A <i>COL17A1</i> Splice-Altering Mutation Is Prevalent in Inherited Recurrent Corneal Erosions

Oliver, Verity F, van Bysterveldt, Katherine A, Cadzow, Murray, Steger, Bernhard, Romano, Vito ORCID: 0000-0002-5148-7643, Markie, David, Hewitt, Alex W, Mackey, David A, Willoughby, Colin E, Sherwin, Trevor
et al (show 3 more authors) (2016) A <i>COL17A1</i> Splice-Altering Mutation Is Prevalent in Inherited Recurrent Corneal Erosions. OPHTHALMOLOGY, 123 (4). pp. 709-722.

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<h4>Purpose</h4>Corneal dystrophies are a genetically heterogeneous group of disorders. We previously described a family with an autosomal dominant epithelial recurrent erosion dystrophy (ERED). We aimed to identify the underlying genetic cause of ERED in this family and 3 additional ERED families. We sought to characterize the potential function of the candidate genes using the human and zebrafish cornea.<h4>Design</h4>Case series study of 4 white families with a similar ERED. An experimental study was performed on human and zebrafish tissue to examine the putative biological function of candidate genes.<h4>Participants</h4>Four ERED families, including 28 affected and 17 unaffected individuals.<h4>Methods</h4>HumanLinkage-12 arrays (Illumina, San Diego, CA) were used to genotype 17 family members. Next-generation exome sequencing was performed on an uncle-niece pair. Segregation of potential causative mutations was confirmed using Sanger sequencing. Protein expression was determined using immunohistochemistry in human and zebrafish cornea. Gene expression in zebrafish was assessed using whole-mount in situ hybridization. Morpholino-induced transient gene knockdown was performed in zebrafish embryos.<h4>Main outcome measures</h4>Linkage microarray, exome analysis, DNA sequence analysis, immunohistochemistry, in situ hybridization, and morpholino-induced genetic knockdown results.<h4>Results</h4>Linkage microarray analysis identified a candidate region on chromosome chr10:12,576,562-112,763,135, and exploration of exome sequencing data identified 8 putative pathogenic variants in this linkage region. Two variants segregated in 06NZ-TRB1 with ERED: COL17A1 c.3156C→T and DNAJC9 c.334G→A. The COL17A1 c.3156C→T variant segregated in all 4 ERED families. We showed biologically relevant expression of these proteins in human cornea. Both proteins are expressed in the cornea of zebrafish embryos and adults. Zebrafish lacking Col17a1a and Dnajc9 during development show no gross corneal phenotype.<h4>Conclusions</h4>The COL17A1 c.3156C→T variant is the likely causative mutation in our recurrent corneal erosion families, and its presence in 4 independent families suggests that it is prevalent in ERED. This same COL17A1 c.3156C→T variant recently was identified in a separate pedigree with ERED. Our study expands the phenotypic spectrum of COL17A1 disease from autosomal recessive epidermolysis bullosa to autosomal dominant ERED and identifies COL17A1 as a key protein in maintaining integrity of the corneal epithelium.

Item Type: Article
Uncontrolled Keywords: Epithelium, Corneal, Animals, Zebrafish, Humans, Corneal Dystrophies, Hereditary, Disease Models, Animal, Non-Fibrillar Collagens, Autoantigens, Microscopy, Confocal, Immunohistochemistry, In Situ Hybridization, Reverse Transcriptase Polymerase Chain Reaction, Pedigree, Gene Expression Regulation, Gene Silencing, Alternative Splicing, Microsatellite Repeats, Mutation, Polymorphism, Single Nucleotide, Adolescent, Adult, Aged, Middle Aged, Child, Female, Male, HSP40 Heat-Shock Proteins, Genetic Linkage, High-Throughput Nucleotide Sequencing, Collagen Type XVII
Depositing User: Symplectic Admin
Date Deposited: 23 Feb 2017 08:17
Last Modified: 02 Jan 2024 08:26
DOI: 10.1016/j.ophtha.2015.12.008
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3006002