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)

Thomas G Papathomas; Lindsey Oudijk; Alexandre Persu; Anthony J Gill; Francien van Nederveen; Arthur S Tischler; Frédérique Tissier; Marco Volante; Xavier Matias-Guiu; Marcel Smid; Judith Favier; Elena Rapizzi; Rosella Libe; Maria Currás-Freixes; Selda Aydin; Thanh Huynh; Urs Lichtenauer; Anouk van Berkel; Letizia Canu; Rita Domingues; Roderick J Clifton-Bligh; Magdalena Bialas; Miikka Vikkula; Gustavo Baretton; Mauro Papotti; Gabriella Nesi; Cécile Badoual; Karel Pacak; Graeme Eisenhofer; Henri J Timmers; Felix Beuschlein; Jérôme Bertherat; Massimo Mannelli; Mercedes Robledo; Anne-Paule Gimenez-Roqueplo; Winand Nm Dinjens; Esther Korpershoek; Ronald R de Krijger

doi:10.1038/modpathol.2015.41

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 Jun;28(6):807-21. doi: 10.1038/modpathol.2015.41. Epub 2015 Feb 27.

Authors

Thomas G Papathomas¹, Lindsey Oudijk², Alexandre Persu³, Anthony J Gill⁴, Francien van Nederveen⁵, Arthur S Tischler⁶, Frédérique Tissier⁷, Marco Volante⁸, Xavier Matias-Guiu⁹, Marcel Smid¹⁰, Judith Favier¹¹, Elena Rapizzi¹², Rosella Libe¹³, Maria Currás-Freixes¹⁴, Selda Aydin¹⁵, Thanh Huynh¹⁶, Urs Lichtenauer¹⁷, Anouk van Berkel¹⁸, Letizia Canu¹², Rita Domingues¹⁹, Roderick J Clifton-Bligh²⁰, Magdalena Bialas²¹, Miikka Vikkula²², Gustavo Baretton²³, Mauro Papotti⁸, Gabriella Nesi²⁴, Cécile Badoual²⁵, Karel Pacak¹⁶, Graeme Eisenhofer²⁶, Henri J Timmers¹⁸, Felix Beuschlein¹⁷, Jérôme Bertherat²⁷, Massimo Mannelli²⁸, Mercedes Robledo²⁹, Anne-Paule Gimenez-Roqueplo¹¹, Winand Nm Dinjens², Esther Korpershoek², Ronald R de Krijger³⁰

Affiliations

¹ 1] Department of Histopathology, King's College Hospital, London, UK [2] Department of Pathology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, Rotterdam, The Netherlands.
² Department of Pathology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, Rotterdam, The Netherlands.
³ Pole of Cardiovascular Research, Institut de Recherche Expérimentale et Clinique and Division of Cardiology, Cliniques Universitaires Saint-Luc, Université Catholique de Louvain, Brussels, Belgium.
⁴ 1] Department of Anatomical Pathology, Royal North Shore Hospital, St Leonards, NSW, Australia [2] Cancer Diagnosis and Pathology Research Group, Kolling Institute of Medical Research, University of Sydney, Sydney, NSW, Australia.
⁵ Laboratory for Pathology, PAL Dordrecht, Dordrecht, The Netherlands.
⁶ Department of Pathology and Laboratory Medicine, Tufts Medical Center, Tufts University School of Medicine, Boston, MA, USA.
⁷ 1] Institut National de la Santé et de la Recherche Médicale U1016, Institut Cochin, Centre National de la Recherche Scientifique UMR8104, Université Paris Descartes, Sorbonne Paris Cité, Rare Adrenal Cancer Network COMETE, Paris, France [2] Department of Pathology, Hôpital Pitié-Salpêtrière, Université Pierre et Marie Curie, Paris, France.
⁸ Department of Oncology, University of Turin at San Luigi Hospital, Turin, Italy.
⁹ Department of Pathology and Molecular Genetics and Research Laboratory, Hospital Universitari Arnau de Vilanova, IRBLLEIDA, University of Lleida, Lleida, Spain.
¹⁰ Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, Rotterdam, The Netherlands.
¹¹ 1] Paris-Centre de Recherche Cardiovasculaire (PARCC), Inserm UMR970, Hôpital Européen Georges Pompidou, Paris, France [2] Université Paris Descartes, Faculté de Médecine, Paris Cité Sorbonne, France.
¹² Endocrinology Unit, Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy.
¹³ Institut National de la Santé et de la Recherche Médicale U1016, Institut Cochin, Centre National de la Recherche Scientifique UMR8104, Université Paris Descartes, Sorbonne Paris Cité, Rare Adrenal Cancer Network COMETE, Paris, France.
¹⁴ Hereditary Endocrine Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
¹⁵ Department of Pathology, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Institut de Recherche Expérimentale et Clinique, Brussels, Belgium.
¹⁶ Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
¹⁷ Endocrine Research Unit, Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
¹⁸ Department of Internal Medicine, Section of Endocrinology, Radboud University Medical Centre, Nijmegen, The Netherlands.
¹⁹ Unidade de Investigação em Patobiologia Molecular (UIPM), Instituto Português de Oncologia de Lisboa Francisco Gentil, Lisbon, Portugal.
²⁰ Cancer Genetics, Kolling Institute of Medical Research, Royal North Shore Hospital, University of Sydney, Sydney, NSW, Australia.
²¹ Department of Pathomorphology, Jagiellonian University Medical College, Krakow, Poland.
²² Laboratory of Human Molecular Genetics, de Duve Institute, Université Catholique de Louvain, Brussels, Belgium.
²³ Department of Pathology, Technische Universität Dresden, Dresden, Germany.
²⁴ Division of Pathological Anatomy, University of Florence, Florence, Italy.
²⁵ 1] Paris-Centre de Recherche Cardiovasculaire (PARCC), Inserm UMR970, Hôpital Européen Georges Pompidou, Paris, France [2] Université Paris Descartes, Faculté de Médecine, Paris Cité Sorbonne, France [3] Service d'Anatomie Pathologique, Hôpital Européen Georges-Pompidou, Assistance Publique Hôpitaux de Paris (AP-HP), Paris, France.
²⁶ Institute of Clinical Chemistry and Laboratory Medicine and Department of Medicine III, University Hospital, Technische Universität Dresden, Dresden, Germany.
²⁷ 1] Institut Cochin, Université Paris Descartes, INSERM U1016, CNRS UMR8104, Paris, France [2] Department of Endocrinology, Referral Center for Rare Adrenal Diseases, Assistance Publique Hôpitaux de Paris, Hôpital Cochin, Paris, France.
²⁸ 1] Endocrinology Unit, Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy [2] Istituto Toscano Tumori (ITT), Florence, Italy.
²⁹ 1] Hereditary Endocrine Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain [2] Centre for Biomedical Network Research on Rare Diseases (CIBERER), Madrid, Spain.
³⁰ 1] Department of Pathology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, Rotterdam, The Netherlands [2] Department of Pathology, Reinier de Graaf Hospital, Delft, The Netherlands [3] Department of Pathology, University Medical Center Utrecht, Princess Maxima Center for Pediatric Oncology, Utrecht, The Netherlands.

