Exome sequencing for simultaneous mutation screening in children with hemophagocytic lymphohistiocytosis

Int J Hematol. 2017 Aug;106(2):282-290. doi: 10.1007/s12185-017-2223-3. Epub 2017 Mar 28.

Abstract

In the present study, we used exome sequencing to analyze PRF1, UNC13D, STX11, and STXBP2, as well as genes associated with primary immunodeficiency disease (RAB27A, LYST, AP3B1, SH2D1A, ITK, CD27, XIAP, and MAGT1) in Thai children with hemophagocytic lymphohistiocytosis (HLH). We performed mutation analysis of HLH-associated genes in 25 Thai children using an exome sequencing method. Genetic variations found within these target genes were compared to exome sequencing data from 133 healthy individuals. Variants identified with minor allele frequencies <5% and novel mutations were confirmed using Sanger sequencing. Exome sequencing data revealed 101 non-synonymous single nucleotide polymorphisms (SNPs) in all subjects. These SNPs were classified as pathogenic (n = 1), likely pathogenic (n = 16), variant of unknown significance (n = 12), or benign variant (n = 72). Homozygous, compound heterozygous, and double-gene heterozygous variants, involving mutations in PRF1 (n = 3), UNC13D (n = 2), STXBP2 (n = 3), LYST (n = 3), XIAP (n = 2), AP3B1 (n = 1), RAB27A (n = 1), and MAGT1 (n = 1), were demonstrated in 12 patients. Novel mutations were found in most patients in this study. In conclusion, exome sequencing demonstrated the ability to identify rare genetic variants in HLH patients. This method is useful in the detection of mutations in multi-gene associated diseases.

Keywords: Exome sequencing; Hemophagocytic lymphohistiocytosis; Molecular diagnosis; Mutation screening; Next-generation sequencing.

MeSH terms

  • Child
  • DNA Mutational Analysis / methods*
  • Exome / genetics*
  • Genetic Association Studies*
  • Genetic Testing / methods*
  • Humans
  • Lymphohistiocytosis, Hemophagocytic / diagnosis*
  • Lymphohistiocytosis, Hemophagocytic / genetics
  • Membrane Proteins / genetics
  • Molecular Diagnostic Techniques
  • Multigene Family / genetics
  • Mutation*
  • Perforin / genetics
  • Polymorphism, Single Nucleotide
  • Qa-SNARE Proteins / genetics
  • Sequence Analysis, DNA / methods*
  • Vesicular Transport Proteins / genetics
  • rab GTP-Binding Proteins / genetics
  • rab27 GTP-Binding Proteins

Substances

  • LYST protein, human
  • Membrane Proteins
  • PRF1 protein, human
  • Qa-SNARE Proteins
  • STX11 protein, human
  • UNC13D protein, human
  • Vesicular Transport Proteins
  • rab27 GTP-Binding Proteins
  • Perforin
  • RAB27A protein, human
  • rab GTP-Binding Proteins