Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.778491
Title: Designing and utilising an in vitro model to investigate cellular pathways involved in idiopathic pulmonary fibrosis and connective tissue disease associated interstitial lung disease
Author: Cotton, Caroline Victoria
ISNI:       0000 0004 7964 2224
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2019
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Abstract:
ILD represents a heterogeneous spectrum of diseases caused by excessive deposition of extracellular matrix (ECM) into the interstitial space leading to distortion of the lung architecture and impairment of gas exchange. In this thesis the design of an in vitro model is described to test the pathways involved in the fibrotic response. The model was first tested on fibroblast and epithelial cell lines, two cell types that are implicated as key players in pulmonary fibrosis, treating cells with the pro-inflammatory cytokine Transforming Growth Factor-β (TGF-β), ensuring that this model was reliable and reproducible. Once a model had been established, it was utilised by treating cells with plasma from patients with IPF and Systemic Sclerosis Associated ILD (SSc-ILD). This was to assess whether treatment with plasma from patients with fibrotic lung disease would induce any changes in these cell lines. When cells were treated with plasma there were no changes in gene expression of the MRC5 fibroblasts but there was an increase in proliferation of the fibroblast cells when treated with IPF patient plasma. When the A549 epithelial cells were treated with plasma from patients with IPF, there was a significant reduction in COL3A1 and FN-EDA. This was associated with a significant downregulation of N-cadherin. These changes are not those expected in EMT. Conversely treatment of the same cell line with SSc-ILD plasma led to a significant increase in gene expression of COL3A1 and FN-EDA. Finally, this model was adapted to imitate the constant movement of the lungs by stretching the cells in vitro. Stretch alone caused no significant differences in any of the genes associated with fibrosis in either cell type. However, when used alongside TGF-β treatment there was an attenuation of the fibrotic response in COL1A1, FN-EDA and CCN2 in the fibroblasts and COL1A1 and Slug in the epithelial cells. In conclusion, this thesis has produced a novel way to explore some of the key pathways involved in fibrosis and the changes that plasma treatment induces on cell lines.
Supervisor: Cooper, R. G. ; Bou-Gharios, G. ; Spencer, L. G. ; Lamb, J. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.778491  DOI:
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