelium with metformin (a drug already in clinical use). Metformin predominantly acted to prevent glucose getting into ASL and supporting bacterial growth. This could potentially provide a new therapeutic route for treatment of lung disease particularly in the light of increased resistance of bacteria to antibiotic therapy.

It is difficult to investigate how inflammation and diabetes increase glucose in ASL in the lungs of human subjects. Therefore, we will use our cell model and animal models of human lung disease to gain an understanding of how this occurs. We will investigate whether metformin can reduce glucose concentrations in ASL and suppress respiratory infection. We will also investigate whether other drugs that are predicted to reduce glucose movement into ASL will have a similar effect. In addition, we will test whether new drugs that are being developed to reduce blood sugar levels in diabetes have an additive beneficial effect. This project will increase our understanding of how glucose is increased in ASL in lung disease and when blood sugar levels are raised and how these two events promote infection. It will also tell us whether metformin or other drugs that reduce glucose movement across the epithelium or reduce blood glucose could be used to treat patients with these conditions. This could have important economic and social impact particularly in wealthy countries where the ageing population is expanding, chronic illness is more prevalent and the incidence of diabetes is increasing.

Technical Summary

The respiratory tract is lined with a thin layer of fluid (airway surface liquid, ASL), which is important for lung defence against infection. Glucose concentrations in ASL are normally 12x lower than that of blood. However, they are raised in people during airway inflammation, hyperglycaemia and, more potently, when both are present, making the airway more susceptible to infection from pathogens such as methicillin-resistant S.aureus and P.aeruginosa.

We have shown in airway epithelial cells in vitro that, under normal conditions, glucose diffuses from blood into ASL but accumulation is limited by the permeability of the epithelium, blood glucose concentration, glucose uptake into the epithelial cell via GLUT transporters and its metabolism. Inflammation increased transepithelial permeability to glucose and elevated glucose in ASL. In addition, increasing basolateral glucose concentration increased glucose in ASL and promoted the growth of S.aureus on the apical surface of lung epithelial cells. In vitro, pre-treatment of airway epithelia with metformin (a drug used in the treatment of type II diabetes) reduced epithelial permeability to glucose and prevented the apical growth of S. aureus. Metformin had no direct effect on bacterial growth or on basolateral glucose concentration.

This project will investigate how lung glucose homeostasis is modified by inflammation and hyperglycaemia in vitro and in vivo. It will identify whether metformin inhibits glucose accumulation in ASL and limits the growth of respiratory pathogens in vivo. It will also investigate the therapeutic effect of other drugs which are reported to reduce epithelial permeability or reduce blood glucose concentrations. e.g. azithromycin, glucokinase activators and SGLT2 inhibitors. The outcomes of this project will determine whether these drugs have potential therapeutic benefit in the prevention and treatment of respiratory infection and chronic pulmonary disease.

Planned Impact

This project has potential to impact the public sector, commercial private sector and the wider public. We have identified a drug that has the potential to treat epithelial disease and reduce infection that is distinct from current antibiotic and steroid therapies therefore opening up new possibilities for academic and pharmaceutical research in the short term. Investigation of how this drug works in both in vitro and in vivo systems will add to understanding of the function of epithelial tissues and aid identification of potential new targets for drug development for respiratory disease. This project will therefore directly impact scientists working in epithelia, inflammatory disease, infection and immunity and diabetes.
The main output of this study will be new insights into glucose homeostasis in airway epithelium and its role in preventing infection. As part of this study we will characterise the effects of hyperglycaemia on glucose homeostasis in in vitro and in vivo models of airway epithelial disease. We will use bacterial co-culture/infection techniques to investigate the effect of hyperglycaemia on the growth of respiratory pathogens. The in vitro model we have developed will provide a useful tool for initial drug screening in development of non-antibiotic drugs to prevent lung infection and meets the principles of the 3Rs (Replacement, Refinement and Reduction). We are working with Astra Zeneca to utilize their models of inflammatory lung disease to translate our in vitro findings. They will also provide drugs have been developed but not yet fully available to test in these systems. Their involvement will promote development and translation and therefore enhance impact in this area.
In the long term, we anticipate that understanding of the normal mechanisms that regulate glucose concentrations in the healthy airway, how factors associated with chronic disease and diabetes (prevalent in an ageing population) alter epithelial function and glucose homeostasis, together with the identification of drugs that prevent these effects, will ultimately benefit human and animal health with social and economic impact. Metformin is a drug already in clinical use with very few side effects. Understanding its action on epithelia and how this inhibits glucose flux and the growth of common pathogens such as S. aureus and Ps. aeruginosa provides a new and novel therapeutic mechanism and adds to the known activities of this drug. Data from this study will also add to the known activities of the other drugs studied. We anticipate that this knowledge will facilitate future assessment of the action of these drugs in vivo and translation into treatment could be rapid, with long term benefits to the wider public by enhancing quality of life and health. To enhance impact in this area we already have studies underway to investigate the effect of metformin on exacerbations in patients with chronic obstructive pulmonary disease. Prof. Baker also runs a patient advisory group, comprising patients and their carers who work collaboratively with researchers and clinicians to develop such projects. This group also provides advice for and a route of dissemination of results to patients.
We are currently the only research group in the UK investigating the role of glucose homeostasis in the lung. We are leading the world in this field, which, through publication and presentation of the data at international scientific meetings, is now receiving significant attention overseas. This industrial collaboration will increase impact in this area by aiding translation and development of this novel research into therapies. The success of this project and its future development could therefore contribute to the economic competitiveness of the UK.

