Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line

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 li...

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Main Authors:	Carrigan, John B., Reed, Michelle A. C., Ludwig, Christian, Khanim, Farhat L., Bunce, Christopher M., Günther, Ulrich L.
Format:	Online
Language:	English
Published:	John Wiley and Sons Inc. 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4916734/

id	pubmed-4916734
recordtype	oai_dc
spelling	pubmed-49167342016-06-22 Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line Carrigan, John B. Reed, Michelle A. C. Ludwig, Christian Khanim, Farhat L. Bunce, Christopher M. Günther, Ulrich L. Full Papers 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 13C‐tracer‐based NMR metabolic study to understand how these drugs work in K562 leukaemia cells. Our study shows that [1,2‐13C]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 [3‐13C]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. John Wiley and Sons Inc. 2016-03-22 2016-05 /pmc/articles/PMC4916734/ /pubmed/27347458 http://dx.doi.org/10.1002/cplu.201500549 Text en © 2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Carrigan, John B. Reed, Michelle A. C. Ludwig, Christian Khanim, Farhat L. Bunce, Christopher M. Günther, Ulrich L.
spellingShingle	Carrigan, John B. Reed, Michelle A. C. Ludwig, Christian Khanim, Farhat L. Bunce, Christopher M. Günther, Ulrich L. Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
author_facet	Carrigan, John B. Reed, Michelle A. C. Ludwig, Christian Khanim, Farhat L. Bunce, Christopher M. Günther, Ulrich L.
author_sort	Carrigan, John B.
title	Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
title_short	Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
title_full	Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
title_fullStr	Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
title_full_unstemmed	Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line
title_sort	tracer‐based metabolic nmr‐based flux analysis in a leukaemia cell line
description	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 13C‐tracer‐based NMR metabolic study to understand how these drugs work in K562 leukaemia cells. Our study shows that [1,2‐13C]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 [3‐13C]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.
publisher	John Wiley and Sons Inc.
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4916734/
_version_	1613598083944284160

Tracer‐Based Metabolic NMR‐Based Flux Analysis in a Leukaemia Cell Line

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