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.
Areas of specialism
University roles and responsibilities
- Athena Swan SAT member (Vet School)
- Research Culture Committee representative
- NASPA Sub-committee member
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), aims to develop a model of the equine upper airway, using a nasal brush sampling technique, as an alternative to the more invasive tracheobronchial 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 gammasecretase 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 aim to replace cattle in the study of BTB pathogenesis by providing 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 bilayer 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.
This project has a positive impact on animal replacement, removing the need to infect cattle with mycobacteria to answer fundamental questions in TB pathogenesis and provide a valid substitute for cattle that can be used by researchers without access to animal facilities. Vaccines against BTB developed to generate a specific host response would be a significant advance on the current situation where vaccines are tested empirically in cattle to evaluate their efficacy and identify biomarkers that might predict vaccine efficacy without challenge. A specific objective of this project is to determine whether the behaviour of BCG / M. bovis and host cells in the model correlates with the protective efficacy seen in cattle challenge studies from which we have stored PBMCs to evaluate.
- VMS1003 – Structure and Function 1
The molecular basis of genetics I - Nucleic acids and chromatin
The molecular basis of genetics II – DNA Replication
Gene Expression I and II - Transcription and Translation
- Supervision and direction of undergraduates and visiting researchers in laboratory projects.
Alveolar type II (ATII) cells play a key role as part of the distal lung epithelium, including in the innate immune response and as self‐renewing progenitors to replace alveolar type I (ATI) cells during epithelial regeneration. Their secretion of surfactant protein helps maintain homeostasis and exerts protective, antimicrobial properties. ATII cells remain difficult to study, partly due to inefficient and expensive isolation methods, a propensity to differentiate into ATI cells, and susceptibility to fibroblast contamination. Published methods of isolation often require specialized technology, negatively impacting the development of in vitro models of disease, including bovine tuberculosis. Presented here is a simple and cost‐effective method for generation of bovine primary ATII cells. These cells exhibit an ATII phenotype in 2D and 3D culture and are conducive to further study of the role of ATII cells in bovine respiratory diseases.
Alveolar type II (ATII) cells play a key role as part of the distal lung epithelium, including roles in the innate immune response and as self-renewing progenitors to replace alveolar type I (ATI) cells during regeneration of the alveolar epithelium. Their secretion of surfactant protein helps to maintain homeostasis in the distal lung and exert protective, antimicrobial properties. Despite the cell’s crucial roles, they remain difficult to study, in part due to inefficient and expensive isolation methods, a propensity to differentiate into alveolar type I cells in culture and susceptibility to fibroblast overgrowth from primary isolations. Published methods of isolation often require specialist technology, negatively impacting the development of in vitro models of disease, including bovine tuberculosis (BTB), a serious re-emerging disease in both animals and humans worldwide. We present here a simple and cost effective method that may be utilised in the generation of bovine primary ATII cells. These exhibit an ATII phenotype in 2D and 3D culture in our studies and are conducive to further study of the role of ATII cells in bovine respiratory diseases.
Background: Aedes aegypti is a vector for the (re-)emerging human pathogens dengue, chikungunya, yellow fever and Zika viruses. Almost half of the Ae. aegypti genome is comprised of transposable elements (TEs). Transposons have been linked to diverse cellular processes, including the establishment of viral persistence in insects, an essential step in the transmission of vector-borne viruses. However, up until now it has not been possible to study the overall proteome derived from an organism’s mobile genetic elements, partly due to the highly divergent nature of TEs. Furthermore, as for many non-model organisms, incomplete genome annotation has hampered proteomic studies on Ae. aegypti. Results: We analysed the Ae. aegypti proteome using our new proteomics informed by transcriptomics (PIT) technique, which bypasses the need for genome annotation by identifying proteins through matched transcriptomic (rather than genomic) data. Our data vastly increase the number of experimentally confirmed Ae. aegypti proteins. The PIT analysis also identified hotspots of incomplete genome annotation, and showed that poor sequence and assembly quality do not explain all annotation gaps. Finally, in a proof-of principle study, we developed criteria for the characterisation of proteomically active TEs. Protein expression did not correlate with a TE’s genomic abundance at different levels of classification. Most notably, long terminal repeat (LTR) retrotransposons were markedly enriched compared to other elements. PIT was superior to ‘conventional’ proteomic approaches in both our transposon and genome annotation analyses. Conclusions: We present the first proteomic characterisation of an organism’s repertoire of mobile genetic elements, which will open new avenues of research into the function of transposon proteins in health and disease. Furthermore, our study provides a proof-of concept that PIT can be used to evaluate a genome’s annotation to guide annotation efforts which has the potential to improve the efficiency of annotation projects in non-model organisms. PIT therefore represents a valuable new tool to study the biology of the important vector species Ae. aegypti, including its role in transmitting emerging viruses of global public health concern.
