John van Geest Cancer Research Centre
Tumour Immunology and Molecular Genetics
The research conducted by the Tumour immunology and Molecular Genetics group focuses on the identification of new tumour-associated proteins, their expression pattern and function as well as the identification of MHC class I/II- peptides derived from those antigens and the assessment of new vaccine strategies.
The team of 13 research staff collaborate with major European Research Institutes engaged in research to develop and apply cancer vaccines. The research is multidisciplinary, involving cellular / serological assays to detect tumour immune responses, molecular biology and biochemical methodologies as well as clinical proteomics using analytical mass spectrometry in conjunction with Artificial Neural Network (ANN) data mining.
Antigen Discovery:
We have applied the SEREX approach to identify novel tumour antigens (T21, T128 and Pr genes). We have also developed array technologies which allow us to rapidly screen multiple expressed gene products from cDNA libraries against individual sera from patients with cancer, benign prostatic hyperplasia (BPH) and normal control subjects.
Using Expressed Sequence Tag (EST) database mining, we have identified four novel breast-associated UniGene clusters named BUC.
Molecular and Functional Characterisation:
We have used semi-quantitative RT-PCR and RT-qPCR on laser capture microdissected materials to evaulate mRNA expression patterns of all our recently discovered genes and some selected known genes using a panel of RNA from normal and malignant tissues.
We have also initiated a programme of research to examine the tumour cells' requirement for their expression. We have specifically silenced each gene and assessed the effect this has on tumour cell survival/proliferation.
We are also working on HAGE (a member of the DEAD-box family of ATP-dependent RNA helicases whose function so far remains unknown. HAGE is a cancer-testis antigen and we have already shown it to be immunogenic and to be an effective target for the immune system in vivo.
By clarifying the function and regulation of these genes in tumour cells or at least the signalling pathways where they are involved we aim to be able to specifically target them using novel or existing treatments.
Peptides identification / vaccine design:
We have identified several immunogenic and endogeneously processed peptides derived from p53, PAP, HAGE, Tyrosinase and BCR-ABL proteins. Peptides are poorly immunogenic and must therefore be co-administered with the immune potentiators (adjuvants), which will trigger early innate immune responses, which in turn are crucial to achieve vaccine effectiveness.
Our current collaborators include The Urology Centre, NUH, Nottingham, The Radium Institute, Oslo, Norway and the Hospital George Pompidou, Paris, France.
In collaboration with colleagues in Paris and Nottingham, we aim to compare the efficiency of Shigella toxin and Immunobodies to deliver and stimulate the immune system of prostate cancer patients in a Phase 0 clinical trial. For this, immune monitoring will be extremely important and techniques such as ELISPOT and multi-colour parameter flow cytometry are invaluable.