Publications

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Baines D (2013) Kinases as targets for ENaC regulation. in Current molecular pharmacology

 
Description Cystic Fibrosis Strategic Research Centre
Amount £718,602 (GBP)
Funding ID SRC 006 
Organisation Cystic Fibrosis Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2016 
End 03/2019
 
Description MRC - London Intercollegiate Doctoral Training Partnership
Amount £105,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2017 
End 03/2021
 
Description MRC CASE studentship
Amount £106,000 (GBP)
Funding ID MR/L013509/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2014 
End 09/2018
 
Description MRC London Intercollegiate Doctoral Training Partnership
Amount £105,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2017 
End 03/2021
 
Description No Scheme
Amount £240,508 (GBP)
Organisation AstraZeneca 
Department Astra Zeneca
Sector Private
Country United States
Start 04/2018 
End 03/2020
 
Title glucose sensors 
Description We have developed several fluorescent glucose biosensors based on 1. Genetically encoded fluorophore linked to protein. 2. Environmental sensitive fluorophore covalently linked to protein. Some of these have now been catalogued for invention disclosure. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? No  
Impact Potential impact for use as a tool to analyse glucose in airway, predict blood glucose, and as a research tool. Helassa N, Garnett JP, Farrant M, Khan F, Pickup JC, Hahn KM, MacNevin CJ, Tarran R, Baines DL. 2014 A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. Biochem J. Sep 15. [Epub ahead of print] PMID: 25220254 
 
Description Astra Zeneca 
Organisation AstraZeneca
Department Respiratory and Inflammation iMED Strategy team Mölndal
Country Sweden 
Sector Private 
PI Contribution Collaborator in MICA award 'Evaluation of anti diabetic drugs in the treatment of respiratory disease'. Collaboration continuing with joint MRC-CASE studentship, and funding for further Astra Zeneca led airway epithelia study, external supervisor for Astra Zeneca Postdoctoral Scientist, Key Opinion Leader.
Collaborator Contribution Collaborator in MICA award 'Evaluation of anti diabetic drugs in the treatment of respiratory disease'. Further funding for Astra Zeneca led project, access to facilities, chemicals and pharmaceutical insight. Joint supervision of MRC-CASE student.
Impact Holmberg CH, Astrand A, Wingren C, Garnett JP, Mayer G, Taylor JD, Baker EH... Baines DL. (2017). Differential Effect of LPS on Glucose, Lactate and Inflammatory Markers in the Lungs of Normal and DiabeticMice. Pulmonary and Respiratory Medicine Open Access 2017. Pulmonary and Respiratory Medicine Open Access 2017.,pp. PROA-101. Åstrand A, Wingren C, Benjamin A, Tregoning JS, Garnett JP, Groves H, Gill S, Orogo-Wenn M, Lundqvist AJ, Walters D, Smith DM, Taylor JD, Baker EH, Baines DL. Dapagliflozin-lowered blood glucose reduces respiratory P. aeruginosa infection in diabetic mice. Br J Pharmacol. 2017 May;174(9):836-847. doi: 10.1111/bph.13741. Epub 2017 Mar 9.
Start Year 2013
 
