In vitro and in vivo rescue of aberrant splicing in CEP290-associated LCA by antisense oligonucleotide delivery.
Leber congenital amaurosis (LCA) is a severe disorder resulting in visual impairment usually starting in the first year of life. The most frequent genetic cause of LCA is an intronic mutation in CEP290 (c.2991 + 1655A > G) that creates a cryptic splice donor site resulting in the insertion of a pseudoexon (exon X) into CEP290 mRNA. Previously, we showed that naked antisense oligonucleotides (AONs) effectively restored normal CEP290 splicing in patient-derived lymphoblastoid cells. We here explore the therapeutic potential of naked and adeno-associated virus (AAV)-packaged AONs in vitro and in vivo In both cases, AON delivery fully restored CEP290 pre-mRNA splicing, significantly increased CEP290 protein levels and rescued a ciliary phenotype present in patient-derived fibroblast cells. Moreover, administration of naked and AAV-packaged AONs to the retina of a humanized mutant Cep290 mouse model, carrying the intronic mutation, showed a statistically significant reduction of exon X-containing Cep290 transcripts, without compromising the retinal structure. Together, our data highlight the tremendous therapeutic prospective of AONs for the treatment of not only CEP290-associated LCA but potentially many other subtypes of retinal dystrophy caused by splicing mutations.
Department of Human Genetics.|Donders Center for NeurosciencesRadboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
Alejandro
A
Garanto
F.M. Kirby Center for Molecular Ophthalmology and Center for Advanced Retinal and Ophthalmic Therapeutics, Scheie Eye Institute, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Daniel C
DC
Chung
Department of Human Genetics.
Lonneke
L
Duijkers
Department of Human Genetics.|Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
Julio C
JC
Corral-Serrano
Department of Human Genetics.|Donders Center for NeurosciencesRadboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
Muriël
M
Messchaert
Grousbeck Gene Therapy Center, Department of Ophthalmology, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA 02114, USA.
Ru
R
Xiao
F.M. Kirby Center for Molecular Ophthalmology and Center for Advanced Retinal and Ophthalmic Therapeutics, Scheie Eye Institute, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Jean
J
Bennett
Grousbeck Gene Therapy Center, Department of Ophthalmology, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA 02114, USA.
Luk H
LH
Vandenberghe
Department of Human Genetics rob.collin@radboudumc.nl.|Donders Center for NeurosciencesRadboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
Rob W J
RW
Collin
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000627:therapeutic use; D001766:Blindness / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D005091:Exons / Q000235:genetics; D015316:Genetic Therapy; D006801:Humans; D057130:Leber Congenital Amaurosis / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D051379:Mice; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics* / Q000627:therapeutic use; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics; D010641:Phenotype; D022821:RNA Splice Sites / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D012160:Retina / Q000187:drug effects / Q000473:pathology
D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C493903:Cep290 protein, human; D003598:Cytoskeletal Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D022821:RNA Splice Sites