Next-generation sequencing and novel variant determination in a cohort of 92 familial exudative vitreoretinopathy patients

Invest Ophthalmol Vis Sci. 2015 Feb 24;56(3):1937-46. doi: 10.1167/iovs.14-16065.

Abstract

Purpose: Familial exudative vitreoretinopathy (FEVR) is a developmental disease that can cause visual impairment and retinal detachment at a young age. Four genes involved in the Wnt signaling pathway were previously linked to this disease: NDP, FDZ4, LRP5, and TSPAN12. Identification of novel disease-causing alleles allows for a deeper understanding of the disease, better molecular diagnosis, and improved treatment.

Methods: Sequencing libraries from 92 FEVR patients were generated using a custom capture panel to enrich for 163 known retinal disease-causing genes in humans. Samples were processed using next generation sequencing (NGS) techniques followed by data analysis to identify and classify single nucleotide variants and small insertions and deletions. Sanger validation and segregation testing were used to verify suspected variants.

Results: Of the cohort of 92, 45 patients were potentially solved (48.9%). Solved cases resulted from the determination of 49 unique mutations, 41 of which are novel. Of the novel variants discovered, 13 were highly likely to cause FEVR due to the nature of these variants (frameshifting indels, splicing mutations, and nonsense variants types). To our knowledge, this is the largest study of a FEVR cohort using NGS.

Conclusions: We were able to determine probable disease-causing variants in a large number of FEVR patients, the majority of which were novel. Knowledge of these variants will help to further characterize and diagnose FEVR.

Keywords: FEVR; familial segregation; next-generation sequencing; novel alleles.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cohort Studies
  • DNA Mutational Analysis / methods*
  • DNA-Binding Proteins / genetics
  • Eye Diseases, Hereditary
  • Eye Proteins / genetics
  • Familial Exudative Vitreoretinopathies
  • Female
  • Frizzled Receptors / genetics
  • Genetic Predisposition to Disease
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Low Density Lipoprotein Receptor-Related Protein-5 / genetics
  • Male
  • Mutation*
  • Nerve Tissue Proteins / genetics
  • Retinal Diseases / genetics
  • Sequence Analysis, DNA / methods*
  • Tetraspanins / genetics
  • Transcription Factors / genetics

Substances

  • DNA-Binding Proteins
  • Eye Proteins
  • FZD4 protein, human
  • Frizzled Receptors
  • LRP5 protein, human
  • Low Density Lipoprotein Receptor-Related Protein-5
  • NDP protein, human
  • Nerve Tissue Proteins
  • Tetraspanins
  • Transcription Factors
  • ZNF408 protein, human