Exome sequencing coupled with mRNA analysis identifies NDUFAF6 as a Leigh gene

Mol Genet Metab. 2016 Nov;119(3):214-222. doi: 10.1016/j.ymgme.2016.09.001. Epub 2016 Sep 3.

Abstract

We report here the case of a young male who started to show verbal fluency disturbance, clumsiness and gait anomalies at the age of 3.5years and presented bilateral striatal necrosis. Clinically, the diagnosis was compatible with Leigh syndrome but the underlying molecular defect remained elusive even after exome analysis using autosomal/X-linked recessive or de novo models. Dosage of respiratory chain activity on fibroblasts, but not in muscle, underlined a deficit in complex I. Re-analysis of heterozygous probably pathogenic variants, inherited from one healthy parent, identified the p.Ala178Pro in NDUFAF6, a complex I assembly factor. RNA analysis showed an almost mono-allelic expression of the mutated allele in blood and fibroblasts and puromycin treatment on cultured fibroblasts did not lead to the rescue of the maternal allele expression, not supporting the involvement of nonsense-mediated RNA decay mechanism. Complementation assay underlined a recovery of complex I activity after transduction of the wild-type gene. Since the second mutation was not detected and promoter methylation analysis resulted normal, we hypothesized a non-exonic event in the maternal allele affecting a regulatory element that, in conjunction with the paternal mutation, leads to the autosomal recessive disorder and the different allele expression in various tissues. This paper confirms NDUFAF6 as a genuine morbid gene and proposes the coupling of exome sequencing with mRNA analysis as a method useful for enhancing the exome sequencing detection rate when the simple application of classical inheritance models fails.

Keywords: Exome sequencing; Leigh syndrome; NDUFAF6; RNA analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Child, Preschool
  • Exome / genetics*
  • Heterozygote
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Leigh Disease / genetics*
  • Leigh Disease / physiopathology
  • Male
  • Mitochondrial Proteins / genetics*
  • Mutation
  • Pedigree
  • Phenotype
  • RNA, Messenger / genetics
  • Speech Disorders / genetics*
  • Speech Disorders / physiopathology
  • Striatonigral Degeneration / congenital
  • Striatonigral Degeneration / genetics
  • Striatonigral Degeneration / physiopathology

Substances

  • Mitochondrial Proteins
  • NDUFAF6 protein, human
  • RNA, Messenger

Supplementary concepts

  • Striatonigral degeneration infantile