Metabolome profiling by HRMAS NMR spectroscopy of pheochromocytomas and paragangliomas detects SDH deficiency: clinical and pathophysiological implications

Alessio Imperiale; François-Marie Moussallieh; Philippe Roche; Stéphanie Battini; A Ercument Cicek; Frédéric Sebag; Laurent Brunaud; Anne Barlier; Karim Elbayed; Anderson Loundou; Philippe Bachellier; Bernard Goichot; Constantine A Stratakis; Karel Pacak; Izzie-Jacques Namer; David Taïeb

doi:10.1016/j.neo.2014.10.010

Metabolome profiling by HRMAS NMR spectroscopy of pheochromocytomas and paragangliomas detects SDH deficiency: clinical and pathophysiological implications

Neoplasia. 2015 Jan;17(1):55-65. doi: 10.1016/j.neo.2014.10.010.

Authors

Alessio Imperiale¹, François-Marie Moussallieh², Philippe Roche³, Stéphanie Battini⁴, A Ercument Cicek⁵, Frédéric Sebag⁶, Laurent Brunaud⁷, Anne Barlier⁸, Karim Elbayed², Anderson Loundou⁹, Philippe Bachellier¹⁰, Bernard Goichot¹¹, Constantine A Stratakis¹², Karel Pacak¹³, Izzie-Jacques Namer⁴, David Taïeb¹⁴

Affiliations

¹ Department of Biophysics and Nuclear Medicine, Hautepierre Hospital, University Hospitals of Strasbourg, Strasbourg, France; ICube, UMR 7357, University of Strasbourg/CNRS and FMTS, Faculty of Medicine, Strasbourg, France. Electronic address: alessio.imperiale@chru-strasbourg.fr.
² ICube, UMR 7357, University of Strasbourg/CNRS and FMTS, Faculty of Medicine, Strasbourg, France.
³ Integrative Structural and Chemical Biology (iSCB) and INT-3D Molecular Modeling Platform, Cancer Research Centre of Marseille, CNRS UMR7258, INSERM U1068, Institut Paoli Calmettes, Aix-Marseille University UM105, Marseille, France.
⁴ Department of Biophysics and Nuclear Medicine, Hautepierre Hospital, University Hospitals of Strasbourg, Strasbourg, France; ICube, UMR 7357, University of Strasbourg/CNRS and FMTS, Faculty of Medicine, Strasbourg, France.
⁵ Lane Center for Computational Biology, School of Computer Science, Carnegie Mellon University, Pittsburgh, PA, USA.
⁶ Department of Endocrine Surgery, La Timone University Hospital, Aix-Marseille University, Marseille, France.
⁷ Department of Digestive, Hepato-Biliary and Endocrine Surgery, Brabois University Hospital, Nancy, France.
⁸ Laboratory of Biochemistry and Molecular Biology, Conception Hospital, Aix-Marseille University, Marseille, France.
⁹ Department of Public Health, Aix-Marseille University, Marseille, France.
¹⁰ Department of Visceral Surgery and Transplantation, Hautepierre Hospital, University Hospitals of Strasbourg, Strasbourg, France.
¹¹ Department of Internal Medicine, Diabetes and Metabolic Disorders, Hautepierre Hospital, University Hospitals of Strasbourg, Strasbourg, France.
¹² Section on Genetics and Endocrinology (SEGEN), Program on Developmental Endocrinology and Genetics (PDEGEN), Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
¹³ Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
¹⁴ La Timone University Hospital, European Center for Research in Medical Imaging, Aix-Marseille University, Marseille, France.

Abstract

Succinate dehydrogenase gene (SDHx) mutations increase susceptibility to develop pheochromocytomas/paragangliomas (PHEOs/PGLs). In the present study, we evaluate the performance and clinical applications of (1)H high-resolution magic angle spinning (HRMAS) nuclear magnetic resonance (NMR) spectroscopy-based global metabolomic profiling in a large series of PHEOs/PGLs of different genetic backgrounds. Eighty-seven PHEOs/PGLs (48 sporadic/23 SDHx/7 von Hippel-Lindau/5 REarranged during Transfection/3 neurofibromatosis type 1/1 hypoxia-inducible factor 2α), one SDHD variant of unknown significance, and two Carney triad (CTr)-related tumors were analyzed by HRMAS-NMR spectroscopy. Compared to sporadic, SDHx-related PHEOs/PGLs exhibit a specific metabolic signature characterized by increased levels of succinate (P < .0001), methionine (P = .002), glutamine (P = .002), and myoinositol (P < .0007) and decreased levels of glutamate (P < .0007), regardless of their location and catecholamine levels. Uniquely, ATP/ascorbate/glutathione was found to be associated with the secretory phenotype of PHEOs/PGLs, regardless of their genotype (P < .0007). The use of succinate as a single screening test retained excellent accuracy in distinguishing SDHx versus non-SDHx-related tumors (sensitivity/specificity: 100/100%). Moreover, the quantification of succinate could be considered a diagnostic alternative for assessing SDHx-related mutations of unknown pathogenicity. We were also able, for the first time, to uncover an SDH-like pattern in the two CTr-related PGLs. The present study demonstrates that HRMAS-NMR provides important information for SDHx-related PHEO/PGL characterization. Besides the high succinate-low glutamate hallmark, SDHx tumors also exhibit high values of methionine, a finding consistent with the hypermethylation pattern of these tumors. We also found important levels of glutamine, suggesting that glutamine metabolism might be involved in the pathogenesis of SDHx-related PHEOs/PGLs.

MeSH terms

Female
Humans
Immunohistochemistry
Male
Metabolic Networks and Pathways
Metabolome*
Metabolomics* / methods
Middle Aged
Models, Molecular
Mutation
Nuclear Magnetic Resonance, Biomolecular
Paraganglioma / diagnosis
Paraganglioma / genetics*
Paraganglioma / metabolism*
Pheochromocytoma / diagnosis
Pheochromocytoma / genetics*
Pheochromocytoma / metabolism*
Positron-Emission Tomography
Protein Structure, Secondary
ROC Curve
Sensitivity and Specificity
Succinate Dehydrogenase / chemistry
Succinate Dehydrogenase / deficiency
Succinate Dehydrogenase / genetics*
Succinate Dehydrogenase / metabolism*
Succinic Acid / chemistry
Succinic Acid / metabolism
Tomography, X-Ray Computed

Substances

Succinic Acid
Succinate Dehydrogenase