MICA: Development of a rapid in-vitro multi-drug test device to predict response to combined drug chemotherapy in leukaemia patients, before treatment

This project is to develop a rapid and inexpensive screening device that predicts how a patient will respond to leukaemia chemotherapy within hours. The test will initially be used for Acute Myeloid Leukaemia (AML), a potentially fatal cancer in which white blood cells multiply rapidly and unchecked in the bone marrow and blood. AML patients are usually treated with the anticancer agent cytarabine (Ara-C) in different doses or combined with other agents including Fludarabine, to increase the effectiveness of Ara-C. Up to 40% of patients do not respond to first course of chemotherapy treatment; lack of response is only seen after weeks when a patient may have suffered severe side effects. The nature of AML means it is vital to treat it as soon as possible; a delay of weeks can have severe consequences. This project will develop a quick and simple multi-drug test device based on blood or bone marrow samples BEFORE the start of treatment, to show response to COMBINED drugs used for treatment. The results of the multi-drug test will ensure that a patient receives the right combination of chemotherapy drugs, in the correct dose, to meet their needs and so prevent any delay in effective treatment. The test device will be particularly useful to patients whose leukaemic cells are sensitive to low doses of chemotherapy, allowing them to be treated with minimum drug doses, with less side effects. For elderly patients, where AML incidence and mortality rates are increasing, and where other health conditions may rule out aggressive chemotherapy, this screening test could be particularly timely. We have previously devised, published and patented details of a rapid test to measure response of AML patient blood or bone marrow samples, within 8 hours, to the SINGLE drug Ara-C. The test uses a bioluminescent bacterial biosensor that has been genetically modified to give increased light output in response to the active form of Ara-C within leukaemic cells; it gives an accurate measure of drug uptake and conversion to the active drug form. We have used our biosensor test on 71 patient samples and shown good agreement with a standard laboratory cytotoxicity test and a test efficiency of 83% when compared with 53 known patient outcomes. The single drug test will be extended to include other commonly used chemotherapeutics and the amount of patient cells per test reaction will be minimised, so that multiple drugs can be tested on a small clinical sample, vital for bone marrow testing, where sample volume is limited. Preliminary studies indicate that our device predicts the effectiveness of combined Ara-C/fludarabine chemotherapy. The test device will be extended to include other relevant drug combinations to exploit its full usefulness for AML treatment. The main aims of this project are to find the optimum conditions for a multi-drug test device that will predict how a patient responds to clinically relevant doses of combined drug chemotherapy, within 12 hours, using minimal number of patient cells. Also to validate the test device by retrospective testing of 500 clinical samples to correlate biosensor assay results with patient outcomes. This will produce a test device with a simple protocol suitable for routine laboratory use, which will be brought to 'regulatory approval' stage. The main application of our test device is in AML, to rapidly predict effectiveness of combined drug chemotherapy BEFORE treatment begins. Benefits include determining the most suitable treatment for an individual patient. Those with cytarabine resistant cells can be given effective combined chemotherapy that will enhance survival outcomes; patients with responsive cancer cells can be effectively treated with lower drug dose, reducing acute and long-term side effects including severe infection, infertility, secondary cancer, whilst enhancing quality of life and saving costs of hospitalisation.