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Assessment of the Metabolic Profile of Primary Leukemia Cells
Published on: November 21, 2018
Tracer-Based Metabolic NMR-Based Flux Analysis in a Leukaemia Cell Line
John B Carrigan1, Michelle A C Reed1, Christian Ludwig2
1Institute of Cancer and Genomics Sciences, University of Birmingham, Birmingham, B15 2TT, UK.
Abstract:
High levels of reactive oxygen species (ROS) have a profound impact on acute myeloid leukaemia cells and can be used to specifically target these cells with novel therapies. We have previously shown how the combination of two redeployed drugs, the contraceptive steroid medroxyprogesterone and the lipid-regulating drug bezafibrate exert anti-leukaemic effects by producing ROS. Here we report a 13 C-tracer-based NMR metabolic study to understand how these drugs work in K562 leukaemia cells. Our study shows that [1,2-13 C]glucose is incorporated into ribose sugars, indicating activity in oxidative and non-oxidative pentose phosphate pathways alongside lactate production. There is little label incorporation into the tricarboxylic acid cycle from glucose, but much greater incorporation arises from the use of [3-13 C]glutamine. The combined medroxyprogesterone and bezafibrate treatment decreases label incorporation from both glucose and glutamine into α-ketoglutarate and increased that for succinate, which is consistent with ROS-mediated conversion of α-ketoglutarate to succinate. Most interestingly, this combined treatment drastically reduced the production of several pyrimidine synthesis intermediates.
Insights
Reactive oxygen species (ROS) target acute myeloid leukaemia. Medroxyprogesterone and bezafibrate combination therapy generates ROS, impacting K562 cells
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- High reactive oxygen species (ROS) levels are implicated in acute myeloid leukaemia (AML) pathogenesis.
- Novel therapies targeting AML cells via ROS production are under investigation.
- Previous studies demonstrated anti-leukaemic effects of medroxyprogesterone and bezafibrate by inducing ROS.
Purpose of the Study:
- To elucidate the metabolic mechanisms underlying the anti-leukaemic effects of combined medroxyprogesterone and bezafibrate.
- To investigate drug-induced metabolic alterations in K562 leukaemia cells using a 13C-tracer-based NMR approach.
Main Methods:
- Utilized 13C-tracer-based Nuclear Magnetic Resonance (NMR) spectroscopy.
- Traced the metabolic fate of [1,2-13C]glucose and [3-13C]glutamine in K562 cells.
- Analyzed metabolic flux through the pentose phosphate pathway, glycolysis, and the tricarboxylic acid (TCA) cycle.
Main Results:
- Combined medroxyprogesterone and bezafibrate treatment altered glucose and glutamine metabolism.
- Decreased incorporation of labels into α-ketoglutarate and increased incorporation into succinate, suggesting ROS-mediated α-ketoglutarate to succinate conversion.
- Significant reduction in pyrimidine synthesis intermediates observed under combined drug treatment.
Conclusions:
- The study reveals distinct metabolic reprogramming in K562 cells upon combined medroxyprogesterone and bezafibrate treatment.
- Metabolic alterations, including TCA cycle modifications and suppressed pyrimidine synthesis, are linked to ROS production and contribute to the anti-leukaemic effect.
- This work provides insights into the metabolic vulnerabilities of AML cells targeted by repurposed drugs.
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