title: ZNF408 is mutated in familial exudative vitreoretinopathy and is crucial for the development of zebrafish retinal vasculature.
abstract: Familial exudative vitreoretinopathy (FEVR) is a genetically heterogeneous disorder characterized by abnormal vascularization of the peripheral retina, which can result in retinal detachment and severe visual impairment. In a large Dutch FEVR family, we performed linkage analysis, exome sequencing, and segregation analysis of DNA variants. We identified putative disease-causing DNA variants in proline-alanine-rich ste20-related kinase (c.791dup; p.Ser265ValfsX64) and zinc finger protein 408 (ZNF408) (c.1363C>T; p.His455Tyr), the latter of which was also present in an additional Dutch FEVR family that subsequently appeared to share a common ancestor with the original family. Sequence analysis of ZNF408 in 132 additional individuals with FEVR revealed another potentially pathogenic missense variant, p.Ser126Asn, in a Japanese family. Immunolocalization studies in COS-1 cells transfected with constructs encoding the WT and mutant ZNF408 proteins, revealed that the WT and the p.Ser126Asn mutant protein show complete nuclear localization, whereas the p.His455Tyr mutant protein was localized almost exclusively in the cytoplasm. Moreover, in a cotransfection assay, the p.His455Tyr mutant protein retains the WT ZNF408 protein in the cytoplasm, suggesting that this mutation acts in a dominant-negative fashion. Finally, morpholino-induced knockdown of znf408 in zebrafish revealed defects in developing retinal and trunk vasculature, that could be rescued by coinjection of RNA encoding human WT ZNF408 but not p.His455Tyr mutant ZNF408. Together, our data strongly suggest that mutant ZNF408 results in abnormal retinal vasculogenesis in humans and is associated with FEVR.
authors.affiliation: Department of Human Genetics, Radboud University Medical Centre, 6500 HB Nijmegen, The Netherlands.
authors.forename: Rob W J
authors.initials: RW
authors.lastname: Collin
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Nikopoulos
authors.forename: Margo
authors.initials: M
authors.lastname: Dona
authors.forename: Christian
authors.initials: C
authors.lastname: Gilissen
authors.forename: Alexander
authors.initials: A
authors.lastname: Hoischen
authors.forename: F Nienke
authors.initials: FN
authors.lastname: Boonstra
authors.forename: James A
authors.initials: JA
authors.lastname: Poulter
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kondo
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Berger
authors.forename: Carmel
authors.initials: C
authors.lastname: Toomes
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tahira
authors.forename: Lucas R
authors.initials: LR
authors.lastname: Mohn
authors.forename: Ellen A
authors.initials: EA
authors.lastname: Blokland
authors.forename: Lisette
authors.initials: L
authors.lastname: Hetterschijt
authors.forename: Manir
authors.initials: M
authors.lastname: Ali
authors.forename: Johanne M
authors.initials: JM
authors.lastname: Groothuismink
authors.forename: Lonneke
authors.initials: L
authors.lastname: Duijkers
authors.forename: Chris F
authors.initials: CF
authors.lastname: Inglehearn
authors.forename: Lea
authors.initials: L
authors.lastname: Sollfrank
authors.forename: Tim M
authors.initials: TM
authors.lastname: Strom
authors.forename: Eiichi
authors.initials: E
authors.lastname: Uchio
authors.forename: C Erik
authors.initials: CE
authors.lastname: van Nouhuys
authors.forename: Hannie
authors.initials: H
authors.lastname: Kremer
authors.forename: Joris A
authors.initials: JA
authors.lastname: Veltman
authors.forename: Erwin
authors.initials: E
authors.lastname: van Wijk
authors.forename: Frans P M
authors.initials: FP
authors.lastname: Cremers
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005192:Family Health; D005260:Female; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D012171:Retinal Vessels / Q000196:embryology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D018630:Vitreoretinopathy, Proliferative / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D014157:Transcription Factors; C582974:ZNF408 protein, human; C582975:ZNF408 protein, zebrafish; D029961:Zebrafish Proteins