Description Boehringer Ingelheim Pharma 
Organisation Boehringer Ingelheim
Country Germany 
Sector Private 
PI Contribution Novel data relating to airway epithelial research .
Collaborator Contribution Access to technical expertise/equipment not available at home institution data from which contributed to publication.
Impact doi: 10.1038/srep37955
Start Year 2015
 
Description Chapel Hill Pharmacology 
Organisation University of North Carolina at Chapel Hill
Department UNC School of Medicine
Country United States 
Sector Academic/University 
PI Contribution Development of fluorescent glucose sensor
Collaborator Contribution Provision of long wavelength fluorescent dyes
Impact Helassa N, Garnett JP, Farrant M, Khan F, Pickup JC, Hahn KM, MacNevin CJ, Tarran R, Baines DL. 2014 A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. Biochem J. Sep 15. [Epub ahead of print] PMID: 25220254 Hyperglycaemia and Pseudomonas aeruginosa acidify cystic fibrosis airway surface liquid by elevating epithelial monocarboxylate transporter 2 dependent lactate-H+ secretion. Garnett JP, Kalsi KK, Sobotta M, Bearham J, Carr G, Powell J, Brodlie M, Ward C, Tarran R, Baines DL. Sci Rep. 2016 Nov 29;6:37955. doi: 10.1038/srep37955. PMID:27897253
Start Year 2013
 
Description Francis Crick Institute - Airway basal cells 
Organisation Francis Crick Institute
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution This is a strategic research centre funded by the Cystic Fibrosis trust which involves a partnership between researcher at the Frances Crick Institute. We carry out and provide data from functional experiments to support phenotype replication.
Collaborator Contribution Under the grant funding they provide human primary airway basal cells and culture optimisation for functional experiments.
Impact No outputs directly associated with this collaboration yet. Covers two disciplines cell biology, physiology.
Start Year 2017
 
Description GLUT10 KO mice 
Organisation University of Ghent
Department Department of Pediatrics and Medical Genetics
Country Belgium 
Sector Academic/University 
PI Contribution We are providing data on the effect of GLUT10 KO on the pathophysiology of the airway. This will contribute to our understanding of the function of GLUT10 as a glucose transporter and its role in arterial tortuosity syndrome.
Collaborator Contribution The provision of GLUT10 KO mice for the analysis of airway surface liquid, isolation of airway cells and tissue.
Impact Currently in process of sample collection, data acquisition and analysis.
Start Year 2015
 
Description Glucose transporter knockout mice 
Organisation Technical University of Munich
Country Germany 
Sector Academic/University 
PI Contribution 1. We are providing information on how GLUT 2/5 and SGLT1 KO in mice affects the pathophysiology of the airway. 2. We are providing samples that we collect from the airway to a potential study investigating how glucose transport affects the formation of metabolites in these animals.
Collaborator Contribution The partners are contributing the KO mice to the study for sample and tissue collection.
Impact Currently in the process of data collection and analysis for publication.
Start Year 2015
 
Description Newcastle university 
Organisation Newcastle University
Department Institute of Health and Society
Country United Kingdom 
Sector Academic/University 
PI Contribution Expertise in airway epithelial physiology
Collaborator Contribution Expertise in pH measurement
Impact doi: 10.1038/srep37955
Start Year 2016
 
Description UCL 
Organisation University College London
Department Institute of Child Health
Country United Kingdom 
Sector Academic/University 
PI Contribution Expertise in ion transport measurement across airway cells
Collaborator Contribution Provision of primary airway epithelial cells (HBE) and transformed (HBE)
Impact Sucessful Cystic Fibrosis SRC bid and funding 2016 https://www.cysticfibrosis.org.uk/the-work-we-do/research/research-we-are-funding/strategic-research-centres/src-8-hart-gene-editing. Munye MM, Shoemark A, Hirst RA, Delhove JM, Sharp TV, McKay TR, O'Callaghan C, Baines DL, Howe SJ, Hart SJ (2016) BMI-1 extends proliferative potential of human bronchial epithelial cells whilst retaining their mucociliary differentiation capacity. Am J Physiol Lung Cell Mol Physiol. 2017 Feb 1;312(2):L258-L267. doi: 10.1152/ajplung.00471.2016.
Start Year 2016
 
