New in vitro culture conditions for primary malignancies that mimic in vivo conditions have been established and this has led to a highly productive year with identification and validation of new therapeutic targets through molecular cloning of novel chromosomal translocations in B-cell precursor acute lymphoblastic leukaemia and through proteomic analysis of the cell surface of mature B-cell malignancies. Three potentially important targets are CRLF2, HVCN1 and BCL6. CRLF2, the receptor for the cytokine TSLP, has been shown to be deregulated in a subset of BCP-ALL associated with poor prognosis. Therapeutic antibodies are currently being generated. HVCN1, a voltage gated proton channel expressed in B-cells, has been shown to regulate signalling via the B-cell receptor and is being considered as a therapeutic target for B-cell malignancies. BCL6 is a transcription factor involved in chromosomal translocations in diffuse large B-cell lymphoma and expressed in Burkitt’s lymphoma and follicular lymphoma. We are developing routes to producing small molecule antagonists of BCL6 and have already fully characterised one peptide aptamer, Apt48, that binds to the BCL6 POZ domain at residues D33-R44. Using in silico docking studies we have discovered compounds in commercial and other chemical libraries that potentially bind to this site. A major achievement of the group in 2009 has been the undertaking of clinical trials with a BCL2 inhibitor developed largely through the group’s laboratories. Phase I studies with the rationally-designed molecule ABT-263 in patients with chronic lymphocytic leukaemia (CLL) and in small cell lung cancer commenced in Leicester.

Research to discover and develop novel phytochemicals as potential cancer chemopreventive agents and/or as potential adjuncts to chemotherapy continued. Work was completed that will lead to a clinical trial of curcumin with oxaliplatin chemotherapy in advanced colorectal cancer. In vitro and in vivo studies have demonstrated an increase in efficacy of up to 300-fold for oxaliplatin by the addition of curcumin. Mechanisms for this activity are being identified. A single agent phase I study of curcumin opened in 2009, which has informed the appropriate formulation and dosing schedule for the combination study. A source of MHRA-acceptable drug has now been obtained and the protocol written and submitted for regulatory approval to open in early 2010. Other studies with phytochemicals carried out in 2009 include a phase I trial of a novel formulation of resveratrol (from Sirtris Pharma), with improved oral bioavailability as compared to unformulated resveratrol and this was shown to be safe and tolerable. PK and PD data are currently being analysed. Three studies with [14C]-labelled resveratrol were initiated in 2009 and involve healthy volunteers and patients with colorectal cancer or prostate disease. Accelerator Mass Spectrometry is being used to assess detailed PK of low, dietary-relevant doses compared with high pharmacological doses to provide new insights into the optimal dosing of potential chemopreventive phytochemicals.

Work analysing cell free plasma DNA of breast cancer patients on follow-up for evidence of disease progression has been completed. We have developed real-time quantitative PCR assays for measurement of circulating DNA quantity and integrity and to detect tumour specific changes such as HER2 amplification. Through collaboration with Imperial ECMC (Prof RC Coombes) we have shown statistically significant differences in mean plasma DNA concentration between cancer patients and individuals with both benign breast disease and healthy female controls (P <0.001). Results of work predominantly undertaken in 2009 offers the exciting possibility of developing a blood test to diagnose breast cancer and differentiate benign and malignant breast masses.

Examination of the relevance of Notch signalling in pancreas cancer with the aims of defining new biomarkers for diagnosis/prognosis and to develop novel therapeutic strategies. Nuclear localisation of Notch3 was found to be linked to a more aggressive tumour phenotype with the Notch pathway interacting with STAT and PI3K/Akt survival pathways. Subsequently we have found that Notch1, Notch4 and a downstream target, Hes1, are also overexpressed in the majority of resected pancreatic adenocarcinoma specimens when compared to normal pancreas. Treatment with a gamma-secretase inhibitor GSI-I results in a dose-dependent inhibition of Notch activation and a reduction in Hes1 expression, which is associated with induction of apoptosis and G2/M cell cycle arrest. Curcumin treatment, which inactivates Notch3 and 4, also results in a significant reduction in cell viability and induction of apoptosis. Combining GSI-I with gemcitabine or curcumin results in improved efficacy compared to single agent treatments. We commissioned an antibody to the Notch1 fragment and coupled it to magnetic beads to immunoprecipitate the peptide from culture medium. We subsequently detected (at femtomolar levels) a Notch1-related peptide in the serum of 7 out of 10 patients with advanced (non-resectable) pancreatic cancer. No such signal was obtained in 10 control samples from healthy volunteers. This suggests Notch may be a novel tumour marker in diagnosis of pancreatic cancer, which could aid decisions on treatment options and assessment of patient response.

Preclinical research involving the hepatobilliary team of the Leicester ECMC using eicosapentaenoic acid has demonstrated significant biological effects on angiogenesis, which led to investigation of its anti-tumour activity in pancreatic adenocarcinoma cell lines. Significant increases in cytotoxicity of gemcitabine were observed and a phase I/II clinical trial was initiated in 2009 combining eicosapentaenoic acid with gemcitabine in patients with newly diagnosed pancreatic cancer. To date the combination has been well tolerated and rapid recruitment should lead to conclusion of the study in early 2010 with the potential to undertake a randomised phase III study if results are promising. This offers the hope of improved outcome for patients with this disease and also the potential for improved quality of life as eicosapentaenoic acid has previously been shown to reduce weight loss and fatigue in patients with advanced disease.

Extension and refinement of a quality assurance system for GCLP in the laboratories of the ECMC groups with full roll-out occurred in 2009. This has enabled all clinical trials material to be analysed and used for registration and publication in the future.