Lay Summaries

Dr Chris Womack – AstraZeneca

Application for tissue to support a programme of exploratory research projects that in turn, support and optimise the early stages of cancer drug discovery and development and also to gain information of relevance for design of clinical studies, e.g. "Proof of Mechanism, Proof of Principle and Proof of Concept". Research studies will lead to a better understanding of the natural history of cancer and may generate hypotheses on the predictive value of specific molecular features for the natural history of specific cancer types. This will offer the potential of identifying patients with those tumour types considered most likely to benefit from “molecularly targeted therapies”. Some of the studies will be carried out at AstraZeneca and some in collaboration with scientists in Wales. The application is therefore not for a specific project with defined research outcomes, but for acquisition of tissues to enable assessment of utility and reliability of marker detection by a variety of methods in a variety of cancers and normal tissues. The technologies used will have been tried and tested using cell lines where appropriate or will use antibodies and probe sets defined by others and reported in the literature.

The primary objective of the initial studies outlined here is to investigate molecular targets from selected cell signalling pathways that drive tumour cell proliferation, survival, angiogenesis and invasion. A secondary objective will be to correlated selected molecular biomarkers with clinico-pathological parameters.

Published paper
Smith, N; Baker, D; James, N; Ratcliffe, K; Jenkins, M;Ashton, S; Sproat, G; Swann, R; Gray, N; Ryan, A; Jürgensmeier, J; and Womack, C (2010), 'Vascular Endothelial Growth Factor Receptors VEGFR-2 and VEGFR-3 Are Localized Primarily to the Vasculature in Human Primary Solid Cancers', Clinical Cancer Research; 16(14) July 15, 3548-61

Dr Nessa Carey – CellCentric Ltd
Epigenetics, the reversible heritable changes in gene function that occur without a change in DNA sequence, is an area of outstanding interest in oncology. It is now widely accepted that cancerous cells display a range of epigenetic abnormalities and that these alterations represent changes which have functional impacts on cellular behaviour. There have been successes in the targeting of drugs to epigenetic pathways in cancer. Decitabine is a DNA methylation inhibitor licensed by the FDA in the treatment of myelodysplastic syndromes, and the histone deacetylase inhibitor, vorinostat, has been licensed for use in patients with cutaneous T-cell lymphoma.

Although the results with the drugs mentioned above are very encouraging, and provide proof of principle in the application of epigenetic therapies, they are unlikely to be the most effective epigenetic modifiers that will be developed. To generate more effective drugs in this field, we require a much greater understanding of the complex networks of gene regulation and how these are disrupted in disease. CellCentric are working with a variety of scientists who are approaching epigenetics from a mechanistic rationale, identifying key regulatory genes and proteins in defined cellular epigenetic pathways. The information gained from these studies is extended to analyse the impact of these genes in abnormal epigenetic events in cancer, such as aberrant silencing to tumour suppressors. A number of candidate oncology epigenetic genes have been identified by this approach and we now wish to move into more advanced target validation and therapeutic product development.

Dr John Staffurth - Cardiff University
To characterise local and systemic immunity in prostate cancer and the immunological implications of androgen deprivation therapy (ADT), leading to the rational design of novel immunotherapy of prostate cancer in conjunction with existing therapies.

Dr Rebecca Porter - Cardiff University
To test whether differences in activation of the EGF, Notch and Wnt pathways could explain the differences in growth patterns observed in basal cell carcinomas in vivo. This information will be critical to the delivery of effective topical therapy for basal cell carcinoma.

Professor Horst Zitzelsburger - GSF-National Research Centre for Environment and Health, Germany
To identify genetic aberration patterns in human breast cancers with RET expression or RET/PTC rearrangement in comparison to tumours without RET expression. This will uncover molecular mechanisms related to RET expression during breast cancer development. Further studies could then relate the observed aberration patterns with clinical outcome and identify genetic markers which are of prognostic relevance in these tumours.

Dr Michaela Aubele - GSF-National Research Centre for Environment and Health, Germany
In this study we aim to perform protein profiling using 2D gel (DIGE) and mass spectra on primary breast carcinomas. Analysis will be performed comparing the protein expression pattern of triple-negative tumours (Her2- ER- PrR-) versus tumours being positive for one of the parameters, respecitvely (Her2- ER+ PrR-/Her2- ER- PrR+/Her2+ ER- PrR-). The identified differentially expressed proteins will be characterised and analysed for their potential as a new therapy targets using data bases.

