A bioluminescent microbial biosensor for in-vitro pre-treatment assessment of cytarabine efficacy in leukemia
Salisbury, V., Anderson, E. and Alloush, H. (2010) A bioluminescent microbial biosensor for in-vitro pre-treatment assessment of cytarabine efficacy in leukemia. Clinical Chemistry, 56 (12). pp. 1862-1870. ISSN 0009-9147
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Publisher's URL: http://dx.doi.org/10.1373/clinchem.2010.145581
BACKGROUND: The nucleoside analogue cytarabine (Ara-C) is the key agent for treating acute myeloid leukemia (AML), however up to 30% of patients fail to respond to treatment. Screening of patient blood samples to determine drug response prior to commencement of treatment is needed. This project aimed to construct and evaluate a self-bioluminescent reporter strain of Escherichia coli for use as an Ara-C biosensor and to design an in-vitro assay to predict Ara-C response in clinical samples. METHODS: Transposition mutagenesis was used to create a cytidine deaminase (cdd)-deficient mutant of E. coli MG1655 that responded to Ara-C. The strain was transformed with the luxCDABE operon and used as a whole-cell biosensor for development of an 8-hour assay to determine Ara-C uptake and phosphorylation by leukemic cells. RESULTS: Intracellular concentrations of 0.025 µmol/L of phosphorylated Ara-C were detected by significantly increased light output (P<0.05) from the bacterial biosensor. Results using AML cell lines with known response to Ara-C, show close correlation between the 8-hour assay and a 3-day cytotoxicity test for Ara-C cell killing. In retrospective tests with 24 clinical samples of bone marrow or peripheral blood, the biosensor-based assay predicted leukemic cell response to Ara-C within 8 hours. CONCLUSIONS: The biosensor-based assay may offer a predictor for evaluating the sensitivity of leukemic cells to Ara-C before patients undergo chemotherapy and allow customized treatment of drug sensitive patients with reduced Ara-C dose levels. The 8-hour assay monitors intracellular Ara-CTP levels and if fully validated, may be suitable for use in clinical settings.
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