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Arijit Mukhopadhyay, Konstantinos Nikopoulos, Alessandra Maugeri, Arjan P. M. de Brouwer, C. Eric van Nouhuys, Camiel J. F. Boon, Rahat Perveen, Hester A. A. Zegers, Dienke Wittebol-Post, Pieter R. van den Biesen, Saskia D. van der Velde-Visser, Han G. Brunner, Graeme C. M. Black, Carel B. Hoyng, Frans P. M. Cremers; Erosive Vitreoretinopathy and Wagner Disease Are Caused by Intronic Mutations in CSPG2/Versican That Result in an Imbalance of Splice Variants. Invest. Ophthalmol. Vis. Sci. 2006;47(8):3565-3572. doi: https://doi.org/10.1167/iovs.06-0141.
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purpose. Linkage intervals for erosive vitreoretinopathy (ERVR) and Wagner disease previously were found to overlap at 5q14.3. In a Japanese family with Wagner disease, a CSPG2/Versican splice site mutation (c.4004-2A→G) was recently reported that resulted in a 39-nucleotide exon 8 in-frame deletion. We investigated whether CSPG2/Versican was mutated in six Dutch families and one Chinese family with Wagner disease and in a family with ERVR.
methods. In all families, extensive ophthalmic examinations, haplotype analysis of the 5q14.3 region, and sequence analysis of CSPG2/Versican were performed. The effects of splice site mutations were assessed by reverse transcription–polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR (QPCR).
results. Three novel intron 7 sequence variants (c.4004-5T→C, c.4004-5T→A, c.4004-1G→A) were identified in seven families. The c.4004-5T→C variant was identified in four families with Wagner disease and a family with ERVR. The families were shown to carry the same 5q14.3 haplotype, strongly suggesting that this is a common Dutch founder variant. All three changes segregated with the disease in the respective families and were absent in 250 healthy individuals. In patients with the c.4004-5T→A and c.4004-1G→A variants, RT-PCR analysis of CSPG2/Versican showed activation of a cryptic splice site resulting in a 39-nt exon 8 in-frame deletion in splice variant V0. QPCR revealed a highly significant (P < 0.0001) and consistent increase of the V2 (>38-fold) and V3 (>12-fold) splice variants in all patients with intron 7 nucleotide changes and in a Chinese Wagner disease family, in which the genetic defect remains to be found.
conclusions. Wagner disease and ERVR are allelic disorders. Seven of the eight families exhibit a variant in intron 7 of CSPG2/Versican. The conspicuous clustering of sequence variants in the splice acceptor site of intron 7 and the consistent upregulation of the V2 and V3 isoforms strongly suggest that Wagner disease and ERVR may belong to a largely overlooked group of diseases that are caused by mRNA isoform balance shifts, representing a novel disease mechanism.
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