PMID: 25720320
DOI: 10.1038/modpathol.2015.41

Abstract

Despite the established role of SDHB/SDHA immunohistochemistry as a valuable tool to identify patients at risk for familial succinate dehydrogenase-related pheochromocytoma/paraganglioma syndromes, the reproducibility of the assessment methods has not as yet been determined. The aim of this study was to investigate interobserver variability among seven expert endocrine pathologists using a web-based virtual microscopy approach in a large multicenter pheochromocytoma/paraganglioma cohort (n=351): (1) 73 SDH mutated, (2) 105 non-SDH mutated, (3) 128 samples without identified SDH-x mutations, and (4) 45 with incomplete SDH molecular genetic analysis. Substantial agreement among all the reviewers was observed either with a two-tiered classification (SDHB κ=0.7338; SDHA κ=0.6707) or a three-tiered classification approach (SDHB κ=0.6543; SDHA κ=0.7516). Consensus was achieved in 315 cases (89.74%) for SDHB immunohistochemistry and in 348 cases (99.15%) for SDHA immunohistochemistry. Among the concordant cases, 62 of 69 (~90%) SDHB-/C-/D-/AF2-mutated cases displayed SDHB immunonegativity and SDHA immunopositivity, 3 of 4 (75%) with SDHA mutations showed loss of SDHA/SDHB protein expression, whereas 98 of 105 (93%) non-SDH-x-mutated counterparts demonstrated retention of SDHA/SDHB protein expression. Two SDHD-mutated extra-adrenal paragangliomas were scored as SDHB immunopositive, whereas 9 of 128 (7%) tumors without identified SDH-x mutations, 6 of 37 (~16%) VHL-mutated, as well as 1 of 21 (~5%) NF1-mutated tumors were evaluated as SDHB immunonegative. Although 14 out of those 16 SDHB-immunonegative cases were nonmetastatic, an overall significant correlation between SDHB immunonegativity and malignancy was observed (P=0.00019). We conclude that SDHB/SDHA immunohistochemistry is a reliable tool to identify patients with SDH-x mutations with an additional value in the assessment of genetic variants of unknown significance. If SDH molecular genetic analysis fails to detect a mutation in SDHB-immunonegative tumor, SDHC promoter methylation and/or VHL/NF1 testing with the use of targeted next-generation sequencing is advisable.

Publication types

Multicenter Study
Research Support, Non-U.S. Gov't

MeSH terms

Adrenal Gland Neoplasms / genetics*
Biomarkers, Tumor / analysis
Biomarkers, Tumor / genetics
Electron Transport Complex II / analysis*
Electron Transport Complex II / genetics
Humans
Immunohistochemistry / standards*
Microscopy / methods
Mutation
Observer Variation
Paraganglioma / genetics*
Pheochromocytoma / genetics*
Succinate Dehydrogenase / analysis*
Succinate Dehydrogenase / genetics
Telepathology / methods

Substances

Biomarkers, Tumor
Electron Transport Complex II
SDHA protein, human
SDHB protein, human
Succinate Dehydrogenase