Dr
Diane
Lee
Diane is an ex vivo and in vitro cell biologist with considerable experience in drug discovery and pre-clinical research in both industry and academia.
Having embarked upon a career in oncology and cancer therapeutics, Diane spent time at the Institute of Cancer Research, developing cell based and biochemical assays. From here she moved into the field of epigenetics at the Marie Curie Research Institute, studying the role of TAF5 in eukaryotic gene transcription, using site-directed mutagenesis and working with colleagues to develop an in vitro model of eukaryotic gene transcription. Her work was crucial in the confirmation of the dimeric structure of this transcription factor. Diane went on to gain extensive technical expertise in primary and 3D tissue culture at Novartis, working in the Gastro-Intestinal Disease Area in Horsham. Here she developed culture techniques of human and mouse small intestinal epithelial cells (SIECs) and colonic epithelial cells (CECs) as organoids, using them to study mechanisms of mucositis and therapeutics/prevention thereof. Upon the closure of GIDA, she moved back into academia, starting her PhD in 2011 at the School of Pharmacy and Biomolecular Sciences at the University of Brighton. Here she developed an in vitro model to enable pre-clinical identification of drug irritancy and permeability issues in the lung as part of ADME determination, completing her PhD in 2015.
Currently, Diane fulfils the role of Research Fellow B in Pathology and Infectious Diseases and is a regular participant and instigator of collaborations (internal and external), working with colleagues at NIBSC, Pirbright and APHA. Her current research interests are in the development an equine nasal brush model, with the aim of utilising the model to study equine asthma and the role of Notch signalling in goblet cell hyperplasia. She is passionate about championing the principles of the 3Rs; Replacement, Refinement and Reduction of animals in scientific experiments.
Equine asthma, formerly inflammatory airway disease (IAD), is amongst the most common causes of training interruption and poor performance in young athletic horses, particularly the racing thoroughbred and endurance horse. The prevalence is high in racehorses (13–22%) and sports horses (31%) and may be described as a chronic respiratory syndrome affecting horses of any age, gender or breed. Research on asthma is hindered by the invasiveness of the techniques required to isolate tracheal or bronchial epithelial cells. Meaningful quantities of upper airway epithelial cells can currently only be achieved post-slaughter, whilst bronchial brushing or biopsy require local anaesthesia in addition to sedation and require a 48 h recovery period.
The current project, funded by the Horserace Betting and Levy Board (HBLB Grant reference 793), aims to develop a model of the equine upper airway, using a nasal brush sampling technique, as an alternative to the more invasive tracheo-bronchial biopsy or bronchial brushing. Nasal epithelial cells have been validated as a surrogate for bronchial epithelial cells in human cystic fibrosis studies. Nasal epithelial cells from healthy horses will be characterised, validated by comparison with cells isolated from the equine bronchi and used to study mechanisms of mild and severe equine asthma. We will also investigate modulation of the notch signalling pathway by gamma-secretase inhibitors. Notch signalling is responsible for the differentiation of basal and ciliated cells into goblet cells, which leads to excess mucus and poor performance in asthma.
Previously, my research was centred around a strategic grant (NC/M002047/1), awarded by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), entitled “A bovine alveolus model to replace cattle in the study of host-pathogen interactions in bovine tuberculosis.”
BTB is a zoonosis which infects livestock and wildlife with severe socio-economic consequences and an impact on animal health. In the absence of improved control the projected economic burden to GB over the next decade is predicted to be £1 billion (http://www.defra.gov.uk/animal-diseases/a-z/bovine-tb/). Tackling BTB requires deeper insights into host-pathogen interactions otherwise it is unlikely any major breakthroughs in developing effective tools for disease intervention will occur. We developed a tissue culture model with which to study fundamental events following infection of the bovine lung with virulent mycobacteria that can’t be conducted currently in vitro. Our model consists of a cellular co-culture of bovine pulmonary artery endothelial cells (BPAECs) and bovine alveolar type II (immortalised and primary) cultured at an air-liquid interface on a PET porous membrane. We are using this model to test the hypothesis that a significant aspect of vaccine-mediated protection against BTB is expressed at the level of host-pathogen interactions within the alveolus.