Journal Article

Molecular Diagnosis of Neonatal Diabetes Mellitus Using Next-Generation Sequencing of the Whole Exome

Létourneau, Louis, Sladek, Robert, Scharfmann, Raphael, Busiah, Kanetee, Vaxillaire, Martine, De Graeve, Franck, Sand, Olivier, Bonnefond, Amélie, Polak, Michel, Bellanné-Chantelot, Christine, Gallina, Sophie, Delplanque, Jérôme, Simon, Albane, Durand, Emmanuelle, Lobbens, Stéphane, Froguel, Philippe

2010

Overview

Accurate molecular diagnosis of monogenic non-autoimmune neonatal diabetes mellitus (NDM) is critical for patient care, as patients carrying a mutation in KCNJ11 or ABCC8 can be treated by oral sulfonylurea drugs instead of insulin therapy. This diagnosis is currently based on Sanger sequencing of at least 42 PCR fragments from the KCNJ11, ABCC8, and INS genes. Here, we assessed the feasibility of using the next-generation whole exome sequencing (WES) for the NDM molecular diagnosis. We carried out WES for a patient presenting with permanent NDM, for whom mutations in KCNJ11, ABCC8 and INS and abnormalities in chromosome 6q24 had been previously excluded. A solution hybridization selection was performed to generate WES in 76 bp paired-end reads, by using two channels of the sequencing instrument. WES quality was assessed using a high-resolution oligonucleotide whole-genome genotyping array. From our WES with high-quality reads, we identified a novel non-synonymous mutation in ABCC8 (c.1455G>C/p.Q485H), despite a previous negative sequencing of this gene. This mutation, confirmed by Sanger sequencing, was not present in 348 controls and in the patient's mother, father and young brother, all of whom are normoglycemic. WES identified a novel de novo ABCC8 mutation in a NDM patient. Compared to the current Sanger protocol, WES is a comprehensive, cost-efficient and rapid method to identify mutations in NDM patients. We suggest WES as a near future tool of choice for further molecular diagnosis of NDM cases, negative for chr6q24, KCNJ11 and INS abnormalities.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Abnormalities / Chromosome 6 / Chromosomes, Human, Pair 6 / Deoxyribonucleic acid / Diabetes / Diabetes and Endocrinology / Diabetes and Endocrinology/Type 2 Diabetes / Diabetes mellitus / Diabetes Mellitus - diagnosis / Diabetes Mellitus - genetics / Diabetes therapy / Diagnosis / DNA / DNA sequencing / Drugs / Endocrinology / Endocrinology and metabolism / Exons / Family medical history / Feasibility studies / Gene loci / Gene sequencing / Genes / Genetics / Genetics and Genomics / Genetics and Genomics/Gene Discovery / Genetics and Genomics/Genetics of Disease / Genetics and Genomics/Medical Genetics / Genomes / Genomics / Genotyping / Human genetics / Human health and pathology / Humans / Hybridization / Infant, Newborn / Infant, Newborn, Diseases - diagnosis / Infant, Newborn, Diseases - genetics / Insulin / Life Sciences / Medical diagnosis / Molecular Diagnostic Techniques / Mutation / Neonates / Newborn babies / Newborn infants / Oligonucleotides / Patients / Pediatrics / Polymorphism, Single Nucleotide / Potassium / Quality assessment / Sulfonylurea