Lucie Evenepoel1,2,3, Francien H van Nederveen4, Lindsey Oudijk2, Thomas G Papathomas2,5, David F Restuccia2, Eric J T Belt6, Wouter W de Herder7, Richard A Feelders7, Gaston J H Franssen6, Marc Hamoir8, Dominique Maiter9, Hans K Ghayee10, Jerry W Shay11, Aurel Perren12, Henri J L M Timmers13, Susanne van Eeden14, Laurent Vroonen15, Selda Aydin16, Mercedes Robledo17,18, Miikka Vikkula3, Ronald R de Krijger2,19, Winand N M Dinjens2, Alexandre Persu1,20, Esther Korpershoek2. 1. Pole of Cardiovascular Research, Institut de Recherche Expérimentale et Clinique, Université catholique de Louvain, Brussels, Belgium. 2. Department of Pathology, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, Netherlands. 3. Human Molecular Genetics, de Duve Institute, Université catholique de Louvain, Brussels, Belgium. 4. Laboratory for Pathology, PAL Dordrecht, Dordrecht, Netherlands. 5. Department of Histopathology, King's College Hospital, London, United Kingdom. 6. Department of Surgery, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, Netherlands. 7. Internal Medicine, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, Netherlands. 8. Otolaryngology Department, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium. 9. Endocrinology Department, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium. 10. Department of Internal Medicine, Division of Endocrinology, University of Florida, Gainesville, Florida. 11. Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas. 12. Clinical Pathology Division, University of Bern, Bern, Switzerland. 13. Department of Internal Medicine, Division of Endocrinology, Radboud University Medical Centre, Nijmegen, Netherlands. 14. Department of Pathology, Academic Medical Center, Amsterdam, Netherlands. 15. Department of Endocrinology, Centre Hospitalier Universitaire de Liège, University of Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium. 16. Department of Pathology, Cliniques universitaires Saint Luc, Université catholique de Louvain, Brussels, Belgium. 17. Hereditary Endocrine Cancer Group, Human Cancer Genetics Programme, Spanish National Cancer Research Centre, Madrid, Spain. 18. Centre for Biomedical Network Research on Rare Diseases, Madrid, Spain. 19. Department of Pathology, Reinier de Graaf Hospital, Delft, Netherlands. 20. Division of Cardiology, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium.
Abstract
Context: Pheochromocytomas and paragangliomas (PPGLs) are rare neuroendocrine, usually benign, tumors. Currently, the only reliable criterion of malignancy is the presence of metastases. Objective: The aim of this study was to identify genes associated with malignancy in PPGLs. Design: Transcriptomic profiling was performed on 40 benign and 11 malignant PPGLs. Genes showing a significantly different expression between benign and malignant PPGLs with a ratio ≥4 were confirmed and tested in an independent series by quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemistry was performed for the validated genes on 109 benign and 32 malignant PPGLs. Functional assays were performed with hPheo1 cells. Setting: This study was conducted at the Department of Pathology of the Erasmus MC University Medical Center Rotterdam Human Molecular Genetics laboratory of the de Duve Institute, University of Louvain. Patients: PPGL samples from 179 patients, diagnosed between 1972 and 2015, were included. Main outcome measures: Associations between gene expression and malignancy were tested using supervised clustering approaches. Results: Ten differentially expressed genes were selected based on messenger RNA (mRNA) expression array data. Contactin 4 (CNTN4) was overexpressed in malignant vs benign tumors [4.62-fold; false discovery rate (FDR), 0.001]. Overexpression at the mRNA level was confirmed using qRT-PCR (2.90-fold, P = 0.02; validation set: 4.26-fold, P = 0.005). Consistent findings were obtained in The Cancer Genome Atlas cohort (2.7-fold; FDR, 0.02). CNTN4 protein was more frequently expressed in malignant than in benign PPGLs by immunohistochemistry (58% vs 17%; P = 0.002). Survival after 7 days of culture under starvation conditions was significantly enhanced in hPheo1 cells transfected with CNTN4 complementary DNA. Conclusion: CNTN4 expression is consistently associated with malignant behavior in PPGLs.
Context:Pheochromocytomas and paragangliomas (PPGLs) are rare neuroendocrine, usually benign, tumors. Currently, the only reliable criterion of malignancy is the presence of metastases. Objective: The aim of this study was to identify genes associated with malignancy in PPGLs. Design: Transcriptomic profiling was performed on 40 benign and 11 malignant PPGLs. Genes showing a significantly different expression between benign and malignant PPGLs with a ratio ≥4 were confirmed and tested in an independent series by quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemistry was performed for the validated genes on 109 benign and 32 malignant PPGLs. Functional assays were performed with hPheo1 cells. Setting: This study was conducted at the Department of Pathology of the Erasmus MC University Medical Center Rotterdam Human Molecular Genetics laboratory of the de Duve Institute, University of Louvain. Patients: PPGL samples from 179 patients, diagnosed between 1972 and 2015, were included. Main outcome measures: Associations between gene expression and malignancy were tested using supervised clustering approaches. Results: Ten differentially expressed genes were selected based on messenger RNA (mRNA) expression array data. Contactin 4 (CNTN4) was overexpressed in malignant vs benign tumors [4.62-fold; false discovery rate (FDR), 0.001]. Overexpression at the mRNA level was confirmed using qRT-PCR (2.90-fold, P = 0.02; validation set: 4.26-fold, P = 0.005). Consistent findings were obtained in The Cancer Genome Atlas cohort (2.7-fold; FDR, 0.02). CNTN4 protein was more frequently expressed in malignant than in benign PPGLs by immunohistochemistry (58% vs 17%; P = 0.002). Survival after 7 days of culture under starvation conditions was significantly enhanced in hPheo1 cells transfected with CNTN4 complementary DNA. Conclusion:CNTN4 expression is consistently associated with malignant behavior in PPGLs.
Authors: Magnus Zethoven; Luciano Martelotto; Andrew Pattison; Blake Bowen; Shiva Balachander; Aidan Flynn; Fernando J Rossello; Annette Hogg; Julie A Miller; Zdenek Frysak; Sean Grimmond; Lauren Fishbein; Arthur S Tischler; Anthony J Gill; Rodney J Hicks; Patricia L M Dahia; Roderick Clifton-Bligh; Karel Pacak; Richard W Tothill Journal: Nat Commun Date: 2022-10-21 Impact factor: 17.694