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A bioluminescent microbial biosensor for in vitro pretreatment assessment of cytarabine efficacy in leukemia

Salisbury, Vyv C.; Hill, Phil J.; Angell, Johanna E.; Ruddock, Mark W.; Martin, Ashley D.; Alloush, Habib M.; Anderson, Elizabeth; Lamont, John; Smith, M. Ann; Smith, J. Graham; Mehta, Priyanka

A bioluminescent microbial biosensor for in vitro pretreatment assessment of cytarabine efficacy in leukemia Thumbnail


Authors

Vyv C. Salisbury

Phil J. Hill

Johanna E. Angell

Mark W. Ruddock

Ashley D. Martin

Habib M. Alloush

John Lamont

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.

Journal Article Type Article
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
Contract Date Feb 10, 2016

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