Morning Ireland

Cancer Research in Ireland

There has been an explosion in the quantity and quality of cancer research in Ireland. Each of our universities now has high quality programmes in some aspect of the study of malignant disease including research into cancer biology, the diagnosis and monitoring of malignant disease and/or the discovery of new treatments. These are funded by the Irish Cancer Society, the Health Research Board, Science Foundation Ireland and philanthropy. Below is a flavour of some of the projects that are ongoing. For further information contact the Irish Cancer Society.

Prostate Cancer Consortium

Prostate cancer is a common cancer and a frequent cause of death in Irish males. Critical to improved diagnosis and better treatment for men with prostate cancer is the requirement for a more complete understanding of (i) what makes prostate cancer cells different from normal prostate cells (ii) why do some patients get very aggressive cancer while other men have a mild slow growing form of the disease (iii) can we exploit the abnormal biology of the disease to devise new treatment strategies. Understanding what genes and pathways may be damaged in prostate cancer cells can help us to translate this information to earlier diagnosis and better treatment options for men with prostate cancer.

The Prostate Cancer Research Consortium (PCRC) was established at the end of 2003, through a significant competitive grant from the Irish Cancer Society, with the stated aim of linking "like minded" researchers from different academic institutions and hospitals in a co-ordinated approach to maximize research potential in this disease. Bringing together researchers from different centres and different disciplines with complimentary "added value" skills provides an excellent environment to tackle important questions in relation to prostate cancer diagnosis, prognosis and new therapies. Our aspiration was to make a significant contribution to prostate cancer research, at all times with a clear focus on the cancer patient, while providing a teaching and mentoring environment that would encourage young scientists to flourish. Currently scientists, clinicians and allied healthcare professionals from Trinity College Dublin, University College Dublin, Royal College of Surgeons in Ireland, Dublin City University and their allied teaching hospitals (Mater Misercordiae University Hospital, St James's Hospital, Beaumont Hospital and the Adelaide and Meath Incorporating the National Childrens Hospital) are active members of the Consortium

Key events and discoveries within the PCRC include:
. Establishing the first Irish Prostate Cancer BioResource, containing samples taken from prostate cancer patients with their informed consent; today, samples from over 530 patients have been collected, providing a value resource for specific projects within the Programme of Research.
. Achieving novel and relevant research findings, which have been recognized by research awards at national and international conferences (see below) and published in the international literature (eg Cancer Research, Human Molecular Genetics, Br J Cancer, Endocrine Related Research).
o Novel genetic changes have been identified, which may increase your risk of getting prostate cancer.
o Particular biomarker combinations in prostate cancer cells have been detected, indicating a more aggressive form of the disease.
o Potential new targets have been found and new experimental approaches developed, which may allow new treatment approaches to be devised.
o PCRC researchers have been increasingly successful in attracting research funding from national and international agencies (eg Science Foundation Ireland, Health Research Board, Enterprise Ireland, Irish Cancer Society, Irish Research Council for Science, Engineering and Technology British Urological Foundation, European Union)
. Mentoring of a cohort of young scientists, clinicians and allied healthcare professionals as future leaders, who have demonstrated their ability through significant national and international awards including:
o Best Oral Presentation, Irish Association for Cancer Research 2006
o Best Oral Presentation, Irish Society of Urology, 2007, 2009
o Best Poster Presentation, International Cancer Conference 2008, 2009
o Best Poster Presentation, Association of Radiation Research, 2008, 2009
o Best Oral Presentation, National Cancer Institute/Dublin Region Higher Education Alliance Cancer Genomics MasterClass, 2008
o AstraZeneca International Scholar-in-training Award, 2009
o Best Poster Presentation, International Society of Cancer Gene Therapy, 2009
o St Lukes Young Investigator Award, 2009
. Publication of a comprehensive survey of prostate cancer patient's attitudes to biobanking, which showed a >85% very positive response and emphasizes the patient- centred approach of the Consortium

These promising advances have been achieved by the truly collaborative nature of the Consortium and the quality of the young researchers who have been nurtured through the programme, delivering science that has been recognized as high quality by international leaders and key journals in cancer research. The PCRC is setting an ambitious agenda for the next 5 years, to ensure that the quality of the research continues to improve, that the learning experience for young researchers makes a key contribution to their career development and that the patient continues to be at the centre of a collaborative, transdisciplinary research effort.

