marmara med j Marmara Medical Journal 1019-1941 1309-9469 Marmara University 10.5472/marumj.1629043 Surgery (Other) Cerrahi (Diğer) Succinate dehydrogenase-B fluorescence in situ hybridization in pheochromocytoma-paragangliomas https://orcid.org/0000-0003-1389-3517 Keskin Öztürk Beyza The Council of Forensic Medicine https://orcid.org/0000-0003-1512-1166 Akar Kadriye Ebru The Council of Forensic Medicine, https://orcid.org/0000-0002-2009-6164 Toper Muhammed Hasan MARMARA UNIVERSITY https://orcid.org/0000-0002-6656-8227 Bagci Pelin MARMARA UNIVERSITY https://orcid.org/0000-0002-1147-4178 Kaya Handan MARMARA UNIVERSITY https://orcid.org/0000-0002-6737-6569 Cinel Leyla MARMARA UNIVERSITY 01 29 2025 38 1 76 83 10 03 2024 12 19 2024 Copyright © 1988, Marmara Medical Journal 1988 Marmara Medical Journal

Objective: Preoperative identification of familial pheochromocytoma and paraganglioma (PPGL) is crucial, yet often overlooked, leadingto missed diagnoses. Typically, succinate dehydrogenase-B (SDHB) and fumarate hydratase (FH) tests are applied postoperatively toconfirm familial links and assess prognosis. However, routine preoperative multidisciplinary collaboration is limited, causing delayedscreening requests. Consequently, routine SDHB and FH immunohistochemistry (IHC) testing is not widely practiced. This studyintroduces SDHB fluorescence in situ hybridization (FISH) as a diagnostic tool, akin to HER2 IHC-FISH testing in gastric carcinomas.Materials and Methods: Succinate dehydrogenase-B and FH IHC were conducted on 43 cases. FISH analysis was performed for 28cases with suspected familial origin or SDHB IHC loss to determine whether the protein loss was due to chromosomal changes.Results: Complete SDHB IHC loss occurred in 8 cases, partial loss in 4, and preservation in 31. Complete FH loss occurred in 10 cases.FISH analysis revealed chromosomal breaks in 20 cases (71.4%), including those with SDHB/FH IHC loss or positive clinical history.Ten cases (35.7%) showed a “red-signal only” pattern, suggesting further genetic testing.Conclusion: Succinate dehydrogenase-B FISH serves as a cost-effective tool for early PPGL diagnosis, complementing SDHB and FHIHC results. It can help identify cases that need genetic testing, even when IHC results are preserved.

 Pheochromocytoma Paraganglioma Succinate dehydrogenase-B van Nederveen FH, Gaal J, Favier J, et al. An immunohistochemical procedure to detect patients with paraganglioma and phaeochromocytoma with germline SDHB, SDHC, or SDHD gene mutations: a retrospective and prospective analysis. Lancet Oncol 2009;10:764-71. doi:10.1016/s1470-2045(09)70164-0 Mete O, Tischler AS, de Krijger R, et al. Protocol for the examination of specimens from patients with pheochromocytomas and extra-adrenal paragangliomas. Arch Pathol Lab Med 2014;138:182-8. doi:10.5858/arpa.2012- 0551-OA Zhikrivetskaya SO, Snezhkina AV, Zaretsky AR, et al. Molecular markers of paragangliomas/pheochromocytomas. Oncotarget 2017;8:25756-25782. doi:10.18632/oncotarget.15201 Cascón A, Calsina B, Monteagudo M, et al. Genetic bases of pheochromocytoma and paraganglioma. J Mol Endocrinol 2023;70:e220167. doi:10.1530/jme-22-0167 Mete O, Asa SL, Gill AJ, Kimura N, de Krijger RR, Tischler A. Overview of the 2022 WHO Classification of Paragangliomas and Pheochromocytomas. Endocr Pathol 2022;33:90-114. doi:10.1007/s12022.022.09704-6 Ricketts C, Woodward ER, Killick P, et al. Germline SDHB mutations and familial renal cell carcinoma. J Natl Cancer 2008;100:1260-2. doi:10.1093/jnci/djn254 Eisenhofer G, Tischler AS, de Krijger RR. Diagnostic tests and biomarkers for pheochromocytoma and extra-adrenal paraganglioma: from routine laboratory methods to disease stratification. Endocr Pathol 2012;23:4-14. doi:10.1007/ s12022.011.9188-1 Brouwers FM, Eisenhofer G, Tao JJ, et al. High frequency of SDHB germline mutations in patients with malignant catecholamine-producing paragangliomas: implications for genetic testing. J Clin Endocrinol Metab 2006;91:4505-9. doi:10.1210/jc.2006-0423 Su T, Yang Y, Jiang L, et al. SDHB immunohistochemistry for prognosis of pheochromocytoma and paraganglioma: A retrospective and prospective analysis. Front Endocrinol (Lausanne) 2023;14:1121397. doi:10.3389/ fendo.2023.112.1397 Koh JM, Ahn SH, Kim H, et al. Validation of pathological grading systems for predicting metastatic potential in pheochromocytoma and paraganglioma. PLoS One 2017;12:e0187398. doi:10.1371/journal.pone.0187398 Pierre C, Agopiantz M, Brunaud L, et al. COPPS, a composite score integrating pathological features, PS100 and SDHB losses, predicts the risk of metastasis and progression-free survival