Abstract
Pseudohypoparathyroidism is a rare endocrine disorder that can be caused by genetic (mainly maternally inherited inactivating point mutations, although intragenic and gross deletions have rarely been reported) or epigenetic alterations at GNAS locus. Clinical and molecular characterization of this disease is not that easy because of phenotypic, biochemical and molecular overlapping features between both subtypes of the disease. The European Consortium for the study of PHP (EuroPHP) designed the present work with the intention of generating the standards of diagnostic clinical molecular (epi)genetic testing in PHP patients. With this aim, DNA samples of eight independent PHP patients carrying GNAS genetic and/or epigenetic defects (three patients with GNAS deletions, two with 20q uniparental disomy and three with a methylation defect of unknown origin) without GNAS point mutations were anonymized and sent to the five participant laboratories for their routine genetic analysis (methylation-specific (MS)-MLPA, pyrosequencing and EpiTYPER) and interpretations. All laboratories were able to detect methylation defects and, after the data analysis, the Consortium compared the results to define technical advantages and disadvantages of different techniques. To conclude, we propose as first-level investigation in PHP patients copy number and methylation analysis by MS-MLPA. Then, in patients with partial methylation defect, the result should be confirmed by single CpG bisulphite-based methods (ie pyrosequencing), whereas in case of a complete methylation defect without detectable deletion, microsatellites or SNP genotyping should be performed to exclude uniparental disomy 20.
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Change history
12 March 2015
This article has been amended since online publication. A corrigendum also appears in this issue.
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Acknowledgements
This work was supported by the Euro-Pseudohypoparathyroidism network (EuroPHP) grant from the European Society for Pediatric Endocrinology Research Unit (to AL), grants from the University of Luebeck (awarded to ST), Attractivité Grant from Paris-Sud University 2013 (to AL), ANR EPIFEGROW (Agence nationale de la recherche) (to AL), French Society of Pediatric Endocrinology and Diabetology (to VG), the Italian Ministry of Health (to GM: GR-2009-1608394), recurrent funding from INSERM U986 (to AL, VG and CS) and Instituto de Salud Carlos III (PI10/0148 and PI13/00467 to GPdN). KF and BI are supported by research Grants G.0490.10N and G.0A23.14N from the FWO-Vlaanderen (Belgium); IG by FIS-Program (I3SNS-CA10/01056) and GPdN by I3SNS Program of the Spanish Ministry of Health (CP03/0064; SIVI 1395/09). All members of the EuroPHP are members of the EUCID.net (COST action BM1208 on imprinting disorders; http: //www.imprinting-disorders.eu).
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Belgium (Kathleen Freson and Benedetta Izzi, Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven), France (Agnes Linglart, Caroline Silve, Stephanie Maupetit-Mehouas and Virginie Grybek, INSERM U986, Hôpital Bicêtre, Le Kremlin Bicêtre, Paris; and Anne Barlier, Laboratory of Biochemistry and Molecular Biology, Conception Hospital, Aix-Marseille University, Marseille), Germany (Olaf Hiort, Susanne Thiele, Bettina Brix and Ralf Werner, Division of Experimental Paediatric Endocrinology and Diabetes, Department of Paediatrics, University of Luebeck), Italy (Giovanna Mantovani, Francesca M Elli, Paolo Bordogna and Valentina Boldrin, Department of Clinical Sciences and Community Health, University of Milan, Endocrinology and Diabetology Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan; and Luisa de Sanctis, Department of Public Health and Pediatrics, University of Turin, Reggina Margherita Children’s Hospital, Turin), Spain (Guiomar Perez de Nanclares, Intza Garin and Arrate Pereda, Molecular (Epi)Genetics Laboratory, BioAraba National Health Institute, Hospital Universitario Araba-Txagorritxu, Vitoria-Gasteiz; and Beatriz Lecumberri, Endocrinology Service, Hospital Universitario La Paz, Madrid), Turkey (Serap Turan, Marmara University School of Medicine Hospital, Istanbul) and United Kingdom (Deborah JG Mackay, Faculty of Medicine, University of Southampton, Southampton).
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Garin, I., Mantovani, G., Aguirre, U. et al. European guidance for the molecular diagnosis of pseudohypoparathyroidism not caused by point genetic variants at GNAS: an EQA study. Eur J Hum Genet 23, 438–444 (2015). https://doi.org/10.1038/ejhg.2014.127
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DOI: https://doi.org/10.1038/ejhg.2014.127
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