Habib M. Alloush
A bioluminescent microbial biosensor for in vitro pretreatment assessment of cytarabine efficacy in leukemia
Alloush, Habib M.; Martin, Ashley D.; Ruddock, Mark W.; Angell, Johanna E.; Hill, Phil J.; Salisbury, Vyv C.; Lamont, John; Anderson, Elizabeth; Smith, M. Ann; Smith, J. Graham; Mehta, Priyanka
Authors
Ashley D. Martin
Mark W. Ruddock
Johanna E. Angell
Phil J. Hill
Vyv C. Salisbury
John Lamont
Elizabeth Anderson Elizabeth3.Anderson@uwe.ac.uk
Senior Lecturer in Haematology
M. Ann Smith
J. Graham Smith
Priyanka Mehta
Abstract
BACKGROUND: The nucleoside analog cytarabine (Ara-C [cytosine arabinoside]) 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 before 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: Weused transposition mutagenesis 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 an 8-h assay to determine Ara-C uptake and phosphorylation by leukemic cells. RESULTS: Intracellular concentrations of 0.025 μmol/L 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 showed close correlation between the 8-h 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 h. 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-h assay monitors intracellular Ara-CTP (cytosine arabinoside triphosphate) levels and, if fully validated, may be suitable for use in clinical settings. © 2010 American Association for Clinical Chemistry.
Citation
Salisbury, V. C., Hill, P. J., Angell, J. E., Ruddock, M. W., Martin, A. D., Alloush, H. M., …Mehta, P. (2010). A bioluminescent microbial biosensor for in vitro pretreatment assessment of cytarabine efficacy in leukemia. Clinical Chemistry, 56(12), 1862-1870. https://doi.org/10.1373/clinchem.2010.145581
Journal Article Type | Article |
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Publication Date | Dec 1, 2010 |
Deposit Date | Jun 13, 2011 |
Publicly Available Date | Feb 10, 2016 |
Journal | Clinical Chemistry |
Print ISSN | 0009-9147 |
Electronic ISSN | 1530-8561 |
Publisher | American Association for Clinical Chemistry |
Peer Reviewed | Peer Reviewed |
Volume | 56 |
Issue | 12 |
Pages | 1862-1870 |
DOI | https://doi.org/10.1373/clinchem.2010.145581 |
Keywords | biolumuinescent, microbial, biosensor, in vitro, pretreatment, cytarabine, Leukemia |
Public URL | https://uwe-repository.worktribe.com/output/972940 |
Publisher URL | http://dx.doi.org/10.1373/clinchem.2010.145581 |
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