Ivanova, RF; (2017) Developing a new nanoprobe research toolbox for studying airway epithelia with applications in cystic fibrosis. Doctoral thesis , UCL (University College London).

Abstract

The airways are lined with a thin layer of fluid called the airway surface liquid (ASL), whose function is to protect the lungs from mechanical damage and infection. It is a complex fluid with tightly regulated properties which are crucial for pulmonary health. In cystic fibrosis (CF), some of these properties are disrupted. CF is caused by loss-of-function mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel. This has vast downstream effects on airway epithelial function and ASL properties. In order to study these effects, a novel environmental chamber was constructed, where samples were maintained at physiological conditions. The chamber was fitted onto the stage of a scanning ion conductance microscope (SICM). SICM is a scanning probe super-resolution imaging technology which was, as described in this thesis, developed into a comprehensive research toolbox for airway epithelial studies. The toolbox included methods for measuring transepithelial potential, ASL depth, ASL pH, ASL viscoelasticity, cilia availability and beating. These methods were used to compare bronchial epithelial cultures derived from healthy and CF donors. It was found that the CF samples were dehydrated compared to controls, in agreement with many previous CF studies. Rehydration of CF cultures was attempted by reducing apical Na+ absorption through mRNA silencing, and by application of the CF treatment Orkambi. In addition, the role of K+ transport in regulating ASL depth was also investigated. K+ channel modulation was identified as a potential therapeutic approach to improve the hydration of CF airway epithelia. In summary, a powerful new approach to measuring epithelial/ASL function was developed to create a toolkit for ASL and epithelial cell studies. The capability of this toolkit was illustrated in studies of CF epithelia. However, it is not limited to the study of CF, and could well find importance in other studies of the respiratory airways, particularly in diseases such as asthma and chronic obstructive pulmonary disease, where the ASL is also disrupted.

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