Lee, D.F. and Chambers, M.A. A bilayer model of the bovine alveolus to study bovine respiratory disease and host-pathogen interactions. F1000Research, 2019. DOI: 10.12688/f1000research.18696.1
Lee, D.F. and Chambers, M.A. Isolation of Alveolar Type II (ATII) cells from adult bovine lung. Current Protocols in Toxicology 2019. DOI: 10.1002/cptx.71
Lee, D.F., Salguero-Bodes, F.J., Grainger, D., Francis, R.J., MacLellan-Gibson, K., Chambers, M.A. Isolation and characterisation of alveolar type II pneumocytes from adult bovine lung. Sci Rep 2018 DOI: 10.1038/s41598-018-30234.
Jones, A.M., Westwood, I.M., Osborne, J.D., Lee, D.F., McAndrew C., Jones K., Workman P., Collins I., van Montfort R.L. A fragment-based approach applied to a highly flexible target: Insights and challenges towards the inhibition of HSP70 isoforms. Sci Rep. 2016 Oct 6;6:34701. doi: 10.1038/srep34701.
Malecová, B., Caputo, V.S., Lee, D.F., Hsieh, J.J., Oelgeschläger T. Taspase1 processing alters TFIIA cofactor properties in the regulation of TFIID. Transcription. 2015;6(2):21-32. doi: 10.1080/21541264.2015.1052178.
Lee DF, Salguero FJ, Stewart GR, Villareal-Ramos B, Vordermeier M, Chambers, MA: A bilayer model of BCG interaction with the bovine alveolus. Advances in Cell and Tissue Culture, Cardiff, 2019
Lee DF, Salguero FJ, Stewart GR, Vordermeier M, Chambers, MA: The use of an immortalised bovine alveolar type II (B2AE/BATII) cell line in the construction of a bilayer model of the alveolus. NC3Rs Fellows Meeting, London, 2018
Lee DF, Chambers MA: Construction and characterisation of a bilayer consisting of bovine type II alveolar epithelial (BATII) and bovine pulmonary arterial endothelial cells (BPAECs). Advances in Cell and Tissue Culture, Cardiff, 2018.
Lee DF, Chambers MA: Immortalisation and characterisation of a bovine type II alveolar epithelial (B2AE) cell line. Association for Veterinary Teaching and Research Work annual conference, 2017
Lee DF, Lethem MI and Lansley AB: A mucus-secreting in vitro cell culture model (SPOC1) for studying drug absorption [of beta-blockers] in the airways, PharmSci, Hatfield 2017
Lee DF, Salguero-Bodes FJ, Stewart GR, Vordermeier M, Chambers MA: A bovine alveolus model to replace cattle in the study of host-pathogen interactions in bovine tuberculosis (BTB). Advances in Cell and Tissue Culture Research, Manchester, 2017
Lee DF, Salguero-Bodes FJ, Stewart GR, Vordermeier M, Chambers MA: A bovine alveolus model to replace cattle in the study of host-pathogen interactions in bovine tuberculosis (BTB). Vaccinology, Belfast, 2017 Invited Speaker
Lee DF, Salguero-Bodes FJ, Stewart GR, Vordermeier M, Chambers MA: A bovine alveolus model to replace cattle in the study of host-pathogen interactions in bovine tuberculosis (BTB). NC3Rs Symposium, 2016
Smrekar K, Fox R, Lee D, Gosling M and Danahay H: Flagellin-induces a hypersecretory phenotype in primary human bronchial epithelial cells, presented at The 13th European Cystic Fibrosis Basic Science Conference, Pisa, Italy, 2016
Lansley AB, Lee DF and Lethem MI: Are respiratory cell lines proving useful in pharmaceutical development? Drug Delivery to the Lungs Conference (DDL25), The Aerosol Society, Edinburgh 2014.
Lee DF, Lansley AB, Lethem MI: The characterisation of the UNCN3T airway cell line as an in vitro model to study drug permeability in the presence of mucus (presentation and poster). PharmSci 2015, Nottingham; British Association of Lung Research, 2014, London; Doctoral College Conference, University of Brighton, 2014.
Lee DF, Lansley AB, Lethem MI: SPOC1: a secretagogue-free in vitro model to study drug permeability in the presence of mucus (presentation and poster). PharmSci 2014, Hatfield; British Association of Lung Research, 2014, London; Doctoral College Conference, University of Brighton, 2014.
Lee DF, Lansley AB, Lethem MI: The use of SPOC1 as an in vitro model to study drug permeability in the presence of mucus (presentation, poster and position on the discussion panel). PharmSci 2013, Edinburgh; Doctoral College Conference, University of Brighton, 2013.
Lee DF, Lansley AB, Lethem MI: The use of Calu-3 as an in vitro model to study drug permeability in the presence of mucus (presentation and poster – manuscript in preparation). Doctoral College Conference 2012, University of Brighton; British Association of Lung Research, 2012, Southampton, 2012.
Lee DF, Lansley AB, Lethem MI: The use of in vitro models to study drug permeability in the presence of mucus (presentation). GlaxoSmithKline Lung Symposium, Stevenage (video-link to US), 2011.
Lee DF, Oelgeschlager T: The Functional Characterisation of hsTAF5 (poster and presentation). Marie Curie Research Institute Conference, Wye, 2007.
Lee DF, Aherne, GW: The HTS of upregulation of the mis-match repair (MMR) gene MLH1 in HCT116 (poster). British Association of Cancer Research, Leeds, 2002