Suárez-Herrera, Nuria;
Riswick, Iris B;
Vázquez-Domínguez, Irene;
Duijkers, Lonneke;
Karjosukarso, Dyah W;
Piccolo, Davide;
Bauwens, Miriam;
... Collin, Rob WJ; + view all
(2024)
Proof-of-concept for multiple AON delivery by a single U7snRNA vector to restore splicing defects in ABCA4.
Molecular Therapy
, 32
(3)
pp. 837-851.
10.1016/j.ymthe.2024.01.019.
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1-s2.0-S1525001624000194-main.pdf - Accepted Version Access restricted to UCL open access staff until 7 March 2025. Download (7MB) |
Abstract
The high allelic heterogeneity in Stargardt disease (STGD1) complicates the design of intervention strategies. A significant proportion of pathogenic intronic ABCA4 variants alters the pre-mRNA splicing process. Antisense oligonucleotides (AONs) are an attractive yet mutation-specific therapeutic strategy to restore these splicing defects. In this study, we experimentally assessed the potential of a splicing modulation therapy to target multiple intronic ABCA4 variants. AONs were inserted into U7snRNA gene cassettes and tested in midigene-based splice assays. Five potent antisense sequences were selected to generate a multiple U7snRNA cassette construct, and this combination vector showed substantial rescue of all of the splicing defects. Therefore, the combination cassette was used for viral synthesis and assessment in patient-derived photoreceptor precursor cells (PPCs). Simultaneous delivery of several modified U7snRNAs through a single AAV, however, did not show substantial splicing correction, probably due to suboptimal transduction efficiency in PPCs and/or a heterogeneous viral population containing incomplete AAV genomes. Overall, these data demonstrate the potential of the U7snRNA system to rescue multiple splicing defects, but also suggest that AAV-associated challenges are still a limiting step, underscoring the need for further optimization before implementing this strategy as a potential treatment for STGD1.
| Type: | Article |
|---|---|
| Title: | Proof-of-concept for multiple AON delivery by a single U7snRNA vector to restore splicing defects in ABCA4 |
| Location: | United States |
| DOI: | 10.1016/j.ymthe.2024.01.019 |
| Publisher version: | https://doi.org/10.1016/j.ymthe.2024.01.019 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher's terms and conditions. |
| Keywords: | U7snRNA, antisense oligonucleotide, splicing modulation, AAV, RNA therapy, ABCA4, Stargardt disease, inherited retinal disease, patient-derived photoreceptor precursor cells |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10186131 |
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