Nonsense suppression induced readthrough of a novel PAX6 mutation in patient-derived cells of congenital aniridia

Mol Genet Genomic Med. 2020 May;8(5):e1198. doi: 10.1002/mgg3.1198. Epub 2020 Mar 3.

Abstract

Background: Congenital aniridia is a severe ocular abnormality characterized by incomplete formation of the iris and many other ocular complications. Most cases are caused by the paired box 6 (PAX6) gene mutations generating premature termination codons (PTCs).

Methods: Ophthalmic examination was performed on a Chinese pedigree with congenital aniridia. The mutation was identified by targeted next-generation sequencing. Nonsense suppression therapy was applied on patient-derived lymphocytes. The PAX6 expression was assayed by real-time polymerase chain reaction and Western blot.

Results: Complete aniridia was complicated with horizontal nystagmus, contract, foveal hypoplasia, and microphthalmia. A novel heterozygous c.702_703delinsAT (p.Tyr234*) mutation was found in exon 9 of PAX6, generating a PTC at the homeodomain. There were about 50% reductions of both full-length PAX6 protein and PAX6 mRNA in patient-derived lymphocytes, indicating haploinsufficiency due to nonsense-mediated mRNA decay. Ataluren (PTC124) and geneticin (G418) could induce about 30%-40% translational readthrough. Nonsense suppression therapy restored PAX6 protein to about 65%-70% of unaffected family controls.

Conclusion: Our data expanded the genetic and phenotypic variations of congenital aniridia, and showed the therapeutic effect of nonsense suppression on this disease using patient-derived cells.

Keywords: PAX6; congenital aniridia; nonsense suppression therapy; nonsense-mediated mRNA decay.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aniridia / genetics*
  • Aniridia / pathology
  • Cells, Cultured
  • Child
  • Female
  • Gentamicins / pharmacology
  • Haploinsufficiency
  • Heterozygote
  • Humans
  • INDEL Mutation*
  • Male
  • Middle Aged
  • Nonsense Mediated mRNA Decay / drug effects*
  • Oxadiazoles / pharmacology
  • PAX6 Transcription Factor / genetics*
  • PAX6 Transcription Factor / metabolism
  • Pedigree

Substances

  • Gentamicins
  • Oxadiazoles
  • PAX6 Transcription Factor
  • PAX6 protein, human
  • antibiotic G 418
  • ataluren

Associated data

  • GENBANK/NG_008679.1
  • GENBANK/NM_000280.4
  • GENBANK/NM_001101.5