Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells
Leber congenital amaurosis is a severe hereditary retinal dystrophy responsible for neonatal blindness. The most common disease-causing mutation (c.2991+1655A>G; 10–15%) creates a strong splice donor site that leads to insertion of a cryptic exon encoding a premature stop codon. Recently, we repo...
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S216225311630035X |
| _version_ | 1818509813480423424 |
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| author | Xavier Gérard Isabelle Perrault Arnold Munnich Josseline Kaplan Jean-Michel Rozet |
| author_facet | Xavier Gérard Isabelle Perrault Arnold Munnich Josseline Kaplan Jean-Michel Rozet |
| author_sort | Xavier Gérard |
| collection | DOAJ |
| description | Leber congenital amaurosis is a severe hereditary retinal dystrophy responsible for neonatal blindness. The most common disease-causing mutation (c.2991+1655A>G; 10–15%) creates a strong splice donor site that leads to insertion of a cryptic exon encoding a premature stop codon. Recently, we reported that splice-switching oligonucleotides (SSO) allow skipping of the mutant cryptic exon and the restoration of ciliation in fibroblasts of affected patients, supporting the feasibility of a SSO-mediated exon skipping strategy to correct the aberrant splicing. Here, we present data in the wild-type mouse, which demonstrate that intravitreal administration of 2’-OMePS-SSO allows selective alteration of Cep290 splicing in retinal cells, including photoreceptors as shown by successful alteration of Abca4 splicing using the same approach. We show that both SSOs and Cep290 skipped mRNA were detectable for at least 1 month and that intravitreal administration of oligonucleotides did not provoke any serious adverse event. These data suggest that intravitreal injections of SSO should be considered to bypass protein truncation resulting from the c.2991+1655A>G mutation as well as other truncating mutations in genes which like CEP290 or ABCA4 have a mRNA size that exceed cargo capacities of US Food and Drug Administration (FDA)-approved adeno-associated virus (AAV)-vectors, thus hampering gene augmentation therapy. |
| first_indexed | 2024-12-10T22:50:34Z |
| format | Article |
| id | doaj.art-779dc79240cf4246a7e9476f563372ff |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-10T22:50:34Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-779dc79240cf4246a7e9476f563372ff2022-12-22T01:30:26ZengElsevierMolecular Therapy: Nucleic Acids2162-25312015-01-014C10.1038/mtna.2015.24Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal CellsXavier Gérard0Isabelle Perrault1Arnold Munnich2Josseline Kaplan3Jean-Michel Rozet4Laboratory of Genetics in Ophthalmology, Inserm UMR1163, Institut Imagine, Université Paris Descartes Sorbonne Paris Cité, Hôpital Necker, Paris, FranceLaboratory of Genetics in Ophthalmology, Inserm UMR1163, Institut Imagine, Université Paris Descartes Sorbonne Paris Cité, Hôpital Necker, Paris, FranceLaboratory of Genetics in Ophthalmology, Inserm UMR1163, Institut Imagine, Université Paris Descartes Sorbonne Paris Cité, Hôpital Necker, Paris, FranceLaboratory of Genetics in Ophthalmology, Inserm UMR1163, Institut Imagine, Université Paris Descartes Sorbonne Paris Cité, Hôpital Necker, Paris, FranceLaboratory of Genetics in Ophthalmology, Inserm UMR1163, Institut Imagine, Université Paris Descartes Sorbonne Paris Cité, Hôpital Necker, Paris, FranceLeber congenital amaurosis is a severe hereditary retinal dystrophy responsible for neonatal blindness. The most common disease-causing mutation (c.2991+1655A>G; 10–15%) creates a strong splice donor site that leads to insertion of a cryptic exon encoding a premature stop codon. Recently, we reported that splice-switching oligonucleotides (SSO) allow skipping of the mutant cryptic exon and the restoration of ciliation in fibroblasts of affected patients, supporting the feasibility of a SSO-mediated exon skipping strategy to correct the aberrant splicing. Here, we present data in the wild-type mouse, which demonstrate that intravitreal administration of 2’-OMePS-SSO allows selective alteration of Cep290 splicing in retinal cells, including photoreceptors as shown by successful alteration of Abca4 splicing using the same approach. We show that both SSOs and Cep290 skipped mRNA were detectable for at least 1 month and that intravitreal administration of oligonucleotides did not provoke any serious adverse event. These data suggest that intravitreal injections of SSO should be considered to bypass protein truncation resulting from the c.2991+1655A>G mutation as well as other truncating mutations in genes which like CEP290 or ABCA4 have a mRNA size that exceed cargo capacities of US Food and Drug Administration (FDA)-approved adeno-associated virus (AAV)-vectors, thus hampering gene augmentation therapy.http://www.sciencedirect.com/science/article/pii/S216225311630035XCEP290intravitreal injectionLeber congenital amaurosis2′OMePS antisense oligonucleotideretinal dystrophiessplice-switching therapy |
| spellingShingle | Xavier Gérard Isabelle Perrault Arnold Munnich Josseline Kaplan Jean-Michel Rozet Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells Molecular Therapy: Nucleic Acids CEP290 intravitreal injection Leber congenital amaurosis 2′OMePS antisense oligonucleotide retinal dystrophies splice-switching therapy |
| title | Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells |
| title_full | Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells |
| title_fullStr | Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells |
| title_full_unstemmed | Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells |
| title_short | Intravitreal Injection of Splice-switching Oligonucleotides to Manipulate Splicing in Retinal Cells |
| title_sort | intravitreal injection of splice switching oligonucleotides to manipulate splicing in retinal cells |
| topic | CEP290 intravitreal injection Leber congenital amaurosis 2′OMePS antisense oligonucleotide retinal dystrophies splice-switching therapy |
| url | http://www.sciencedirect.com/science/article/pii/S216225311630035X |
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