Sudhir Kumar Rai1, Fernando Bril2, Heather M Hatch3, Yiling Xu1, Laura Shelton4, Srilaxmi Kalavalapalli1, Arielle Click5, Douglas Lee6, Chris Beecher7, Austin Kirby8, Kimi Kong8, Jose Trevino9, Abhishek Jha10, Shashank Jatav10, Kriti Kriti10, Soumya Luthra10, Timothy J Garrett11, Joy Guingab-Cagmat11, Daniel Plant12, Prodip Bose12, Kenneth Cusi2, Robert A Hromas8, Arthur S Tischler13, James F Powers13, Priyanka Gupta14, James Bibb14, Felix Beuschlein15, Mercedes Robledo16, Bruna Calsina16, Henri Timmers17, David Taieb18, Matthias Kroiss19, Susan Richter20, Katharina Langton21, Graeme Eisenhofer22, Raymond Bergeron23, Karel Pacak24, Sergei G Tevosian25, Hans K Ghayee26. 1. Department of Medicine, Division of Endocrinology, University of Florida, Gainesville, FL, USA. 2. Department of Medicine, Division of Endocrinology, University of Florida and Malcom Randall VA Medical Center, Gainesville, FL, USA. 3. Department of Physiological Sciences, University of Florida, Gainesville, FL, USA. 4. Scientific Project Development, Human Metabolome Technologies, Boston, MA, USA. 5. Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX, USA. 6. Omic Insight, LLC, Durham, NC, USA. 7. IROA Technologies, Chapel Hill, NC, USA. 8. Department of Medicine, University of Texas Health Science Center, San Antonio, TX, USA. 9. Department of Surgery, University of Florida, Gainesville, FL, USA. 10. Elucidata, Cambridge, MA, USA. 11. Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, Gainesville, FL, USA. 12. Department of Physiological Sciences, University of Florida, Malcom Randall VA Medical Center, Gainesville, FL, USA. 13. Department of Pathology and Laboratory Medicine, Tufts Medical Center, Boston, MA, USA. 14. Department of Surgery, University of Alabama, Birmingham, AL, USA. 15. Klinik für Endokrinologie, Diabetologie und Klinische Ernährung, UniversitätsSpital Zurich, Zurich, Switzerland. 16. Hereditary Endocrine Cancer Group, Spanish National Cancer Research Center (CNIO), and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Madrid, Spain. 17. Department of Internal Medicine, Radboud University Medical Center, Nijmegen, Netherlands. 18. Department of Nuclear Medicine, La Timone University Hospital, European Center for Research in Medical Imaging, Aix Marseille Université, Marseille, France. 19. Department of Internal Medicine, Division of Endocrinology and Diabetology, University Hospital Würzburg, University of Würzburg, Würzburg, Germany. 20. Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany. 21. Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany. 22. Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany; Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany; Division of Clinical Neurochemistry, Institute of Clinical Chemistry and Laboratory Medicine, and Department of Medicine, Faculty of Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany. 23. Department of Medicinal Chemistry, University of Florida, Gainesville, FL, USA. 24. Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA. 25. Department of Physiological Sciences, University of Florida, Gainesville, FL, USA. Electronic address: stevosian@ufl.edu. 26. Department of Medicine, Division of Endocrinology, University of Florida and Malcom Randall VA Medical Center, Gainesville, FL, USA. Electronic address: hans.ghayee@medicine.ufl.edu.