Cancer Research at TCD

Cancer is one of four strategic research priorities for TCD. It is an area where Trinity has significant quality expertise as judged by the publication of key papers in high quality international journals, the achievement of significant research funding from national and international agencies, the patenting of important discoveries and a clear path for the translation of key research discoveries to patient benefit, both at the level of improved diagnosis and prognosis and in the development of new therapies for cancer. TCD has a vibrant interdisciplinary research programme focusing on understanding the molecular and cellular biology of cancer but with a clear emphasis on the use of this information to benefit the cancer patient. Complimentary and added value strengths through researchers in the Schools of Medicine, Biochemistry and Immunology, Pharmacy and Pharmaceutuical Sciences, Genetics and Microbiology and Chemistry have allowed the development of a true interdisciplinary approach to cancer research at TCD. Translation of discoveries from the bench to the bedside is facilitated through close linkages to Institute of Molecular Medicine, the Cancer Clinical Trials Unit at St James's Hospital (the largest in the country) and the relevant clinical departments at St James's Hospital (St James's Hospital is the largest Cancer Hospital in the country). Translational research for patient benefit will be further strengthened by the opening of the Welcome Trust - HRB Clinical Research Centre in 2010/2011. A crucial bridge linking cancer discovery research and cancer clinical practice has been the establishment of cancer biorepositories, biobanks of relevant tissue and biofluids from the cancer patient carefully collected with informed consent and linked to clinical information. This resource allows new diagnostic and prognostic tests and novel potential drugs to be tested on relevant clinical material. Integrated programmes of research in cancer genetics, cancer cell biology, cancer immunology, cancer chemical biology, computational biology, experimental cancer medicine and cancer clinical trials have led to a range of key discoveries and developments including:

. The establishment of cancer biorespositories in key malignancies including prostate, lung, oesophageal, haematological malignancies, gynaecological malignancies, pancreas, colon etc
. The development of a novel gene based test for lung cancer
. The advancement of a novel cancer drug (Pa-Trin) which has gone into clinical trials
. A molecular test to evaluate the success of bone marrow transplantation for leukemia, now the test of choice in clinical laboratories throughout the world
. The fastest accruing cancer clinical trial in Europe at the time, for a drug that targets the abnormal biology of a particular form of cancer
. An understanding of the immunology of cancer, which is leading to the development of novel cancer vaccines
. A precise identification of the key events that cause cancer cells to survive, providing new targets and approaches for cancer therapy
. A clear understanding of the link between obesity and cancer
. An interdisciplinary approach that has identified a new type of drug which may have potential in the treatment of the most common form of leukemia

Cognisant of the need to develop the best possible structures to deliver high quality cancer research, TCD is currently establishing a Centre for Cancer Drug Discovery as part of a new Biosciences Institute which will open in 2011, providing an interdisciplinary approach to fast track important basic science discoveries to clinical translation. TCD is the first European university to be formally accepted to join the US National Cancer Institute's Chemical Biology Consortium, a significant development in relation to TCD's Cancer Drug Discovery Programme. A significant partnership has been established with the Centre for Cancer Research & Cell Biology in Queens University Belfast, underpinning the delivery of a critical mass of focused cancer research on the Island of Ireland.

Microneedle-based technologies that could help to treat cancer more effectively

Staff at the Tyndall National Institute and Cork Cancer Research Centre are developing new microneedle-based technologies that could help to treat cancer more effectively.

The treatment centres around electroporation, a technique whereby a brief electric pulse is applied to the tumour, thereby rendering cell membranes transiently porous, and making feasible the cellular uptake and efficacy of anti-cancer therapeutic agents. Electroporation is effective regardless of histological tumour type and cancers such as breast, prostate, kidney, melanoma, and colon can be treated.

Current methods of electroporation require the use of inefficient and sometimes invasive electrodes, but the of microneedle-based electrodes has the potential to simultaneously enhance treatment efficiency while reducing patient discomfort. Microneedles are sharp-tipped, micron-scale projections that painlessly puncture the skin but do not penetrate deep enough to stimulate the nerve endings; their application is therefore painless to the patient.

Teams led by Dr. Declan Soden (CCRC) and Dr. Conor O'Mahony (Tyndall) have already demonstrated promising results and further development of the technology is underway.