Published paper
Schulz, D; Bllner, C; Thomas, G; Atkinson, M; Esposito, I; Höfler, H; Aubele, M (2009), 'Identification of differentially expressed proteins in triple-negative breast carcinomas using DIGE and mass spectrometry', Journal of Proteome Research 8 (7), 3430–3438

Dr Simak Ali – Imperial College, London
Prostate-Specific Antigen (PSA) testing is widely used in the diagnosis of Prostate Cancer and in the monitoring of treatment for the disease. However, although the test is comparatively simple, PSA testing fails to identify a significant proportion of aggressive cancers, while only about a third of men with a 'positive' PSA test have tumour. Additionally, of men who are initially treated for prostate cancer, about a quarter require additional treatment, presumably due to recurrence of the disease. Together, these findings highlight the urgent need for new tests, which not only detect prostate cancer but can also help distinguish indolent from aggressive disease. In order to identify new, more specific diagnostic markers, we have been studying the changes that androgens, the hormones that drive prostate cancer growth, bring about to the types and amounts of proteins made by prostate cancer cells. One particular protein was made to very high levels upon treatment of prostate cancer cells with androgen. This protein is found in body fluids, including serum at low levels and is easy to detect. We now propose to evaluate this protein as the basis of a new diagnostic test for prostate cancer, by measuring levels in patient sera. We will also carry out experiments on prostate cancer cells grown in the laboratory to see if the protein might play a role in development of the disease itself. Collectively, the potential benefits of this research for men affected by prostate cancer will be an advance in our understanding of the mechanisms underlying prostate cancer development and the development of a new test for prostate cancer diagnosis.

Dr Duncan Baird – Cardiff University
Telomeres prevent the natural ends of chromosomes from fusing; chromosomal fusion can lead to genomic instability that may drive the earliest-stages of cancer. It is considered that many different types of cancer can be initiated by telomeric instability and fusion. We have developed technology, which allows us to detect telomere fusion events from single DNA molecules. Our data from cells grown in vitro provides a definition of the length at which telomeres lose their protective function, and provides insights into the mechanism that may cause fusion. Importantly, we have also shown that in normal human cells, telomeres can suffer sudden large-scale deletion events, which created severely shortened telomeres that can then fuse. Thus telomeric deletion and fusion may represent the earliest genetic lesions that can lead to cancer; we therefore wish to establish if our telomeric fusion assay could be useful as prognostic/diagnostic marker. Before we can consider doing this, we need to ascertain if telomere fusion is present in tumour samples. If we can detect telomere fusion in these samples it will inform us about the biology of this mutational event, but importantly will provide evidence that will allow us to start to look for these events in early-stage lesions.

Dr David Millar, Cardiff University
To date, the majority of breast cancer researchers use immortalized cell lines. These cell lines are prone to genetic drift and, in the majority of cases, have been derived from metastatic deposits and not the primary tumour. Thus, a more clinically relevant model would be the use of epithelial cells derived directly from primary breast tumours, as the majority of treatments used today are targeted against the primary tumour. The primary culture of tumour derived cells will therefore be useful for both functional cell and molecular biology studies. To this end a culture system will be established based on the methods of Speirs (2003).

Professor Gerry Thomas, Imperial College, London
The ret oncogene is known to be associated with specific types of thyroid cancer. Recently is has also been found to be expressed in breast cancer. It is activated by two different mechanisms in breast cancer – by increased expression and by rearrangement of the gene, which results in fusion of part of the end of the gene and an increase in downstream signalling pathways. This project aims to relate expression/rearrangement of the ret gene to specific molecular phenotypes of breast cancer, and to identify the fusion partners when the ret gene is activated by rearrangement in breast cancer. Many drugs that are currently used in cancer treatment to block the development of new blood vessels in tumours also inhibit the signalling pathway for the ret oncogene. The results of this study may lead to identification of subsets of breast cancer patients who may benefit more from these types of treatment regimes than others who do not show activation of the ret gene in their tumours.