Abstract
BACKGROUND: Pheochromocytomas (PCCs) and paragangliomas (PGLs) are neuroendocrine tumors that are mostly benign. Metastatic disease does occur in about 10% of cases of PCC and up to 25% of PGL, and for these patients no effective therapies are available. Patients with mutations in the succinate dehydrogenase subunit B (SDHB) gene tend to have metastatic disease. We hypothesized that a down-regulation in the active succinate dehydrogenase B subunit should result in notable changes in cellular metabolic profile and could present a vulnerability point for successful pharmacological targeting. METHODS: Metabolomic analysis was performed on human hPheo1 cells and shRNA SDHB knockdown hPheo1 (hPheo1 SDHB KD) cells. Additional analysis of 115 human fresh frozen samples was conducted. In vitro studies using N1,N11-diethylnorspermine (DENSPM) and N1,N12- diethylspermine (DESPM) treatments were carried out. DENSPM efficacy was assessed in human cell line derived mouse xenografts. RESULTS: Components of the polyamine pathway were elevated in hPheo1 SDHB KD cells compared to wild-type cells. A similar observation was noted in SDHx PCC/PGLs tissues compared to their non-mutated counterparts. Specifically, spermidine, and spermine were significantly elevated in SDHx-mutated PCC/PGLs, with a similar trend in hPheo1 SDHB KD cells. Polyamine pathway inhibitors DENSPM and DESPM effectively inhibited growth of hPheo1 cells in vitro as well in mouse xenografts. CONCLUSIONS: This study demonstrates overactive polyamine pathway in PCC/PGL with SDHB mutations. Treatment with polyamine pathway inhibitors significantly inhibited hPheo1 cell growth and led to growth suppression in xenograft mice treated with DENSPM. These studies strongly implicate the polyamine pathway in PCC/PGL pathophysiology and provide new foundation for exploring the role for polyamine analogue inhibitors in treating metastatic PCC/PGL. PRéCIS: Cell line metabolomics on hPheo1 cells and PCC/PGL tumor tissue indicate that the polyamine pathway is activated. Polyamine inhibitors in vitro and in vivo demonstrate that polyamine inhibitors are promising for malignant PCC/PGL treatment. However, further research is warranted.
BACKGROUND: Pheochromocytomas (PCCs) and paragangliomas (PGLs) are neuroendocrine tumors that are mostly benign. Metastatic disease does occur in about 10% of cases of PCC and up to 25% of PGL, and for these patients no effective therapies are available. Patients with mutations in the succinate dehydrogenase subunit B (SDHB) gene tend to have metastatic disease. We hypothesized that a down-regulation in the active succinate dehydrogenase B subunit should result in notable changes in cellular metabolic profile and could present a vulnerability point for successful pharmacological targeting. METHODS: Metabolomic analysis was performed on human hPheo1 cells and shRNA SDHB knockdown hPheo1 (hPheo1 SDHB KD) cells. Additional analysis of 115 human fresh frozen samples was conducted. In vitro studies using N1,N11-diethylnorspermine (DENSPM) and N1,N12- diethylspermine (DESPM) treatments were carried out. DENSPM efficacy was assessed in human cell line derived mouse xenografts. RESULTS: Components of the polyamine pathway were elevated in hPheo1 SDHB KD cells compared to wild-type cells. A similar observation was noted in SDHx PCC/PGLs tissues compared to their non-mutated counterparts. Specifically, spermidine, and spermine were significantly elevated in SDHx-mutated PCC/PGLs, with a similar trend in hPheo1 SDHB KD cells. Polyamine pathway inhibitors DENSPM and DESPM effectively inhibited growth of hPheo1 cells in vitro as well in mouse xenografts. CONCLUSIONS: This study demonstrates overactive polyamine pathway in PCC/PGL with SDHB mutations. Treatment with polyamine pathway inhibitors significantly inhibited hPheo1 cell growth and led to growth suppression in xenograft mice treated with DENSPM. These studies strongly implicate the polyamine pathway in PCC/PGL pathophysiology and provide new foundation for exploring the role for polyamine analogue inhibitors in treating metastatic PCC/PGL. PRéCIS: Cell line metabolomics on hPheo1 cells and PCC/PGL tumor tissue indicate that the polyamine pathway is activated. Polyamine inhibitors in vitro and in vivo demonstrate that polyamine inhibitors are promising for malignant PCC/PGL treatment. However, further research is warranted.
Authors: Kıvanç Birsoy; Tim Wang; Walter W Chen; Elizaveta Freinkman; Monther Abu-Remaileh; David M Sabatini Journal: Cell Date: 2015-07-30 Impact factor: 41.582