Molecular Therapeutics for Cancer Ireland (MTCI)

Lead Institution: Dublin City University
Partner Institutions: University College Dublin, Trinity College Dublin, Royal College of Surgeons in Ireland

The Molecular Therapeutics for Cancer Ireland (MTCI) (www.mtci.ie) research programme, which was launched in October 2009, focuses on aiding in the development of novel molecularly-targeted therapies for breast cancer and prostate cancer, while also providing a platform approach for research in other cancer types.

A key focus for Professor Gallagher's research within MTCI is the examination of the molecular basis of resistance to the commonly used hormone therapy, Tamoxifen, as well as to the leading antibody-based therapeutic, Herceptin. His group has specialist expertise in the use of in-depth functional interrogation approaches at both in vitro and in vivo levels, the latter benefiting in particular from the use of advanced molecular imaging technologies, such as positron emission tomography (PET) and optical imaging.

Professor Joe Duffy's research in this SRC aims to develop new treatments to prevent the spread or metastasis of breast cancer. The formation of metastasis is the main cause of death in patients with breast cancer as in other cancers. If metastasis could be prevented, death rates from cancer would be dramatically reduced.

Professor William Watson's focus is around the development of androgen independent prostate cancer and advanced disease. This project will use novel bioinformatics analysis of gene expression data generated as part of the work of the Prostate Cancer Research Consortium.

Cancer Research at University College Dublin

Developing the cancer biomarker pipeline
Researchers in UCD Conway Institute of Biomolecular & Biomedical Research are making novel discoveries in cancer biomarker development through the use of emerging technology platforms underpinning translational science and by forging exciting synergies across seemingly disparate areas of biology, mathematics and computing.

Introduction
Much of the current cancer research in UCD Conway Institute is focused on biomarkers of the disease itself or response to anticancer drug treatment regimes. These biomarkers may detect specific alterations in cancer cells, identify patients most likely to respond to drug therapy, help to monitor treatment efficacy or identify early signs of toxicity to drug treatment.

At present, many of the existing biomarkers of cancer can identify the disease at an advanced stage. Research is now focused on identifying and validating candidate biomarkers of early stage disease, prior to metastasis when intervention delivers improved prognosis for the patient.

Current thinking has also moved to identifying biomarker panels to overcome some of the shortcomings associated with individual biomarkers used in isolation such as lack of specificity and being able to detect very low levels of serum biomarkers.

Biomarker pipeline
UCD Conway Institute researchers work along the continuum of the biomarker discovery pipeline, which involves three main stages; the discovery of candidate biomarkers through any investigation of changes in gene or protein expression in cancer; analysis of candidate biomarkers in large sample sets to statistically verify their biological significance; validation of the biomarkers in prototype assays for use in the clinical laboratory setting.

The process of bringing a candidate biomarker along this pipeline from discovery to clinical use is challenging from many perspectives; technical, scientific, regulatory and funding. At UCD Conway Institute, researchers are addressing these challenges. The research of Conway Fellow, Professor R. William G. Watson in the area of prostate cancer is one example of how the multi-disciplinary approach to biomarker discovery at the Institute is working.

Prostate cancer biomarkers
Prostate cancer is the most common malignancy among men in developed countries and is associated with significant morbidity and mortality. In the Republic of Ireland, the current annual incidence is approaching 1,700 and mortality figures are in excess of 550 cases per annum.

Professor William Watson is part of the Prostate Cancer Research Consortium (PCRC), an initiative established under Molecular Medicine Ireland to improve the detection, prognosis and treatment of the disease. A key feature of this consortium is a prostate cancer bioresource; a collection of clinically annotated biospecimens, which provides the vital material for research projects.

No single biomarker exists that can accurately diagnose the presence of prostate cancer, identify the extent of the disease and predict how it will progress in an individual patient.
Current screening methods rely on testing serum for elevated levels of prostate specific antigen (PSA). However, PSA is not specific to prostate cancer and can be seen in benign conditions such as prostatitis.

Contemporary prostate cancer research is turning to identifying panels of protein biomarkers that can act as prognostic tools in parallel to clinical evaluation. This could allow appropriate surgical intervention and improve patient outcomes.

Professor Watson and his group identified a novel panel of serum markers for the progression of prostate cancer using a powerful protein separation technique, 2-dimensional-difference gel electrophoresis (2D-DIGE) in the Proteome Research Centre of the Institute. This research was published last year in the Journal of Proteome Research1.

