Primary bronchial epithelial cell culture from people with cystic fibrosis (CF) - a model to study CF lung disease
Lead Research Organisation:
Newcastle University
Department Name: Clinical Medical Sciences
Abstract
Over 7000 people have cystic fibrosis (CF) in the United Kingdom making it the most common life-limiting inherited disease. Death and illness in CF are largely due to progressive lung disease. The disease is characterised by severe inflammation, particularly involving neutrophilic pus cells, and structural airway damage.
Neutrophilic inflammation is one of the earliest findings in CF occurring in young children prior to obvious infection. Interleukin(IL)-17, a chemical mediator produced by the body, has attracted great interest recently as a driver of neutrophil-pus cells in a range of inflammatory diseases.
In CF there is evidence that concentrations of IL-17 rise in sputum when patients become unwell.
We are the first group in Europe to establish the invaluable experimental resource of culturing cells from the lungs of actual people with CF at the time of transplantation.
The first studies using our new model of CF lung disease will focus on the role of IL-17. An improved understanding of the inflammatory process may lead to new treatments capable of alleviating the inflammation and lung damage that is such a problem in CF.
Neutrophilic inflammation is one of the earliest findings in CF occurring in young children prior to obvious infection. Interleukin(IL)-17, a chemical mediator produced by the body, has attracted great interest recently as a driver of neutrophil-pus cells in a range of inflammatory diseases.
In CF there is evidence that concentrations of IL-17 rise in sputum when patients become unwell.
We are the first group in Europe to establish the invaluable experimental resource of culturing cells from the lungs of actual people with CF at the time of transplantation.
The first studies using our new model of CF lung disease will focus on the role of IL-17. An improved understanding of the inflammatory process may lead to new treatments capable of alleviating the inflammation and lung damage that is such a problem in CF.
Technical Summary
Backgound
Over 7000 people in the United Kingdom have cystic fibrosis (CF) making it the most common life-limiting genetic disorder. Although life-expectancy has increased with improvements in clinical care there remains an increasing burden of morbidity due to progressive lung disease. The CF airway is characterised by intense neutrophilic inflammation, even prior to infection. Chronic disease is also associated with mucus gland hyperplasia.
As part of the lung transplantation programme in Newcastle I have successfully established a technique of culturing primary bronchial epithelial cells (PBECs) from the explanted lungs of people who have CF.
Interleukin-(IL)17 has recently been identified as a major orchestrator of neutrophilic inflammation and plays an important role in several inflammatory conditions. Animal studies have demonstrated that administration of IL-17 to airways results in neutrophil accumulation. IL-17 also increases expression of the mucin genes MUC5AC and MUC5B by bronchial epithelial cells.
In summary there is evidence from animal and cellular studies that IL-17 is linked to neutrophilic inflammation and mucus excess that are the cardinal features of the CF phenotype. In addition, people with CF have increased levels of IL-17 in sputum during infective exacerbations. Concentrations of IL-23, a cytokine that regulates IL-17, are similarly raised.
Aims and Objectives
I aim to utilise my patient-derived PBEC model to investigate the hypothesis that IL-17 is associated with the neutrophilic inflammation and mucus excess characteristic of CF lung disease.
Design and Methodology
1. Histology: I will assess airway architecture and inflammation in explanted CF lungs compared to controls by tinctorial stains (collagen and mucus) and immunohistochemistry (IL-17,CD4,CD8,neutrophil elastase)
2. PBEC Culture: I will establish cultures from explanted CF lungs. Submerged cultures will be transferred to air-liquid interface to promote differentiation and mucus production. Control cultures will be established via bronchoscopic brushings from healthy individuals.
3. Inflammation and Mucus: I will assess the response of early passage CF and control PBECs to IL-17 stimulation. Neutrophilic cytokines (multiplex ELISA), mucus (ELISA) and mucin gene expression (PCR) will be measured.
Scientific and Medical Opportunities
Primary cells are seldom available for CF research. At the largest UK lung transplant centre I can establish a unique experimental resource of culturing PBECs from CF lungs. This will provide a model to study the pathway of inflammation and other potential therapeutic targets. This project will provide me with a superb training opportunity to increase my skills in clinical research, obtain a PhD
Over 7000 people in the United Kingdom have cystic fibrosis (CF) making it the most common life-limiting genetic disorder. Although life-expectancy has increased with improvements in clinical care there remains an increasing burden of morbidity due to progressive lung disease. The CF airway is characterised by intense neutrophilic inflammation, even prior to infection. Chronic disease is also associated with mucus gland hyperplasia.
As part of the lung transplantation programme in Newcastle I have successfully established a technique of culturing primary bronchial epithelial cells (PBECs) from the explanted lungs of people who have CF.
Interleukin-(IL)17 has recently been identified as a major orchestrator of neutrophilic inflammation and plays an important role in several inflammatory conditions. Animal studies have demonstrated that administration of IL-17 to airways results in neutrophil accumulation. IL-17 also increases expression of the mucin genes MUC5AC and MUC5B by bronchial epithelial cells.
In summary there is evidence from animal and cellular studies that IL-17 is linked to neutrophilic inflammation and mucus excess that are the cardinal features of the CF phenotype. In addition, people with CF have increased levels of IL-17 in sputum during infective exacerbations. Concentrations of IL-23, a cytokine that regulates IL-17, are similarly raised.
Aims and Objectives
I aim to utilise my patient-derived PBEC model to investigate the hypothesis that IL-17 is associated with the neutrophilic inflammation and mucus excess characteristic of CF lung disease.
Design and Methodology
1. Histology: I will assess airway architecture and inflammation in explanted CF lungs compared to controls by tinctorial stains (collagen and mucus) and immunohistochemistry (IL-17,CD4,CD8,neutrophil elastase)
2. PBEC Culture: I will establish cultures from explanted CF lungs. Submerged cultures will be transferred to air-liquid interface to promote differentiation and mucus production. Control cultures will be established via bronchoscopic brushings from healthy individuals.
3. Inflammation and Mucus: I will assess the response of early passage CF and control PBECs to IL-17 stimulation. Neutrophilic cytokines (multiplex ELISA), mucus (ELISA) and mucin gene expression (PCR) will be measured.
Scientific and Medical Opportunities
Primary cells are seldom available for CF research. At the largest UK lung transplant centre I can establish a unique experimental resource of culturing PBECs from CF lungs. This will provide a model to study the pathway of inflammation and other potential therapeutic targets. This project will provide me with a superb training opportunity to increase my skills in clinical research, obtain a PhD
