title: Deep Intronic Sequence Variants in COL2A1 Affect the Alternative Splicing Efficiency of Exon 2, and May Confer a Risk for Rhegmatogenous Retinal Detachment.
abstract: COL2A1 mutations causing haploinsufficiency of type II collagen cause type 1 Stickler syndrome that has a high risk of retinal detachment and failure of the vitreous to develop normally. Exon 2 of COL2A1 is alternatively spliced, expressed in the eye but not in mature cartilage and encodes a region that binds growth factors TGFβ1 and BMP-2. We investigated how both an apparently de novo variant and a polymorphism in intron 2 altered the efficiency of COL2A1 exon 2 splicing and how the latter may act as a predisposing risk factor for the occurrence of posterior vitreous detachment (PVD)-associated rhegmatogenous retinal detachment (RRD) in the general population. Using amplification of illegitimate transcripts and allele-specific minigenes expressed in cultured cells, we demonstrate variability in exon 2 inclusion not only between different control individuals, but also between different COL2A1 alleles. We identify transacting factors that bind to allele-specific RNA sequences, and investigate the effect of knockdown and overexpression of these factors on exon 2 splicing efficiency. Finally, using a specific cohort of patients with PVD-associated RRD and a control population, we demonstrate a significant difference in the frequency of the COL2A1 intronic variant rs1635532 between the two groups.
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.|Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK.
authors.forename: Carl
authors.initials: C
authors.lastname: Spickett
authors.affiliation: Academic Unit Ophthalmology, King's College London, London, SE1 7EH, UK.
authors.forename: Pirro
authors.initials: P
authors.lastname: Hysi
authors.affiliation: Academic Unit Ophthalmology, King's College London, London, SE1 7EH, UK.
authors.forename: Chistopher J
authors.initials: CJ
authors.lastname: Hammond
authors.affiliation: College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Prescott
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Fincham
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.
authors.forename: Arabella V
authors.initials: AV
authors.lastname: Poulson
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.|Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK.|Regional Molecular Genetics Laboratory, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.
authors.forename: Annie M
authors.initials: AM
authors.lastname: McNinch
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK. ar204@cam.ac.uk.|Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK. ar204@cam.ac.uk.|Regional Molecular Genetics Laboratory, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK. ar204@cam.ac.uk.
authors.forename: Allan J
authors.initials: AJ
authors.lastname: Richards
authors.affiliation: Vitreoretinal Research Group, Cambridge University NHS Foundation Trust, Cambridge, CB2 0QQ, UK.
authors.forename: Martin P
authors.initials: MP
authors.lastname: Snead
mesh_terms: D017398:Alternative Splicing; D002478:Cells, Cultured; D024043:Collagen Type II / Q000235:genetics*; D005091:Exons; D015785:Eye Diseases, Hereditary / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D006801:Humans; D007438:Introns; D009154:Mutation; D012163:Retinal Detachment / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: C499177:COL2A1 protein, human; D024043:Collagen Type II
keywords: COL2A1; alternative splicing; giant retinal tear; posterior vitreous detachment; retinal detachment