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Conditions for C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas
Authors:Kumar Pichumani  Tomoyuki Mashimo  Hyeon-Man Baek  James Ratnakar  Bruce Mickey  Ralph J. DeBerardinis  Elizabeth A. Maher  Robert M. Bachoo  Craig R. Malloy  Zoltan Kovacs
Affiliation:1. Advanced Imaging Research Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;2. Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;3. Annette G. Strauss Center for Neuro-Oncology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;4. Center for Magnetic Resonance Research, Korea Basic Science Institute, Chungbuk 363-883, Republic of Korea;5. Department of Neurological Surgery, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;6. Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;g McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;h Children’s Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;i Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;j Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;k Department of Radiology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA;l Veterans Affairs North Texas HealthCare System, Lancaster, TX 75216, USA
Abstract:13C NMR (nuclear magnetic resonance) spectroscopy of extracts from patient tumor samples provides rich information about metabolism. However, in isocitrate dehydrogenase (IDH)-mutant gliomas, 13C labeling is obscured in oncometabolite 2-hydroxyglutaric acid (2HG) by glutamate and glutamine, prompting development of a simple method to resolve the metabolites. J-coupled multiplets in 2HG were similar to glutamate and glutamine and could be clearly resolved at pH 6. A cryogenically cooled 13C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of 13C–13C spin–spin couplings in 2HG expressing IDH-mutant gliomas.
Keywords:2-Hydroxyglutarate   Cryoprobe   13C NMR   IDH-mutant gliomas
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