The work of analysing the vast quantities of data produced from the discovery phase of this research is being carried out by a doctoral student of bioinformatics and computational medicine being jointly supervised by William Watson- an example of the opportunities available to Conway researchers to capitalise on synergies in seemingly disparate areas of the sciences.

This research has now advanced in the pipeline to the validation stage. The American Association Cancer Research (AACR) recently recognised the research to validate this novel panel of protein biomarkers at a special conference focused on prostate cancer in San Diego, California. This biomarker panel will now go into a larger, international validation study in collaboration with consortium partners in Austria, Australia and the USA.

Breast cancer biomarkers
Professor Joe Duffy and his colleagues at St Vincent's University Hospital, Dublin have already validated a breast cancer biomarker, urokinase plasminogen activator (uPA) and its inhibitor, PAI-1, as prognostic markers in breast cancer through large international studies2. One of these studies involved in excess of 8,000 patients from 18 different European hospitals, including St Vincent's University Hospital.

Several expert groups including The American Society of Clinical Oncology, The National Academy of Clinical Biochemistry (USA) and the European Group on Tumour Markers stated that the uPA test may be used for determining prognosis in newly diagnosed breast cancer patients, especially in those women who present with lymph node-negative disease.

The uPA biomarker is from a family of proteases that digest cell barriers and allows tumour cells to metastasise. High levels of uPA point to a stronger likelihood of the patient developing metastasis and requiring chemotherapy as well as surgery to fight the disease. However, if the levels of uPA are found to be low, a combination of surgery and radiotherapy may be sufficient in order to treat the cancer, helping to minimise the expense and the unpleasant side effects of the treatment, for the patient.

Professor Duffy is now collaborating with UCD Conway colleagues, Professors William Gallagher and Finian Martin to develop a panel of biomarkers that will further enhance predictive accuracy and improve treatment regimes and clinical outcomes for patients.

Technological advances creating opportunities

Breast cancer
The numbers of candidate biomarkers emerging from large-scale screening approaches at the discovery stage of the biomarker pipeline has created a major bottleneck in the cancer biomarker development process at the validation stage.

Professor William Gallagher and his research group are leading the way in addressing this bottleneck through the application of tissue microarray (TMA) and digital imaging technologies to oncology, with a focus on breast cancer biomarker development.

Tissue microarrays (TMA) are cores of breast cancer tissue mounted in paraffin blocks and processed onto glass slides. This technology provides a high-throughput platform for pathological investigation, allowing analysis of several hundreds of specimens simultaneously on a single slide. In combination with digital slide technology, the interpretation of histopathological and immunohistochemical specimens can be made using automated image analysis algorithms. The added advantage of this digital technology is the ability to share information at distant sites.

In a recently published paper in Breast Cancer Research3, this group automatically quantified expression of two standard clinical markers used in breast cancer assessment, oestrogen (ER) and progesterone (PR) receptors in over 700 patient samples from 2 different cohorts using this image analysis approach.

The finding that automated quantitation of ER and PR immunohistochemical data compared favourably with manual scoring results has key implications for fast-tracking the validation of biomarkers. Enterprise Ireland has recently awarded a technology development grant to William Gallagher in order to deliver a clinically useful decision support tool for pathologists.

Bladder Cancer
Using similar TMA technology, Dr Amanda McCann recently identified a novel biomarker and potential therapeutic target for urothelial carcinoma of the bladder (UCB), in collaboration with clinician colleagues, Professor John Fitzpatrick, Mater Misericordiae University Hospital and Professor Elaine Kay, Beaumont Hospital and Royal College of Surgeons in Ireland.

The majority of bladder malignancies do not invade muscle at diagnosis. However, these tumours have a highly unpredictable potential for recurrence and progression into muscle invasive disease. High-grade UCB tumours with lamina propria invasion (pT1, grade 3) represent those at the greatest risk. There is much debate about the appropriate surgical management of these patients. Nearly one-third will ultimately require cystectomy as second-line treatment. By identifying these high-risk cases early, the appropriate radical treatment can be given from the outset.

In a recent publication, the group have shown that immunohistochemical assessment of CDH1 staining can independently predict time to recurrence in the non-muscle invasive pTa and pT1 tumours over and above stage and grade. In addition, CDH1 cytoplasmic staining is predictive of recurrence in the controversial pT1, grade 3 group.

