Identification and functional analysis of novel FZD4 mutations in Han Chinese with familial exudative vitreoretinopathy

Sci Rep. 2015 Nov 4:5:16120. doi: 10.1038/srep16120.

Abstract

Familial exudative vitreoretinopathy (FEVR) is a hereditary eye disease characterized by defects in the development of retinal vessels. However, known genetic mutations can only explain approximately 50% of FEVR patients. To assess the mutation frequency of Frizzled 4 (FZD4) in Chinese patients, we analysed patients with FEVR from 61 families from China to identify mutations in FZD4 and to study the effects of identified mutations on FZD4 function. All coding exons and adjacent intronic regions of FZD4 were amplified by polymerase chain reaction and subjected to Sanger sequencing analysis. Three mutations in the FZD4 gene were identified in these families. Of these, two were novel mutations: p.E134* and p.T503fs. Both mutations involve highly conserved residues and were not present in 800 normal individuals. Each of these two novel FZD4 mutations was introduced into wild-type FZD4 cDNA by site-directed mutagenesis. Wild-type and mutant FZD4 DNAs were introduced into HEK293 cells to analyse the function of FZD4 in Norrin-dependent activation of the Norrin/β-catenin pathway using luciferase reporter assays. Both the p.E134* and p.T503fs mutants failed to induce luciferase reporter activity in response to Norrin. Our study identified two novel FZD4 mutations in Chinese patients with FEVR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Angiography
  • Asian People / genetics*
  • Base Sequence
  • Child, Preschool
  • China
  • Eye Diseases, Hereditary
  • Eye Proteins / metabolism
  • Familial Exudative Vitreoretinopathies
  • Female
  • Frizzled Receptors / genetics*
  • Frizzled Receptors / metabolism
  • Genotype
  • HEK293 Cells
  • Humans
  • Male
  • Mutagenesis, Site-Directed
  • Nerve Tissue Proteins / metabolism
  • Pedigree
  • Phenotype
  • Polymorphism, Single Nucleotide
  • Retinal Diseases / genetics*
  • Retinal Diseases / pathology
  • Wnt Signaling Pathway
  • beta Catenin / metabolism

Substances

  • Eye Proteins
  • FZD4 protein, human
  • Frizzled Receptors
  • NDP protein, human
  • Nerve Tissue Proteins
  • beta Catenin