Maria Rachele Ceccarini1,2, Vincenza Precone3, Elena Manara3, Stefano Paolacci4, Paolo Enrico Maltese4, Valentina Benfatti5, Kristjana Dhuli3, Kevin Donato3, Giulia Guerri4, Giuseppe Marceddu3, Pietro Chiurazzi6,7, Laura Dalla Ragione5,8, Tommaso Beccari9,10, Matteo Bertelli3,4. 1. Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy. mariarachele.ceccarini@unipg.it. 2. C.I.B., Consorzio Interuniversitario per le Biotecnologie, Trieste, Italy. mariarachele.ceccarini@unipg.it. 3. MAGI EUREGIO, Bolzano, Italy. 4. MAGI'S LAB, Rovereto, TN, Italy. 5. Department of Eating Disorder, Palazzo Francisci Todi, USL 1 Umbria, Todi, PG, Italy. 6. Dipartimento Universitario Scienze della Vita e Sanità Pubblica, Sezione di Medicina Genomica, Università Cattolica del Sacro Cuore, 00168, Rome, Italy. 7. Fondazione Policlinico Universitario "A. Gemelli" IRCCS, UOC Genetica Medica, 00168, Roma, Italy. 8. Food Science and Human Nutrition Unit, University Campus Biomedico of Rome, Rome, Italy. 9. Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy. 10. C.I.B., Consorzio Interuniversitario per le Biotecnologie, Trieste, Italy.
Abstract
PURPOSE: The aim of this study was to increase knowledge of genes associated with anorexia nervosa (AN) and their diagnostic offer, using a next generation sequencing (NGS) panel for the identification of genetic variants. The rationale underlying this test is that we first analyze the genes associated with syndromic forms of AN, then genes that were found to carry rare variants in AN patients who had undergone segregation analysis, and finally candidate genes intervening in the same molecular pathways or identified by GWAS or in mouse models. METHODS: We developed an NGS gene panel and used it to screen 68 Italian AN patients (63 females, 5 males). The panel included 162 genes. Family segregation study was conducted on available relatives of probands who reported significant genetic variants. RESULTS: In our analysis, we found potentially deleterious variants in 2 genes (PDE11A and SLC25A13) associated with syndromic forms of anorexia and predicted deleterious variants in the following 12 genes: CD36, CACNA1C, DRD4, EPHX2, ESR1, GRIN2A, GRIN3B, LRP2, NPY4R, PTGS2, PTPN22 and SGPP2. Furthermore, by Sanger sequencing of the promoter region of NNAT, we confirmed the involvement of this gene in the pathogenesis of AN. Family segregation studies further strengthened the possible causative role of CACNA1C, DRD4, GRIN2A, PTGS2, SGPP2, SLC25A13 and NNAT genes in AN etiology. CONCLUSION: The major finding of our study is the confirmation of the involvement of the NNAT gene in the pathogenesis of AN; furthermore, this study suggests that NGS-based testing can play an important role in the diagnostic evaluation of AN, excluding syndromic forms and increasing knowledge of the genetic etiology of AN. LEVEL OF EVIDENCE: Level I, experimental study.
PURPOSE: The aim of this study was to increase knowledge of genes associated with anorexia nervosa (AN) and their diagnostic offer, using a next generation sequencing (NGS) panel for the identification of genetic variants. The rationale underlying this test is that we first analyze the genes associated with syndromic forms of AN, then genes that were found to carry rare variants in AN patients who had undergone segregation analysis, and finally candidate genes intervening in the same molecular pathways or identified by GWAS or in mouse models. METHODS: We developed an NGS gene panel and used it to screen 68 Italian AN patients (63 females, 5 males). The panel included 162 genes. Family segregation study was conducted on available relatives of probands who reported significant genetic variants. RESULTS: In our analysis, we found potentially deleterious variants in 2 genes (PDE11A and SLC25A13) associated with syndromic forms of anorexia and predicted deleterious variants in the following 12 genes: CD36, CACNA1C, DRD4, EPHX2, ESR1, GRIN2A, GRIN3B, LRP2, NPY4R, PTGS2, PTPN22 and SGPP2. Furthermore, by Sanger sequencing of the promoter region of NNAT, we confirmed the involvement of this gene in the pathogenesis of AN. Family segregation studies further strengthened the possible causative role of CACNA1C, DRD4, GRIN2A, PTGS2, SGPP2, SLC25A13 and NNAT genes in AN etiology. CONCLUSION: The major finding of our study is the confirmation of the involvement of the NNAT gene in the pathogenesis of AN; furthermore, this study suggests that NGS-based testing can play an important role in the diagnostic evaluation of AN, excluding syndromic forms and increasing knowledge of the genetic etiology of AN. LEVEL OF EVIDENCE: Level I, experimental study.
