A targeted next-generation sequencing assay for the molecular diagnosis of genetic disorders with orodental involvement

Megana K. Prasad (1, 2) , Véronique Geoffroy (1, 2) , Serge Vicaire (3) , Bernard Jost (3) , Michael Dumas (3) , Stéphanie Le Gras (3) , Marzena Switala (4, 5) , Barbara Gasse (6) , Virginie Laugel-Haushalter (3) , Marie Paschaki (3, 1, 2) , Bruno Leheup (7) , Dominique Droz (7) , Amelie Dalstein (7) , Adeline Loing (4) , Bruno Grollemund (5) , Michèle Muller-Bolla (8, 9) , Serena Lopez-Cazaux (10) , Maryline Minoux (4, 5) , Sophie Jung (5, 4) , Frédéric Obry (5, 4) , Vincent Vogt (5, 4) , Jean-Luc Davideau (5) , Tiphaine Davit-Béal (6) , Anne-Sophie Kaiser (11) , Ute Moog (11) , Béatrice Richard (12, 13) , Jean-Jacques Morrier (12, 13) , Jean-Pierre Duprez (12, 13) , Sylvie Odent (14) , Isabelle Bailleul-Forestier (15, 16) , Monique Marie Rousset (17) , Laure Merametdijan (18, 19) , Annick Toutain (20) , Clara Joseph (21, 8) , Fabienne Giuliano (8) , Jean-Christophe Dahlet (4) , Aymeric Courval (22, 4) , Mustapha El Alloussi (23) , Samir Laouina (23) , Sylvie Soskin (4) , Nathalie Guffon (24) , Anne Dieux (17) , Bérénice Doray (25) , Stephanie Feierabend (26) , Emmanuelle Ginglinger (27) , Benjamin Fournier (28, 29, 29) , Muriel de La Dure Molla (28, 29, 29) , Yves Alembik (25) , Corinne Tardieu (30, 31) , François Clauss (5, 4) , Ariane Berdal (29, 28, 29) , Corinne Stoetzel (1, 2) , Marie Cécile Manière (5, 4) , Hélène Dollfus (32, 1, 2) , Agnès Bloch-Zupan (33, 3, 4)

1 LGM - Laboratoire de Génétique Médicale
2 FMTS - Fédération de Médecine Translationnelle de Strasbourg
3 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
4 HUS - Les Hôpitaux Universitaires de Strasbourg
5 UNISTRA - Université de Strasbourg
6 Evolution Paris Seine
7 CHRU Nancy - Centre Hospitalier Régional Universitaire de Nancy
8 CHU Nice - Centre Hospitalier Universitaire de Nice
9 URB2i (URP_4462) - Unité de Recherche Biomatériaux Innovants et Interfaces
10 Service d’Odontologie Conservatrice et Pédiatrique [CHU Nantes]
11 Universität Heidelberg [Heidelberg] = Heidelberg University
12 HCL - Hospices Civils de Lyon
13 UCBL - Université Claude Bernard Lyon 1
14 Service de génétique clinique [Rennes]
15 UT3 - Université Toulouse III - Paul Sabatier
16 CHU Toulouse - Centre Hospitalier Universitaire de Toulouse
17 CHRU Lille - Centre Hospitalier Régional Universitaire [CHU Lille]
18 CHU Nantes - Centre Hospitalier Universitaire de Nantes = Nantes University Hospital
19 Nantes Univ - Nantes Université
20 Service de génétique [Tours]
21 UNSA - Université de Nice Sophia-Antipolis
22 Nouvel Hôpital Civil de Strasbourg
23 UM5 - Université Mohammed V de Rabat [Agdal]
24 CHU Lyon
25 CHU Strasbourg - Centre Hospitalier Universitaire [Strasbourg]
26 Uniklinik - Universitäts Klinikum Freiburg = University Medical Center Freiburg
27 CH E.Muller Mulhouse - Centre Hospitalier Emile Muller [Mulhouse]
28 CRC (UMR_S_1138 / U1138) - Centre de Recherche des Cordeliers
29 CHU Rothschild [AP-HP]
30 ADES - Anthropologie bio-culturelle, Droit, Ethique et Santé
31 TIMONE - Hôpital de la Timone [CHU - APHM]
32 Centre de Référence pour les Affections Rares en Génétique Ophtalmologique (CARGO) et Service de Génétique Médicale
33 Faculté de chirurgie dentaire [Strasbourg]

Background Orodental diseases include several clinically and genetically heterogeneous disorders that can present in isolation or as part of a genetic syndrome. Due to the vast number of genes implicated in these disorders, establishing a molecular diagnosis can be challenging. We aimed to develop a targeted next-generation sequencing (NGS) assay to diagnose mutations and potentially identify novel genes mutated in this group of disorders. Methods We designed an NGS gene panel that targets 585 known and candidate genes in orodental disease. We screened a cohort of 101 unrelated patients without a molecular diagnosis referred to the Reference Centre for Oro-Dental Manifestations of Rare Diseases, Strasbourg, France, for a variety of orodental disorders including isolated and syndromic amelogenesis imperfecta (AI), isolated and syndromic selective tooth agenesis (STHAG), isolated and syndromic dentinogenesis imperfecta, isolated dentin dysplasia, otodental dysplasia and primary failure of tooth eruption. Results We discovered 21 novel pathogenic variants and identified the causative mutation in 39 unrelated patients in known genes (overall diagnostic rate: 39%). Among the largest subcohorts of patients with isolated AI (50 unrelated patients) and isolated STHAG (21 unrelated patients), we had a definitive diagnosis in 14 (27%) and 15 cases (71%), respectively. Surprisingly, COL17A1 mutations accounted for the majority of autosomal-dominant AI cases. Conclusions We have developed a novel targeted NGS assay for the efficient molecular diagnosis of a wide variety of orodental diseases. Furthermore, our panel will contribute to better understanding the contribution of these genes to orodental disease. Trial registration numbers NCT01746121 and NCT02397824

Mots clés: Genetic screening/counselling; Genetics; Genome-wide; Molecular genetics; Diagnostics tests

Domaines: Sciences du Vivant [q-bio]

Megana K. Prasad, Véronique Geoffroy, Serge Vicaire, Bernard Jost, Michael Dumas, et al.. A targeted next-generation sequencing assay for the molecular diagnosis of genetic disorders with orodental involvement. Journal of Medical Genetics, 2016, 53 (2), pp.98--110. ⟨10.1136/jmedgenet-2015-103302⟩. ⟨hal-01272929⟩