in pheochromocytomas/paragangliomas. Virchows Arch 2019;474:721-34. doi:10.1007/s00428.019.02553-5 Chatzikyriakou P, Brempou D, Quinn M, et al. A comprehensive characterisation of phaeochromocytoma and paraganglioma tumours through histone protein profiling, DNA methylation and transcriptomic analysis genome wide. Clin Epigenetics 2023;15:196. doi:10.1186/s13148.023.01598-3 Gottlieb E, Tomlinson IP. Mitochondrial tumour suppressors: a genetic and biochemical update. Nat Rev Cancer 2005;5:857- 66. doi:10.1038/nrc1737 Rana HQ, Koeller DR, Walker M, et al. Advancing precision oncology in hereditary paraganglioma-pheochromocytoma syndromes: Integrated interpretation and data sharing of the germline and tumor genomes. Cancers (Basel) 2024;16.947. doi:10.3390/cancers16050947 Clark GR, Sciacovelli M, Gaude E, et al. Germline FH mutations presenting with pheochromocytoma. J Clin Endocrinol Metab 2014;99:E2046-50. doi:10.1210/jc.2014-1659 Castro-Vega LJ, Buffet A, De Cubas AA, et al. Germline mutations in FH confer predisposition to malignant pheochromocytomas and paragangliomas. Hum Mol Genet 2014;23:2440-6. doi:10.1093/hmg/ddt639 Wang Y, Liu B, Li F, et al. The connection between tricarboxylic acid cycle enzyme mutations and pseudohypoxic signaling in pheochromocytoma and paraganglioma. Front Endocrinol (Lausanne) 2023;14:1274239. doi:10.3389/ fendo.2023.127.4239 Smith SC, Sirohi D, Ohe C, et al. A distinctive, low-grade oncocytic fumarate hydratase-deficient renal cell carcinoma, morphologically reminiscent of succinate dehydrogenasedeficient renal cell carcinoma. Histopathology 2017;71:42-52. doi:10.1111/his.13183 Udager AM, Magers MJ, Goerke DM, et al. The utility of SDHB and FH immunohistochemistry in patients evaluated for hereditary paraganglioma-pheochromocytoma syndromes. Hum Pathol 2018;71:47-54. doi:10.1016/j. humpath.2017.10.013 Gill AJ. Succinate dehydrogenase (SDH) and mitochondrial driven neoplasia. Pathology 2012;44:285-92. doi:10.1097/ PAT.0b013e328.353.9932 Wong MY, Andrews KA, Challis BG, et al. Clinical Practice Guidance: Surveillance for phaeochromocytoma and paraganglioma in paediatric succinate dehydrogenase gene mutation carriers. Clin Endocrinol (Oxf) 2019;90:499-505. doi:10.1111/cen.13926 Bholah R, Bunchman TE. Review of pediatric pheochromocytoma and paraganglioma. Front Pediatr 2017;5:155. doi:10.3389/fped.2017.00155 Hescot S, Curras-Freixes M, Deutschbein T, et al. Prognosis of malignant pheochromocytoma and paraganglioma (MAPP-Prono Study): A European network for the study of adrenal tumors retrospective study. J Clin Endocrinol Metab 2019;104:2367-74. doi:10.1210/jc.2018-01968 Broudin C, Favier J, Verkarre V, Méatchi T. [Pathologist contribution in the diagnosis of hereditary predisposition to paranganglioma and pheochromocytoma]. Ann Pathol 2020;40:134-41. Apport du pathologiste dans les prédispositions héréditaires aux paragangliomes et phéochromocytomes. doi:10.1016/j.annpat.2020.01.010 Gill AJ, Benn DE, Chou A, et al. Immunohistochemistry for SDHB triages genetic testing of SDHB, SDHC, and SDHD in paraganglioma-pheochromocytoma syndromes. Hum Pathol 2010;41:805-14. doi:10.1016/j.humpath.2009.12.005 Cheung VKY, Gill AJ, Chou A. Old, new, and emerging immunohistochemical markers in pheochromocytoma and paraganglioma. Endocr Pathol 2018;29:169-75. doi:10.1007/ s12022.018.9534-7 Thosani S, Ayala-Ramirez M, Palmer L, et al. The characterization of pheochromocytoma and its impact on overall survival in multiple endocrine neoplasia type 2. J Clin Endocrinol Metab 2013;98:E1813-9. doi:10.1210/jc.2013-1653 Li Q, Lan Z, Jiang Y, Wang R, Li Z, Jiang X. Validation and evaluation of 5 scoring systems for predicting metastatic risk in pheochromocytoma and paraganglioma. Am J Surg Pathol 2024;48:855-65. doi:10.1097/pas.000.000.0000002238 Papathomas TG, Oudijk L, Persu A, et al. SDHB/SDHA immunohistochemistry in pheochromocytomas and paragangliomas: a multicenter interobserver variation analysis using virtual microscopy: a multinational study of the european network for the study of adrenal tumors (ENS@T). Mod Pathol 2015;28:807-21. doi:10.1038/modpathol.2015.41 Santi R, Rapizzi E, Canu L, et al. Potential pitfalls of SDH immunohistochemical detection in paragangliomas and phaeochromocytomas harbouring germline SDHx gene mutation. Anticancer Res 2017;37:805-12. doi:10.21873/ anticanres.11381 Ohmoto A, Hayashi N, Takahashi S, Ueki A. Current prospects of hereditary adrenal tumors: towards better clinical management. Hered Cancer Clin Pract 2024;22:4. doi:10.1186/ s13053.024.00276-6 National Comprehensive Cancer Network N. Neuroendocrine and Adrenal Tumors. 2023.