Bénédicte Sudrié-Arnaud1, Florent Marguet2, Sophie Patrier3, Jelena Martinovic4, Ferielle Louillet5, Françoise Broux5, Françoise Charbonnier6, Hélène Dranguet1, Sophie Coutant6, Myriam Vezain6, Raphaël Lanos6, Abdellah Tebani1, Maria Fuller7, Foudil Lamari8, Pascal Chambon9, Anne-Claire Brehin9, Laetitia Trestard10, Isabelle Tournier6, Stéphane Marret11, Eric Verspyck12, Annie Laquerrière2, Soumeya Bekri13. 1. Department of Metabolic Biochemistry, Rouen University Hospital, Rouen 76000, France. 2. Department of Pathology, Rouen University Hospital, Rouen 76000, France; Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France. 3. Department of Pathology, Rouen University Hospital, Rouen 76000, France. 4. Unit of Fetal Pathology, Antoine Beclere Hospital, South Paris University, Clamart, France. 5. Department of Pediatrics, Rouen University Hospital, 76000 Rouen, France. 6. Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France. 7. Genetics and Molecular Pathology, SA Pathology [at Women's and Children's Hospital], 72 King William Road, North Adelaide, South Australia 5006, Australia; Department of Pediatrics, University of Adelaide, Adelaide, South Australia 5005, Australia. 8. Bioclinic and Genetic Unit of Neurometabolic Diseases, Pitié-Salpêtrière Hospital, APHP, Paris 75013, France. 9. Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France; Department of Genetics, Normandy Centre for Genomic and Personalized Medicine, Rouen University Hospital, Rouen 76000, France. 10. Department of Obstetrics and Gynecology, Belvedere Hospital, Mont-Saint-Aignan, France. 11. Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France; Department of Neonatal Pediatrics, Intensive Care and Neuropediatrics, Rouen University Hospital, 76000 Rouen, France. 12. Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France; Department of Obstetrics and Gynecology, Rouen University Hospital, 76000 Rouen, France. 13. Department of Metabolic Biochemistry, Rouen University Hospital, Rouen 76000, France; Normandie Univ, UNIROUEN, CHU Rouen, INSERM U1245, 76000 Rouen, France. Electronic address: soumeya.bekri@chu-rouen.fr.
Abstract
PURPOSES: Hydrops fetalis is a life-threatening fetal condition, and 85% of all cases are classified as nonimmune hydrops fetalis (NIHF). Up to 15% of NIHF cases may be due to inborn errors of metabolism (IEM), but a large proportion of cases linked to metabolic disorders remains undiagnosed. This lack of diagnosis may be related to the limitations of conventional biological procedures, which involve sequential investigations and require multiple samples and steps. In addition, this approach is time consuming. We have developed a next-generation sequencing (NGS) panel to investigate metabolic causes of NIHF, ascites, and polyhydramnios associated to another fetal abnormality. METHODS: The hydrops fetalis (HydFet) panel was designed to cover the coding regions and flanking intronic sequences of 41 genes. A retrospective study of amniotic fluid samples from 40 subjects was conducted. A prospective study was subsequently initiated, and six samples were analyzed using the NGS panel. RESULTS: Five IEM diagnoses were made using the HydFet panel (Niemann-Pick type C (NPC), Barth syndrome, HNF1Β deficiency, GM1 gangliosidosis, and Gaucher disease). This analysis also allowed the identification of 8p sequence triplication in an additional case. CONCLUSION: NGS combined with robust bioinformatics analyses is a useful tool for identifying the causative variants of NIHF. Subsequent functional characterization of the protein encoded by the altered gene and morphological studies may confirm the diagnosis. This paradigm shift allows a significant improvement of IEM diagnosis in NIHF.
PURPOSES: Hydrops fetalis is a life-threatening fetal condition, and 85% of all cases are classified as nonimmune hydrops fetalis (NIHF). Up to 15% of NIHF cases may be due to inborn errors of metabolism (IEM), but a large proportion of cases linked to metabolic disorders remains undiagnosed. This lack of diagnosis may be related to the limitations of conventional biological procedures, which involve sequential investigations and require multiple samples and steps. In addition, this approach is time consuming. We have developed a next-generation sequencing (NGS) panel to investigate metabolic causes of NIHF, ascites, and polyhydramnios associated to another fetal abnormality. METHODS: The hydrops fetalis (HydFet) panel was designed to cover the coding regions and flanking intronic sequences of 41 genes. A retrospective study of amniotic fluid samples from 40 subjects was conducted. A prospective study was subsequently initiated, and six samples were analyzed using the NGS panel. RESULTS: Five IEM diagnoses were made using the HydFet panel (Niemann-Pick type C (NPC), Barth syndrome, HNF1Β deficiency, GM1 gangliosidosis, and Gaucher disease). This analysis also allowed the identification of 8p sequence triplication in an additional case. CONCLUSION: NGS combined with robust bioinformatics analyses is a useful tool for identifying the causative variants of NIHF. Subsequent functional characterization of the protein encoded by the altered gene and morphological studies may confirm the diagnosis. This paradigm shift allows a significant improvement of IEM diagnosis in NIHF.
Authors: Huda B Al-Kouatly; Laura Felder; Mona M Makhamreh; Stephanie L Kass; Neeta L Vora; Vincenzo Berghella; Seth Berger; David A Wenger; Paola Luzi Journal: Prenat Diagn Date: 2020-03-20 Impact factor: 3.050