Cargando…

Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290

Leber congenital amaurosis (LCA) is a rare inherited retinal disorder affecting approximately 1:50,000 people worldwide. So far, mutations in 25 genes have been associated with LCA, with CEP290 (encoding the Centrosomal protein of 290 kDa) being the most frequently mutated gene. The most recurrent L...

Descripción completa

Detalles Bibliográficos
Autores principales: Duijkers, Lonneke, van den Born, L. Ingeborgh, Neidhardt, John, Bax, Nathalie M., Pierrache, Laurence H. M., Klevering, B. Jeroen, Collin, Rob W. J., Garanto, Alejandro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877614/
https://www.ncbi.nlm.nih.gov/pubmed/29518907
http://dx.doi.org/10.3390/ijms19030753
_version_ 1783310732333416448
author Duijkers, Lonneke
van den Born, L. Ingeborgh
Neidhardt, John
Bax, Nathalie M.
Pierrache, Laurence H. M.
Klevering, B. Jeroen
Collin, Rob W. J.
Garanto, Alejandro
author_facet Duijkers, Lonneke
van den Born, L. Ingeborgh
Neidhardt, John
Bax, Nathalie M.
Pierrache, Laurence H. M.
Klevering, B. Jeroen
Collin, Rob W. J.
Garanto, Alejandro
author_sort Duijkers, Lonneke
collection PubMed
description Leber congenital amaurosis (LCA) is a rare inherited retinal disorder affecting approximately 1:50,000 people worldwide. So far, mutations in 25 genes have been associated with LCA, with CEP290 (encoding the Centrosomal protein of 290 kDa) being the most frequently mutated gene. The most recurrent LCA-causing CEP290 mutation, c.2991+1655A>G, causes the insertion of a pseudoexon into a variable proportion of CEP290 transcripts. We previously demonstrated that antisense oligonucleotides (AONs) have a high therapeutic potential for patients homozygously harbouring this mutation, although to date, it is unclear whether rescuing one single allele is enough to restore CEP290 function. Here, we assessed the AON efficacy at RNA, protein and cellular levels in samples that are compound heterozygous for this mutation, together with a protein-truncating mutation in CEP290. We demonstrate that AONs can efficiently restore splicing and increase protein levels. However, due to a high variability in ciliation among the patient-derived cell lines, the efficacy of the AONs was more difficult to assess at the cellular level. This observation points towards the importance of the severity of the second allele and possibly other genetic variants present in each individual. Overall, AONs seem to be a promising tool to treat CEP290-associated LCA, not only in homozygous but also in compound heterozygous carriers of the c.2991+1655A>G variant.
format Online
Article
Text
id pubmed-5877614
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-58776142018-04-09 Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290 Duijkers, Lonneke van den Born, L. Ingeborgh Neidhardt, John Bax, Nathalie M. Pierrache, Laurence H. M. Klevering, B. Jeroen Collin, Rob W. J. Garanto, Alejandro Int J Mol Sci Article Leber congenital amaurosis (LCA) is a rare inherited retinal disorder affecting approximately 1:50,000 people worldwide. So far, mutations in 25 genes have been associated with LCA, with CEP290 (encoding the Centrosomal protein of 290 kDa) being the most frequently mutated gene. The most recurrent LCA-causing CEP290 mutation, c.2991+1655A>G, causes the insertion of a pseudoexon into a variable proportion of CEP290 transcripts. We previously demonstrated that antisense oligonucleotides (AONs) have a high therapeutic potential for patients homozygously harbouring this mutation, although to date, it is unclear whether rescuing one single allele is enough to restore CEP290 function. Here, we assessed the AON efficacy at RNA, protein and cellular levels in samples that are compound heterozygous for this mutation, together with a protein-truncating mutation in CEP290. We demonstrate that AONs can efficiently restore splicing and increase protein levels. However, due to a high variability in ciliation among the patient-derived cell lines, the efficacy of the AONs was more difficult to assess at the cellular level. This observation points towards the importance of the severity of the second allele and possibly other genetic variants present in each individual. Overall, AONs seem to be a promising tool to treat CEP290-associated LCA, not only in homozygous but also in compound heterozygous carriers of the c.2991+1655A>G variant. MDPI 2018-03-07 /pmc/articles/PMC5877614/ /pubmed/29518907 http://dx.doi.org/10.3390/ijms19030753 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Duijkers, Lonneke
van den Born, L. Ingeborgh
Neidhardt, John
Bax, Nathalie M.
Pierrache, Laurence H. M.
Klevering, B. Jeroen
Collin, Rob W. J.
Garanto, Alejandro
Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title_full Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title_fullStr Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title_full_unstemmed Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title_short Antisense Oligonucleotide-Based Splicing Correction in Individuals with Leber Congenital Amaurosis due to Compound Heterozygosity for the c.2991+1655A>G Mutation in CEP290
title_sort antisense oligonucleotide-based splicing correction in individuals with leber congenital amaurosis due to compound heterozygosity for the c.2991+1655a>g mutation in cep290
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877614/
https://www.ncbi.nlm.nih.gov/pubmed/29518907
http://dx.doi.org/10.3390/ijms19030753
work_keys_str_mv AT duijkerslonneke antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT vandenbornlingeborgh antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT neidhardtjohn antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT baxnathaliem antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT pierrachelaurencehm antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT kleveringbjeroen antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT collinrobwj antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290
AT garantoalejandro antisenseoligonucleotidebasedsplicingcorrectioninindividualswithlebercongenitalamaurosisduetocompoundheterozygosityforthec29911655agmutationincep290