Description UCL 
Organisation University College London
Department Institute of Child Health
Country United Kingdom 
Sector Academic/University 
PI Contribution Expertise in ion transport measurement across airway cells
Collaborator Contribution Provision of primary airway epithelial cells (HBE) and transformed (HBE)
Impact Sucessful Cystic Fibrosis SRC bid and funding 2016 https://www.cysticfibrosis.org.uk/the-work-we-do/research/research-we-are-funding/strategic-research-centres/src-8-hart-gene-editing. Munye MM, Shoemark A, Hirst RA, Delhove JM, Sharp TV, McKay TR, O'Callaghan C, Baines DL, Howe SJ, Hart SJ (2016) BMI-1 extends proliferative potential of human bronchial epithelial cells whilst retaining their mucociliary differentiation capacity. Am J Physiol Lung Cell Mol Physiol. 2017 Feb 1;312(2):L258-L267. doi: 10.1152/ajplung.00471.2016.
Start Year 2016
 
Description UNC CF and Smoking centre 
Organisation University of North Carolina at Chapel Hill
Department UNC School of Medicine
Country United States 
Sector Academic/University 
PI Contribution How hyperglycaemia contributes to CF disease and effects of smoking on airway. Development of glucose sensor for use in vitro.
Collaborator Contribution provision of primary cultures lung epithelial cells
Impact Helassa N, Garnett JP, Farrant M, Khan F, Pickup JC, Hahn KM, MacNevin CJ, Tarran R, Baines DL. 2014 A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. Biochem J. Sep 15. [Epub ahead of print] PMID: 25220254 Garnett JP, Tarran R, Gray MA, Ward C, Brodlie M, Baker EH and Baines DL (2013). Elevated glucose flux across cystic fibrosis airway epithelium is an important factor for Pseudomonas aeruginosa growth in airway epithelial co-culture. PLoS ONE 8(10): e76283. doi:10.1371/journal.pone.0076283
Start Year 2013
 
Description University of Cork - gene editing 
Organisation University College Cork
Country Ireland 
Sector Academic/University 
PI Contribution We are responsible for functional assessment of gene edited airway epithelial cells
Collaborator Contribution They have provided tools and reagents and expertise to genetically edit cells using CRISPR Cas9 technology.
Impact No outcomes yet
Start Year 2017
 
Title Metformin treatment in COPD 
Description A randomised, double-blind, placebo-controlled trial of metformin in chronic obstructive pulmonary disease (COPD) exacerbations: a pilot study 
Type Therapeutic Intervention - Drug
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2015
Development Status Closed
Clinical Trial? Yes
UKCRN/ISCTN Identifier Sponsor's R&D Registration Number: 10.0086
Impact This intervention strategy with metformin did not ameliorate elevations in blood glucose concentration among non-diabetic patients admitted to hospital for COPD exacerbations, and had no detectable effect on CRP or clinical outcomes. 
 
Description Princes Teaching Institute 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Invitation to present a talk discussion on Transporters to a New Teacher Subject Day, Westminster. The purpose of the day was to provide up to date scientific information and insight to help teachers translate into classroom and enthuse students. This is now an ongoing appointment
Year(s) Of Engagement Activity 2015
URL http://www.princes-ti.org.uk/what-we-do/new-teacher-subject-days
 
Description Public Engagement Event 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact The event was Living with Type 1 Diabetes

It sparked a lot of discussion and increased awareness about research and patient experience

There were a lot of interesting question from staff and public. The event sparked public interest in future events
Year(s) Of Engagement Activity 2014
URL http://www.sgul.ac.uk/news/public-events
 
Description Schools visits (London) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Schools
Results and Impact 30 or more 15-18 year students attend talks on CF disease (AS/A2 module subject) or research. Ususally sparks discussion and leads to work experience placements at SGUL.

Sparks discussion about research and entrance to biomedical/medical degree courses. Regularly leads to work experience placements at SGUL.
Year(s) Of Engagement Activity Pre-2006,2006,2007,2008,
 
Description U3A 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 70 people attended the lecture, good discussion and questions after the event

request for further speakers
Year(s) Of Engagement Activity 2014