title: Metabolomic profiling of drug responses in acute myeloid leukaemia cell lines.
abstract: Combined bezafibrate (BEZ) and medroxyprogesterone acetate (MPA) exert unexpected antileukaemic activities against acute myeloid leukaemia (AML) and these activities are associated with the generation of reactive oxygen species (ROS) within the tumor cells. Although the generation of ROS by these drugs is supported by preceding studies including our own, the interrelationship between the cellular effects of the drugs and ROS generation is not well understood. Here we report the use of NMR metabolomic profiling to further study the effect of BEZ and MPA on three AML cell lines and to shed light on the underlying mechanism of action. For this we focused on drug effects induced during the initial 24 hours of treatment prior to the onset of overt cellular responses and examined these in the context of basal differences in metabolic profiles between the cell lines. Despite their ultimately profound cellular effects, the early changes in metabolic profiles engendered by these drugs were less pronounced than the constitutive metabolic differences between cell types. Nonetheless, drug treatments engendered common metabolic changes, most markedly in the response to the combination of BEZ and MPA. These responses included changes to TCA cycle intermediates consistent with recently identified chemical actions of ROS. Notable amongst these was the conversion of alpha-ketoglutarate to succinate which was recapitulated by the treatment of cell extracts with exogenous hydrogen peroxide. These findings indicate that the actions of combined BEZ and MPA against AML cells are indeed mediated downstream of the generation of ROS rather than some hitherto unsuspected mechanism. Moreover, our findings demonstrate that metabolite profiles represent highly sensitive markers for genomic differences between cells and their responses to external stimuli. This opens new perspectives to use metabolic profiling as a tool to study the rational redeployment of drugs in new disease settings.
authors.affiliation: CR UK Institute for Cancer Studies, University of Birmingham, Henry Wellcome Building for Biomolecular NMR Spectroscopy (HWB-NMR), Birmingham, United Kingdom.
authors.forename: Stefano
authors.initials: S
authors.lastname: Tiziani
authors.forename: Alessia
authors.initials: A
authors.lastname: Lodi
authors.forename: Farhat L
authors.initials: FL
authors.lastname: Khanim
authors.forename: Mark R
authors.initials: MR
authors.lastname: Viant
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Bunce
authors.forename: Ulrich L
authors.initials: UL
authors.lastname: Günther
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics / Q000379:methods*; D010100:Oxygen / Q000378:metabolism; D010641:Phenotype; D017382:Reactive Oxygen Species; D013997:Time Factors
chemical_list: D000970:Antineoplastic Agents; D017382:Reactive Oxygen Species; D010100:Oxygen