Molecular Diagnosis of Neonatal Diabetes Mellitus Using Next-Generation Sequencing of the Whole Exome
Open Access
- 25 October 2010
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 5 (10), e13630
- https://doi.org/10.1371/journal.pone.0013630
Abstract
Background: 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. Methodology/Principal Findings: 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. Conclusions/Significance: 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.Keywords
This publication has 14 references indexed in Scilit:
- A method and server for predicting damaging missense mutationsNature Methods, 2010
- Breakthroughs in monogenic diabetes genetics: from pediatric forms to young adulthood diabetes.2009
- Neonatal Diabetes MellitusEndocrine Reviews, 2008
- Heterozygous Missense Mutations in the Insulin Gene Are Linked to Permanent Diabetes Appearing in the Neonatal Period or in Early InfancyDiabetes, 2008
- Permanent Neonatal Diabetes Caused by Dominant, Recessive, or Compound Heterozygous SUR1 Mutations with Opposite Functional EffectsAmerican Journal of Human Genetics, 2007
- New ABCC8 Mutations in Relapsing Neonatal Diabetes and Clinical FeaturesDiabetes, 2007
- A genome-wide association study identifies novel risk loci for type 2 diabetesNature, 2007
- Switching from Insulin to Oral Sulfonylureas in Patients with Diabetes Due to Kir6.2 MutationsNew England Journal of Medicine, 2006
- Activating Mutations in theABCC8Gene in Neonatal Diabetes MellitusNew England Journal of Medicine, 2006
- Activating Mutations in the Gene Encoding the ATP-Sensitive Potassium-Channel Subunit Kir6.2 and Permanent Neonatal DiabetesNew England Journal of Medicine, 2004