The ECMC is currently supporting the NCRI AML16 and 17 trials in AML. These are the largest trials in AML in the world and are lead from Cardiff with about 150 centres in the UK Denmark, New Zealand, Australia and Norway. Diagnostic material for all of AML16 and 50% of AML17 is banked in Cardiff running at about 600 samples per year. Cardiff undertake mutation screening in the both trials for FLT3 mutations (n=650 in 2009). This enabled patients to be randomised to inhibitor or placebo. This is the first first-line intervention randomised trial. Interim data oral presentation at ASH meeting December 20091 (Knapper et al BLOOD 2009 abstract).

With archived material we can investigate potential biomarkers. For example we refuted the claims of a small study that mutations of NPM1 and MN1 over-expression predicted response to ATRA2 (Burnett et al BLOOD 2009 in press). We have produced some of the first data to show that NPM1 predicts a favourable outcome in older patients with AML, and this will be prospectively used in the AML16 trial to direct induction treatment. Samples are routinely screened for cKIT and NPM1 (n=650 in 2009). The Centre also undertakes serial monitoring of plasma inhibitory activity of the FLT3 inhibition in AML17, and of methylation status of patients on demethylation maintenance on the AML16 trial (n=55 in 2009).
We recently developed and incorporated a mathematical risk score for selection of AML patients for transplantation which operates in the AML17 Trial (Burnett et al BLOOD 2007), and have extended that to a score which predicts a 10% survival benefit if they are given the immunoconjugate Gemtuzumab Ozogamicin (Mylotarg), which includes 70% of younger patients. Both are being prospectively validated in the AML17 Trials and potentially in the US SWOG Trial.

Using the extensive Cell Bank which we now hold (>1200 characterised samples), we regularly test novel agents for activity against primary cells to identify potential agents to incorporate into early stage AML trials (see below). We have around 20 agents under assessment at any one time. These include modified nucleoside analogues from the McGuigan lab in Cardiff. We have agents from Novartis, Sunesis, Cyclacel, Leuchemix, Ambit, Antisoma, Chroma, under investigation with a view to making a case for AML clinical studies. These may well feed into out early stage trial strategy.

In house we have identified plausible druggable targets in AML derived from our GEP data and transfection correlates e.g. PDK1 and CD200. We have tested agents against PDK1 which could potentially be trial candidates. We have identified a humanised CD200 antibody and are embarking on discussions with the relevant company.

Part of the Cardiff ECMC was to establish a reliable and broad initial screen to filter out in-actives, and allow effort to be focussed on promising ‘hits’ primarily for solid tumours. A testing cascade is this in place starting with 4 established tumour cell lines, chosen to capture 9/10 of initial hits. Following this, full dose response curves are obtained, and secondary assays in resistant cells, or primary material, or with pharmacological probes included. In this way, the ECMC is feeding into a pipeline for future drug development. Links with Pharma are strong, particularly from the McGuigan and Burnett labs, and these are being exploited to enable onward funding and exploitation of early hits. The aim of this project is to establish a high throughput cancer cell culture pharmacological screening programme, capable of screening potentially up to 200 novel cytotoxic and anti-cancer compounds annually. To date a total of 271 compounds have been submitted, assayed across the cell line panel. Of these, 110 have been assayed in 2009. Based on this data a number of ‘second generation’ compounds have been submitted for ongoing testing.

The ECMC has linked to the Tenovus Cancer Research Centre in Cardiff University and the Breast Cancer Unit at Velindre Hospital via funding for a Grade 5 research technician based within the Tenovus Centre. The ECMC-funded aim is to build a clinical programme investigating new tumour markers and therapeutic agents in breast cancer stemming from basic laboratory studies undertaken in the Tenovus Centre. The Tenovus Centre has developed multiple human breast cancer cell lines which reflect this disease’s diverse genetic background and its progression towards an endocrine resistance state, where the cells are used to identify novel pathways that limit response or are associated with de novo and acquired endocrine resistance. The overall current schema employed by the Breast Cancer Team has 5 phases:

1. Candidate and non-candidate pathway identification in the breast cancer cell lines using modern molecular and cell biology approaches;
2. Pathway verification using molecular and pharmacological knockdown of identified pathway components in the models;
3. Development of robust immunohistochemical (IHC) assays for pathway markers in formalin-fixed paraffin embedded (FFPE) breast cancer specimens and their screening in samples from appropriate clinical trials and studies to establish clinical relevance in relation to endocrine treatment outcome and disease clinicopathology;
4. Interaction with Industrial Partners to explore the value of pathway inhibition in our panel of responsive and resistant cell lines using targeted pharmacological agents nearing or in clinical development;
5. Involvement with clinical trial development to assess the relevance of new tumour markers and to test new therapeutic approaches.

The ECMC funded technician currently is involved in the research areas outlined in phases (3) and (4) above, with the work involved in (4) being restricted to the testing of drugs with MTAs but no industrial funding.

Cardiff are the Hub on AML trials which run in >150 centres. The 2 national trials devised in Cardiff have a total of 15 randomised interventions several of which are completely novel and based on either developing a pre-clinical rationale or on previous phase ½ development lead from Cardiff. We anticipate the introduction of AC220 and possibly sepacytiabine and voreloxin to these trails via the “pick a winner” design that we developed for leukaemia. The pick a winner runs in about 80 centres and recruits about 250 per annum.