Activities of the Team
Jane Alder has developed a novel human three-dimensional in vitro blood brain barrier (BBB) glioma model from primary cells with realistic architecture and dynamic flow optimised for functional analysis of metabolic and transporter proteins. The aim of her research is to further understanding of molecular mechanisms involved in maintaining BBB integrity and function during gliomagenesis and metastasis, as well as modelling drug disposition to the CNS for in vivo pharmacokinetic predictions. Current projects have focused on: 1) microRNA-epigenetic regulatory circuit of transporter expression; 2) use of aptamer targeting ligands for transport at the BBB; 3) the role of folate metabolism, transport and epigenetics in programming of gliomagenesis and iv) the exploitation of impaired bioenergetic states and mitochondrial mutations in glioma for targeted therapy with isolated dietary lipids.
Dr Lisa Shaw has discovered a glioblastoma specific miRNA signature released from exosomes into the circulation during gliomagenesis. This was the first report of miRNA expression based on the age and sex of glioma patients. Current projects are investigating the measurement of miRNA expression in CSF and serum biofluids as a non-invasive diagnostic test and for prediction of response to therapy and outcome. Lisa has also identified a novel target ku70/80 differentially expressed on glioma cells and recognised by the aptamer SA43. The ultimate aim is to create a therapeutically active aptamer-microRNA hybrid that can selectively target glioma cells as well as restoring dysregulated microRNA expression. In addition the role of histone deacetylase for epigenetic regulation and modulation by inhibitors has been investigated. Lisa has established a pre-clinical xenograft model of glioma and is presently investigating the efficacy of these novel therapeutics in vivo.
Dr Clare Lawrence’s main interest lies firmly in the use of yeast models (budding yeast Saccharomyces cerevisiae, fission yeast Schizosaccharomyces pombe, and oleaginous yeast Lipomyces starkeyi) as analogues for key brain tumour signalling pathways. The MAPK and mTOR pathways and are highly conserved across species, making the yeast model an ideal tool for studying lipid accumulation, often observed in brain tumours. Clare found a novel role for stress activating protein kinase pathways in regulating lipid accumulation in yeast and demonstrated cross talk between the MAPK and TOR pathways suggesting that directly targeting these pathways may reduce tumour associated lipogenesis, and hence glioma growth. Clare’s research is currently focussed on: 1) using aptamers for identification of differentially expressed targets in oligodendroglioma and 2) identifying novels aptamers against the mutated EGFR vIII often expressed in glioblastoma and associated with rapid proliferation. To further this aim, Clare has developed a more stringent method for generating aptamers against yeast and glioma targets through incorporation of multiple, sequential or combined negative selections during SELEX (systematic evolution of ligands by systematic enrichment). In addition she has established a novel aptamer-precipitation method for identifying aptamer bound protein targets.
Dr Chris Smith’s experience of in vivo microdialysis and intracranial surgery is being applied to brain cancer to establish a murine model of glioblastoma at UCLan. This model is being used to test potential treatments developed at UCLan and by external collaborators. In addition we are studying circulating microRNA biomarkers found in the serum from glioma patients and in the murine model. The effect these microRNAs may have on glioma growth is being investigated to demonstrate functional significance of the altered circulating profile. This work may help the glioma to be diagnosed earlier and thus improve prognosis.
Prof Kamalinder Singh Malignant brain cancer treatment is limited by a number of barriers, including the blood–brain barrier, transport within the brain interstitium, difficulties in delivering therapeutics specifically to tumour cells, the highly invasive quality of gliomas and drug resistance. As a result, the prognosis for patients with high-grade gliomas is poor and has improved little in recent years. Nanotechnology has emerged as an exciting and promising new means for enabling CNS negative drugs to cross the BBB, diffuse within the brain tissue, target specific cell or signalling systems, and act as vehicles for delivering therapeutics. Research within our group focuses on development of QbD-enabled robust and scalable hybrid protein nanoparticles and lipid-based nanoparticle platforms that have shown promise for crossing BBB. The nanocarrier systems are being investigated to be surface modified with different ligands including Mabs, peptides, Aptamers and bioactive fatty acids for specific targeting to glioblastoma.
Prof Frank Martin has a long standing interest in genetic toxicology, bio-imaging and biospectroscopy and has published extensively in the field of cancer diagnosis. He found that infrared (IR) and/or Raman spectroscopy combined with multivariate analysis could be applied to discriminate between normal brain tissue and different tumour types (meningioma, glioma and brain metastasis) based on the unique spectral “fingerprints” of their biochemical composition. His research is currently concerned with translating vibrational spectroscopy of biofluids from the laboratory to a clinical setting for disease screening or diagnosis.