Dr Christian Rohlff, Oxford Genome Sciences (UK) Ltd
Oxford Genome Sciences (OGeS) will use samples from the Wales Cancer Bank to establish tests for the diagnosis of cancer and for monitoring patient relapse and progression. OGeS has recently discovered a panel of membrane-associated, transmembrane and secreted proteins that are found to be up-regulated in cancer patients, and to have low normal expression. These proteins were discovered via proteomic detection and prioritised using the company’s OGAP® data integration system. OGeS has evidence that some of these biomarkers are likely to be up-regulated in the blood of cancer patients and may therefore provide specific and sensitive diagnostic biomarkers for the early detection of cancer and for the monitoring of disease progression and relapse. OGeS intends to conduct an investigation with blood serum samples from the Wales Cancer Bank to confirm whether these proteins are indeed up-regulated in the blood of cancer patients. If successful, the biomarkers will be taken forward for further investigations for the development of rapid and sensitive tests for prostate, breast and colorectal cancers.

Professor Alan Burnett, Cardiff University
Prostate cancer is thought to originate from cells that are capable of self-renewal and the ability to specialise into many different cell types; these cells are known as cancer stem cells. Although conventional chemotherapy is able to reduce the size of prostate tumours it is thought that the continued presence of cancer stem cells results in re-growth and spreading of the tumour. Prostate cancer cells have high levels of a protein called NF-kB which is important for the survival of the cancer cells. Recently we demonstrated that an inhibitor of NF-kB is effective at killing leukaemia cells and myeloma cells, and others have shown that it preferentially targets the leukaemia stem cells. We now intend to use this inhibitor in prostate cancer samples to determine whether it is also an effective treatment in this disease and whether it is effectiveness is maintained in prostate cancer stem cells. In this way we hope to develop better treatments for this disease.

Dr Lisa Spary, Cardiff University
We are interested in the effects of androgen deprivation therapy (ADT) on the immune system of patients diagnosed with prostate cancer (PCa). In order to analyse the effects of ADT we require PCa cells for use in the laboratory to establish PCa cell lines for further research. It is extremely difficult to grow prostate cancer cells in the laboratory and most PCa cell lines available are derived from PCa that has spread throughout the body. We therefore wish to grow PCa cells, from the original PCa in the aim to develop PCa cell lines that can be used in our experiments. Once numerous cell lines are established we will perform multiple experiments to characterise the cells and also see the effect of hormones on these cells. We are currently establishing a method which enables us to grow sufficient numbers of primary PCa cells from small biopsies for experiments in the future.

Drs Dimity Pshezhetskiy, Justin Stebbing and Leandro Castellano, Imperial College, London
We propose to investigate a novel mechanism used by prostate cancer to spread to other parts of the body like the bone. This process is called metastasis and is the major cause of death from prostate cancer. Metastasis is currently an incurable aspect of the disease. In our recent work we have discovered a new molecular event in prostate cancer cells that could allow them to spread by the process of metastasis. These are particularly exciting results because drugs that can block this molecular event are available so could easily be tested in patients in clinical trials to see if they can stop their metastasis.

Dr Claire Aukim-Hastie, Portsmouth University
Prostate cancer (PCa) has an extremely variable outcome with apparent indolent (slow growing) and aggressive forms of the disease and survival periods ranging from 2 to >15 years. Therefore there is a pressing need to enhance the early detection and appropriate management of prostate tumours. At present however, prognostic information for PCa is extremely limited and the markers in use are unreliable. Using protein analysis methods to study PCa patient serum samples, we have carried out a pilot study to identify proteins associated with the clinical stage of disease. This pilot study identified 26 putative markers of progression with 3 of those markers capable of predicting aggression of the disease based on Gleason score. Additionally, 16 potential biomarkers have also been found that correlate with recurrent disease with one of those capable of predicting recurrence based on prostate specific antigen (PSA) level and metastasis. We aim to validate these findings by studying a geographically distinct population of PCa patients using the same technique. In addition, we aim to further validate these potential biomarkers using both sample sets but utilizing different protein analysis methods.

Dr Jane Wakeman, University of Wales, Bangor
Colorectal cancer is a major cause of death throughout the western world. A particular feature in some tumours is the ability of a sub-group of cells, tumour initiating cells, to leave the primary tumour site and establish a new tumour at a more distant site within the body, a process known as metastasis. Such movement of cells from the primary site and re-establishment at a secondary site occurs in the more aggressive tumours and involves the dynamic regulation of factors that are not normally expressed in the tumour. Using a laboratory model for this dynamic process, we have identified a factor, which may be important in maintaining such tumour initiating cells in colorectal tumours. We would now like to extend our data from the laboratory model to determine the expression patterns of this factor in patient derived, colorectal tumour samples. This will allow us to assign significance to our lab based data, in terms of the importance of this factor in generating new tumours.