Journal
AMERICAN JOURNAL OF HUMAN GENETICS
Volume 102, Issue 4, Pages 517-527Publisher
CELL PRESS
DOI: 10.1016/j.ajhg.2018.02.008
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Funding
- FP7-PEOPLE-ITN programme EyeTN [317472]
- Macula Vision Research Foundation
- Rotterdamse Stichting Blindenbelangen
- Stichting Blindenhulp
- Stichting tot Verbetering van het Lot der Blinden
- Landelijke Stichting voor Blinden en Slechtzienden
- Macula Degeneratie fonds
- Stichting Blinden-Penning [Uitzicht 2016-12]
- Algemene Nederlandse Vereniging ter Voorkoming van Blindheid
- Stichting Blinden-Penning
- Stichting Oogfonds Nederland
- Stichting Macula Degeneratie Fonds
- Stichting Retina Nederland Fonds [UitZicht 2015-31]
- Stichting voor Ooglijders
- Stichting Dowilvo
- Stichting A.F. Deutman Researchfonds Oogheelkunde
- Foundation Fighting Blindness USA [PPA-0517-0717-RAD]
- National Eye Institute/NIH [EY019861, EY019007]
- Research to Prevent Blindness (New York, NY)
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Sequence analysis of the coding regions and splice site sequences in inherited retinal diseases is not able to uncover similar to 40% of the causal variants. Whole-genome sequencing can identify most of the non-coding variants, but their interpretation is still very challenging, in particular when the relevant gene is expressed in a tissue-specific manner. Deep-intronic variants in ABCA4 have been associated with autosomal-recessive Stargardt disease (STGD1), but the exact pathogenic mechanism is unknown. By generating photoreceptor precursor cells (PPCs) from fibroblasts obtained from individuals with STGD1, we demonstrated that two neighboring deep-intronic ABCA4 variants (c.4539thorn2001G>A and c.4539thorn2028C>T) result in a retina-specific 345-nt pseudoexon insertion (predicted protein change: p.Arg1514Leufs*36), likely due to the creation of exonic enhancers. Administration of antisense oligonucleotides (AONs) targeting the 345-nt pseudoexon can significantly rescue the splicing defect observed in PPCs of two individuals with these mutations. Intriguingly, an AON that is complementary to c.4539thorn2001G>A rescued the splicing defect only in PPCs derived from an individual with STGD1 with this but not the other mutation, demonstrating the high specificity of AONs. In addition, a single AON molecule rescued splicing defects associated with different neighboring mutations, thereby providing new strategies for the treatment of persons with STGD1. As many genes associated with human genetic conditions are expressed in specific tissues and pre-mRNA splicing may also rely on organ-specific factors, our approach to investigate and treat splicing variants using differentiated cells derived from individuals with STGD1 can be applied to any tissue of interest.
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