IGF-II loss of imprinting (LOI) has already been identified as an epimarker of colorectal cancer development. This research suggests that the finding of LOI in the tumour associated normal of this UCB cohort points to it as a predictor of tumour development and playing a significant role in compromising cellular adherence through the internalisation of CDH1.

Dr Amanda McCann believes that including specific assessment of cytoplasmic CDH1 staining might be beneficial in the clinical setting in the future to determine whether the bladder should be conserved in high risk patients. She believes the possibility of impeding tumour progression by restoring aberrant clinical levels of IGF-II to normal is an exciting one.

Glycobiology opens new horizons
The arrival of Conway Fellow, Professor Pauline Rudd and the establishment of the Dublin-Oxford glycobiology laboratory in UCD Conway Institute has added another facet to the research on cancer biomarkers.

One of the most common post-translational modifications to proteins that occur is the addition of sugar(s) during glycosylation. There are a huge range of possible sugar conformations that result. Glycan processing pathways become disturbed in cancer cells so monitoring the glycan signature of proteins may potentially provide a powerful way of diagnosing and tracking the course of the disease.

The high-throughput technology platform for glycan alanysis developed by Professor Rudd and her team is based on high performance liquid chromatography. This sensitive and quantitative technique has made it possible to simultaneously screen a single blood sample (<5 microlitres of serum) for many glycoproteins.

Rudd and her team believe that the greatest potential source of easily accessible biomarkers are the cell surface proteins shed from tumours and individual proteins secreted from cancer cells into body fluids such as serum. Their collaborative research with other Conway scientists and clinicians is highlighting how glyco-biomarkers of cancer could play an important role in the development of the biomarker pipeline from discovery to validation.

Some of the current projects in their cancer programme include looking at acute phase proteins with specific glycan changes correlating with advanced ovarian cancer5; signalling pathways that underpin glycosylation changes in breast cancer cells; identifying low level glycoprotein signatures of cancers including prostate and ovarian as well as glycan changes indicative of staging gastric and pancreatic cancers.

Their technology programme is not solely focused on improving high throughput screening at the discovery phase. They are currently developing an ELISA-based assay to detect changes in glycosylation associated with free PSA in prostate cancer that would be suitable for commercial development and the clinical setting. Pauline Rudd is also a principal investigator in the National Institute for Bioprocessing Research Training (NIBRT). Her high-throughput technology has valuable industrial applications where pharmaceutical companies need to monitor glycosylation patterns in the course of drug production.

Future directions in the biomarker pipeline
Professor Dolores Cahill came to UCD Conway Institute in 2005 after eight years at the renowned Max Planck Institute, Germany where she accumulated the largest collection of proteins (10,000) on array. In collaboration with Professor John O'Leary from the Coombe Women's Hospital and Trinity College Dublin, Professor Cahill plans to examine the possibility of blood based assessment of early & late stage ovarian carcinoma.

Cancer cells secrete proteins that leak from the tissue into the bloodstream and cause an autoimmune response similar to that seen in vaccination. The secreted proteins exist at low levels in the blood stream during early stage cancer and so detection is an issue. However, the autoimmune response to the protein amplifies the signal and makes it easier to identify these antibodies in the serum of patients.

Dolores Cahill plans to use gene array technology for the discovery part of this planned biomarker pipeline. Samples of ovarian cancer can be analysed against her protein collection to uncover candidate biomarkers.

Cahill believes that this method for high throughput discovery has distinct advantages over others with the ability to detect cancer proteins at low levels to identify early stage disease and the potential to exploit an ELISA method of antigen-antibody detection for use in the clinical setting. However, the challenges she and her colleagues have faced in order to establish this research has highlighted the need for support mechanisms that allow adaptable responses to the possibilities afforded by evolving technologies.

Dolores Cahill highlights the need both for greater awareness of the value of bioresources with specific consent relating to use of samples for validating diagnostic biomarkers and sources of sustainable funding to facilitate the research process throughout the biomarker pipeline. Providing appropriate supports along the length of this biomarker pipeline may ultimately shorten the transit from the research laboratory to the clinical setting and improve the quality of the product to the benefit of patients.

Images

Example of automatic assessment of nuclear protein expression in breast cancer tissues A novel unsupervised clustering approach was used to overcome one of the major barriers in image analysis - the identification of tumour nuclei from stromal tissue. The output shows positive tumour nuclei in red and negative tumour nuclei in blue. Data generated from the analysis include % positive nuclei and nuclear intensity.