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Human melanoma metabolic network analysis with combined 13C NMR/bioreactor technique: testing the Warburg effect
Author(s) -
Shestov Alexander,
Mancuso Anthony,
Locasale Jason W,
Glickson Jerry D
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.798.1
Subject(s) - warburg effect , anaerobic glycolysis , oxidative phosphorylation , glycolysis , bioreactor , metabolic flux analysis , flux (metallurgy) , metabolic pathway , chemistry , biochemistry , metabolism , citric acid cycle , biophysics , biology , organic chemistry
The bioreactor techniques are becoming an important tool to study cancer cell metabolism(1). The goal of the present work was to develop a 13 C metabolic “bonded cumomer” modeling approach adapted for fitting bioreactor data obtained with 13 C glucose as substrate and to calculate the extent of the Warburg effect and how much energy comes from aerobic glycolysis vs oxidative phosphorylation. Results Figure 1 shows experimental time courses for labeled glutamate obtained during [1,6‐ 13 C 2 ] glucose perfusion of melanoma cells cultured in a CST bioreactor. Using extracted fluxes we estimated that 54% of the energy coming from oxidative phosphorylation (from which 44% are from glucose) and 46% from aerobic glycolysis and the extent of the Warburg effect (aerobic glycolysis) was 87%. Conclusions Under aerobic conditions glycolysis provides nearly as much ATP as oxidative phosphorylation. The model is validated by excellent agreement between model predicted and experimentally measured values of CMRO 2 and cytosolic glutamate pool size. Dynamic high‐resolution MR spectra is very sensitive to changing/adding biochemical pathways and flux values, and bonded cumomer modeling allows one to check precisely the feasibility of assumed general bionetworks and particular metabolic pathways. This work was supported by NIH grants 2U24‐CA083105 and 5R01CA129544–02