Anion-exchange chromatography mass spectrometry provides extensive coverage of primary metabolic pathways revealing altered metabolism in IDH1 mutant cells

Abstract

Altered central carbon metabolism is a hallmark of many diseases including diabetes, obesity, heart disease and cancer. Identifying metabolic changes will open opportunities for better understanding aetiological processes and identifying new diagnostic, prognostic, and therapeutic targets. Comprehensive and robust analysis of primary metabolic pathways in cells, tissues and bio-fluids, remains technically challenging. We report on the development and validation of a highly reproducible and robust untargeted method using anion-exchange tandem mass spectrometry (IC-MS) that enables analysis of 431 metabolites, providing detailed coverage of central carbon metabolism. We apply the method in an untargeted, discovery-driven workflow to investigate the metabolic effects of isocitrate dehydrogenase 1 (IDH1) mutations in glioblastoma cells. IC-MS provides comprehensive coverage of central metabolic pathways revealing significant elevation of 2-hydroxyglutarate and depletion of 2-oxoglutarate. Further analysis of the data reveals depletion in additional metabolites including previously unrecognised changes in lysine and tryptophan metabolism.

A Novel Glycolysis-Related Four-mRNA Signature for Predicting the Survival of Patients With Breast Cancer

“…Central carbon metabolism includes the carbon utilization pathways of glycolysis, the citrate cycle, and the pentose phosphate pathway. Many diseases have changes in central carbon metabolism, including cancer, diabetes, and heart disease (Walsby-Tickle et al, 2020 ). Schraw et al ( 2019 ) showed that changes in central carbon metabolism and amino acid metabolism were related to minimal residual disease (MRD) positivity and that metabonomics has potential utility in the risk prediction and targeted treatment of pediatric acute lymphoblastic leukemia (ALL).…”

Hyperpolarized magnetic resonance shows that the anti‐ischemic drug meldonium leads to increased flux through pyruvate dehydrogenase in vivo resulting in improved post‐ischemic function in the diabetic heart

“…Terminal fed blood samples were assessed for low‐molecular‐weight metabolites with liquid chromatography‐mass spectrometry (LC‐MS) within the Department of Chemistry, University of Oxford. Plasma samples were filtered through molecular weight cut‐off filters (10 kDa) to remove proteins 25 . The infranatant was recovered and evaporated to dryness under reduced pressure.…”

Hyperpolarized Magnetic Resonance Shows that the Anti-Ischemic Drug, Meldonium, Leads to Increased Flux Through Pyruvate Dehydrogenase In Vivo Resulting in Improved Post-Ischemic Function in the Diabetic Heart.

“…Terminal fed blood samples were assessed for low molecular weight metabolites with Liquid Chromatography-Mass Spectrometry (LC-MS) within the Department of Chemistry, University of Oxford. Plasma samples were ltered through molecular weight cut-off lters (10kD) to remove proteins [25]. The infranatant was recovered and evaporated to dryness under reduced pressure.…”

“…LC-MS: Acyl-carnitines were separated and resolved using hydrophobic-interaction liquid chromatography-mass spectrometry (HILIC). Samples were separated as previously outlined [25,26]. Brie y, samples were eluted using a binary solvent, acetonitrile:water (50%:50%) containing ammonium acetate (10mM nal concentration [Solvent A]) and acetonitrile:water (95%:5%) containing ammonium acetate (10mM nal concentration [Solvent B]).…”

“…Putative compounds were identi ed with reference to authenticated standards for selected acyl-carnitines using retention time, accurate mass and fragmentation pattern to identify individual compounds [25]. Concentrations were calculated with reference to speci c standard curves.…”