The utility of massively parallel sequencing for posterior polymorphous corneal dystrophy type 3 molecular diagnosis
Language English Country Great Britain, England Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
30851240
DOI
10.1016/j.exer.2019.03.002
PII: S0014-4835(18)30789-9
Knihovny.cz E-resources
- Keywords
- Aberrant splicing, Breakpoint mapping, Exome, Genome, Massively parallel sequencing, Posterior polymorphous corneal dystrophy type 3, ZEB1,
- MeSH
- Corneal Dystrophies, Hereditary diagnosis genetics metabolism MeSH
- Child MeSH
- DNA genetics MeSH
- Adult MeSH
- Exons MeSH
- Heterozygote MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Mutation * MeSH
- DNA Mutational Analysis MeSH
- Child, Preschool MeSH
- Pedigree MeSH
- Base Sequence MeSH
- Sequence Deletion MeSH
- Aged MeSH
- Zinc Finger E-box-Binding Homeobox 1 genetics metabolism MeSH
- High-Throughput Nucleotide Sequencing MeSH
- Zinc Fingers MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Child, Preschool MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA MeSH
- Zinc Finger E-box-Binding Homeobox 1 MeSH
- ZEB1 protein, human MeSH Browser
The aim of this study was to identify the molecular genetic cause of disease in posterior polymorphous corneal dystrophy (PPCD) probands of diverse origin and to assess the utility of massively parallel sequencing in the detection of ZEB1 mutations. We investigated a total of 12 families (five British, four Czech, one Slovak and two Swiss). Ten novel and two recurrent disease-causing mutations in ZEB1, were identified in probands by Sanger (n = 5), exome (n = 4) and genome (n = 3) sequencing. Sanger sequencing was used to confirm the mutations detected by massively parallel sequencing, and to perform segregation analysis. Genome sequencing revealed that one proband harboured a novel ∼0.34 Mb heterozygous de novo deletion spanning exons 1-7 and part of exon 8. Transcript analysis confirmed that the ZEB1 transcript is detectable in blood-derived RNA samples and that the disease-associated variant c.482-2A>G leads to aberrant pre-mRNA splicing. De novo mutations, which are a feature of PPCD3, were found in the current study with an incidence rate of at least 16.6%. In general, massively parallel sequencing is a time-efficient way to detect PPCD3-associated mutations and, importantly, genome sequencing enables the identification of full or partial heterozygous ZEB1 deletions that can evade detection by both Sanger and exome sequencing. These findings contribute to our understanding of PPCD3, for which currently, 49 pathogenic variants have been identified, all of which are predicted to be null alleles.
Center for Eye Microsurgery Gagarinova 7 B 821 03 Bratislava Slovakia
Moorfields Eye Hospital 162 City Road EC1V 2PD London United Kingdom
Ophthalmology Clinic of Dr Tomas Kubena U Zimniho Stadionu 1759 760 00 Zlin Czech Republic
UCL Institute of Ophthalmology 11 43 Bath Street EC1V 9EL London United Kingdom
References provided by Crossref.org
CUGC for posterior polymorphous corneal dystrophy (PPCD)