Authors: V Precone; T Beccari; L Stuppia; M Baglivo; S Paolacci; E Manara; G A D Miggiano; B Falsini; A Trifirò; A Zanlari; K L Herbst; V Unfer; M Bertelli Journal: Eur Rev Med Pharmacol Sci Date: 2019-02 Impact factor: 3.507
Authors: M Rachele Ceccarini; Anna Tasegian; Marica Franzago; F Filomena Patria; Elisabetta Albi; Michela Codini; Carmela Conte; Matteo Bertelli; Laura Dalla Ragione; Liborio Stuppia; Tommaso Beccari Journal: Am J Med Genet B Neuropsychiatr Genet Date: 2019-11-20 Impact factor: 3.568
Authors: Hunna J Watson; Zeynep Yilmaz; Laura M Thornton; Christopher Hübel; Jonathan R I Coleman; Héléna A Gaspar; Julien Bryois; Anke Hinney; Virpi M Leppä; Manuel Mattheisen; Sarah E Medland; Stephan Ripke; Shuyang Yao; Paola Giusti-Rodríguez; Ken B Hanscombe; Kirstin L Purves; Roger A H Adan; Lars Alfredsson; Tetsuya Ando; Ole A Andreassen; Jessica H Baker; Wade H Berrettini; Ilka Boehm; Claudette Boni; Vesna Boraska Perica; Katharina Buehren; Roland Burghardt; Matteo Cassina; Sven Cichon; Maurizio Clementi; Roger D Cone; Philippe Courtet; Scott Crow; James J Crowley; Unna N Danner; Oliver S P Davis; Martina de Zwaan; George Dedoussis; Daniela Degortes; Janiece E DeSocio; Danielle M Dick; Dimitris Dikeos; Christian Dina; Monika Dmitrzak-Weglarz; Elisa Docampo; Laramie E Duncan; Karin Egberts; Stefan Ehrlich; Geòrgia Escaramís; Tõnu Esko; Xavier Estivill; Anne Farmer; Angela Favaro; Fernando Fernández-Aranda; Manfred M Fichter; Krista Fischer; Manuel Föcker; Lenka Foretova; Andreas J Forstner; Monica Forzan; Christopher S Franklin; Steven Gallinger; Ina Giegling; Johanna Giuranna; Fragiskos Gonidakis; Philip Gorwood; Monica Gratacos Mayora; Sébastien Guillaume; Yiran Guo; Hakon Hakonarson; Konstantinos Hatzikotoulas; Joanna Hauser; Johannes Hebebrand; Sietske G Helder; Stefan Herms; Beate Herpertz-Dahlmann; Wolfgang Herzog; Laura M Huckins; James I Hudson; Hartmut Imgart; Hidetoshi Inoko; Vladimir Janout; Susana Jiménez-Murcia; Antonio Julià; Gursharan Kalsi; Deborah Kaminská; Jaakko Kaprio; Leila Karhunen; Andreas Karwautz; Martien J H Kas; James L Kennedy; Anna Keski-Rahkonen; Kirsty Kiezebrink; Youl-Ri Kim; Lars Klareskog; Kelly L Klump; Gun Peggy S Knudsen; Maria C La Via; Stephanie Le Hellard; Robert D Levitan; Dong Li; Lisa Lilenfeld; Bochao Danae Lin; Jolanta Lissowska; Jurjen Luykx; Pierre J Magistretti; Mario Maj; Katrin Mannik; Sara Marsal; Christian R Marshall; Morten Mattingsdal; Sara McDevitt; Peter McGuffin; Andres Metspalu; Ingrid Meulenbelt; Nadia Micali; Karen Mitchell; Alessio Maria Monteleone; Palmiero Monteleone; Melissa A Munn-Chernoff; Benedetta Nacmias; Marie Navratilova; Ioanna Ntalla; Julie K O'Toole; Roel A Ophoff; Leonid Padyukov; Aarno Palotie; Jacques Pantel; Hana Papezova; Dalila Pinto; Raquel Rabionet; Anu Raevuori; Nicolas Ramoz; Ted Reichborn-Kjennerud; Valdo Ricca; Samuli Ripatti; Franziska Ritschel; Marion Roberts; Alessandro Rotondo; Dan Rujescu; Filip Rybakowski; Paolo Santonastaso; André Scherag; Stephen W Scherer; Ulrike Schmidt; Nicholas J Schork; Alexandra Schosser; Jochen Seitz; Lenka Slachtova; P Eline Slagboom; Margarita C T Slof-Op 't Landt; Agnieszka Slopien; Sandro Sorbi; Beata Świątkowska; Jin P Szatkiewicz; Ioanna Tachmazidou; Elena Tenconi; Alfonso Tortorella; Federica Tozzi; Janet Treasure; Artemis Tsitsika; Marta Tyszkiewicz-Nwafor; Konstantinos Tziouvas; Annemarie A van Elburg; Eric F van Furth; Gudrun Wagner; Esther Walton; Elisabeth Widen; Eleftheria Zeggini; Stephanie Zerwas; Stephan Zipfel; Andrew W Bergen; Joseph M Boden; Harry Brandt; Steven Crawford; Katherine A Halmi; L John Horwood; Craig Johnson; Allan S Kaplan; Walter H Kaye; James E Mitchell; Catherine M Olsen; John F Pearson; Nancy L Pedersen; Michael Strober; Thomas Werge; David C Whiteman; D Blake Woodside; Garret D Stuber; Scott Gordon; Jakob Grove; Anjali K Henders; Anders Juréus; Katherine M Kirk; Janne T Larsen; Richard Parker; Liselotte Petersen; Jennifer Jordan; Martin Kennedy; Grant W Montgomery; Tracey D Wade; Andreas Birgegård; Paul Lichtenstein; Claes Norring; Mikael Landén; Nicholas G Martin; Preben Bo Mortensen; Patrick F Sullivan; Gerome Breen; Cynthia M Bulik Journal: Nat Genet Date: 2019-07-15 Impact factor: 38.330
Authors: V Precone; S Paolacci; T Beccari; L Dalla Ragione; L Stuppia; M Baglivo; G Guerri; E Manara; G Tonini; K L Herbst; V Unfer; M Bertelli Journal: Eur Rev Med Pharmacol Sci Date: 2020-02 Impact factor: 3.507