A study of ciliary function and inflammation in respiratory tract epithelium

Motile cilia on epithelial cells in the airways, nose, paranasal sinuses, and middle ear beat over 1 million/day clearing mucus, inflammatory mediators, pathogens and pollutants. A number of diseases cause abnormal ciliary function and these diseases are also associated with airway inflammation, for example viral infection, bacterial infection, severe asthma, chronic obstructive pulmonary disease and primary ciliary dyskinesia. The group has extensive experience in ciliary biology in disease and the collaboration between UCL and AstraZeneca will allow detailed exploration of the role of ciliary function in inflammation.
Under the hypothesis that normal ciliary function is anti-inflammatory, this studentship aims to identify:
1. Mechanisms by which ciliary stasis or dyskinesia are associated with inflammation.
2. Whether mechanical perturbation in an attempt to replicate cellular sheer stress from ciliary beating abrogates inflammation.
The applicants have created an extensive biobank of epithelial tissue that can be cultured at an air liquid interface allowing differentiation into ciliated epithelium using co-culture methods developed by the applicants. Multiple epithelial samples are available from healthy individuals, severe asthmatics, and also from individuals with specific gene defects that produce static cilia (DNAH5 n = 8) and cilia with a normal beat frequency but markedly reduced ciliary amplitude (CCDC40 n = 6). The student will culture these cells to a ciliated phenotype.
Readouts to assess inflammatory parameters will include ciliary beat frequency and beat pattern (high speed video microscopy); particulate movement; inflammatory mediator release (cytokines/chemokines); NO; reactive oxygen species. Isolation of ciliated cells using flow cytometry using methods developed by the applicants will allow RNAseq analysis.
Readouts will allow investigation of epithelium with: normal cilia, static cilia, cilia with normal frequency but markedly reduced amplitude and dysfunctional cilia due to increased viscous and asthma to determine the association between dysfunctional cilia and enhance inflammation.