Dr Tim Snape has conducted research on a family of 2-arylindoles and Indole-3-carbinol which have been evaluated for their anticancer activity against glioblastoma cell lines using different cell viability assays. Initial studies suggest that their mechanism of action is consistent with the generation of reactive oxygen species followed by autophagic cell death. Additionally, functionalised conformationally restricted N,N’-dimethyl-N,N’-diarylureas, which occupy a similar 3-dimensional space to combretastatin A-4 (CA4), have been evaluated by Dr Snape’s group for their ability to inhibit tubulin polymerisation and inhibit growth of short term GBM cell cultures and the established GBM cell line (U251MG). Results show that the ureas closely resembling CA4, regarding benzene ring oxygenation and overall shape, are the most active, leading to the further development of this series of compounds. Dr Snape also has an interest in foldamers, oligomeric ureas of m-phenylenediamine, which his group has shown to target anionic DMPG (dimyristoylphosphatidylglycerol) and possess promise as membrane disruptive agents. Their similar size, shape and hydrophobicity to helical biologically active peptides may be important for activity and the ability of these compounds to insert into a well ordered lipid environment. These lead compounds will be further investigated and developed as novel anticancer agents.
Dr Gail Welsby’s group has investigated the anticancer properties of a range of natural products including flavonoids (quercetin) and triterpenes (asiatic acid) whilst simulating the hypoxic tumour microenvironment. The therapeutic potential of asiatic acid, was shown to have anti-proliferative, pro-apoptotic and reduced wound healing effects on brain cancer cell lines in vitro under hypoxia. Her future work aims to overcome challenges to crossing of the blood brain barrier (BBB) and maintaining a therapeutic concentration by characterising a nanoparticle drug delivery system (DDS) that will allow for efficient delivery of the herbal extract to the brain.
Dr Philip Welsby is examining the therapeutic potential of aspirin analogues for the treatment of glioma in collaboration with Dr Iain Nicholl and Dr Chris Perry of Wolverhampton University. Over the last 30 years aspirin has been linked to a reduced risk of developing a variety of cancers driving the development of analogues with increased anti-cancer efficacy, however previous studies only focused on the short term efficacy of millimolar aspirin concentrations. Philip has found the lead aspirin analogue showed decreased cell proliferation, decreased population of G0/G1 phase cells in cell cycle analysis, decreased cyclin D1 and EGFR activation, and total EGFR expression. Apoptosis was induced in a concentration and time dependent manner in both the cell lines and short term cultures, with activation of both intrinsic and extrinsic pathways. Finally, reduced migration in both the Boyden chamber and scratch assays was seen, but invasion was not inhibited. In conclusion, his data demonstrated that low micromolar concentrations inhibited cell proliferation and reduced viability in glioma cell lines and supported the further development of the aspirin analogue as a novel therapeutic drug for the treatment of glioma.
Dave Griffiths is a state registered (Health Professionals Council) Senior Biomedical Scientist and had worked for 15 years in the Cellular Pathology Department of Royal Preston Hospital prior to commencing his role of Senior Lecturer at UCLan. Dave’s research interests are focussed around neuropathology and he has supervised and contributed to various projects across the Neuro-oncology Research Group, for example: 1) the development of aptamer histological and cytological staining techniques; 2) investigating the tissue type (normal, cancerous and metastatic) and tissue preparation for spectral histopathology by Raman microspectroscopy and 3) developing methods for miRNA biomarker detection and localisation in patient glioma tissue sections using in situ hybridisation.
Pete Abel is a registered Biomedical Scientist specialising in Haematology and he worked in the NHS for 19 years, before coming to UCLan as a Senior Lecturer. Pete is currently undertaking his PhD which aims to identify a range of cytokines and other growth/differentiation factors in patient serum for diagnosis of both low-grade and high-grade glioma patients and prediction of response to treatment. Using a ‘Bio-Plex®’ analyser, up to 100 separate analytes from one 50 µl serum sample can be measured simultaneously. Results from patients have identified several serum factors with significantly altered concentrations in comparison to age-matched controls. Furthermore, a significant fall in levels is observed following resection and encourages support for a large scale longitudinal study for glioma patients in order to identify potential biomarkers that enable clinicians to predict tumour recurrence.
Dr Julie Burrow is Manager of the Cell Culture Laboratories in Darwin Building where the majority of the work by the Neuro-oncology Research Group is conducted. Julie has full responsibility for the Cell Culture Training Programme run for all new staff and students as well as training on the flow cytometer and confocal microscope. In addition, Julie is a key member of Brain Tumour North-West, where she liaises closely with the management team to guarantee information is passed to the researchers for example by ensuring standardised cell culture protocols are followed to allow for direct comparison of results between external collaborators and HTA regulations are adhered to. At present, Julie is working to establish and characterise a short-term culture cell bank from primary glioma tissue donated via the brain tissue bank.
Dr Sarah Dennison is a Senior Research Fellow and Manager of the Biomedical Research Facility. Her research is concerned with investigating anticancer α-helical peptides and structure / function relationships underpinning interactions with tumour cell membranes. Recently she has found that amphibian and plant anionic host defence peptides (HDPs), kill glioma cell lines via membranolytic mechanisms. The ability of HDPs to partition into glioma membranes was investigated in Langmuir-Blodgett troughs and interactions were modelled using Molecular Dynamic (MD) simulations.
Dr Izabela Stasik recently joined the Neuro-oncology Research Group and has brought with her extensive experience in basic research, investigating impairment of apoptosis and mechanisms of resistance to therapeutics, as well as research on potential molecular targets for novel therapies. Currently Izabela collaborates with Professor Mehmet T. Dorak, who uses Genome-Wide Association Studies (GWAS), for study of genetic markers linked to susceptibility to a specific type of cancer.