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Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts
Published on: May 11, 2018
In vitro antisense therapeutics for a deep intronic mutation causing Neurofibromatosis type 2
Elisabeth Castellanos1, Imma Rosas, Ares Solanes
1Hereditary Cancer Program, Institute of Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Spain.
European Journal of Human Genetics : EJHG
|November 29, 2012
Summary
Neurofibromatosis type 2 (NF2) is a genetic disorder. A novel deep intronic mutation causing severe NF2 was corrected in vitro using antisense morpholino oligomers, restoring normal splicing and protein levels.
Area of Science:
- Genetics
- Molecular Biology
- Oncology
Background:
- Neurofibromatosis type 2 (NF2) is an autosomal-dominant disorder affecting approximately 1:33,000 newborns.
- NF2 is characterized by nervous system tumors and ocular abnormalities, with 85% of germline mutations being point mutations.
- Approximately 25% of NF2 point mutations impact mRNA splicing, leading to variable disease severity.
Observation:
- A patient with severe NF2 phenotype was identified with a deep intronic mutation (g. 74409T>A, NG_009057.1).
- This mutation caused the insertion of a cryptic exon into the mature mRNA, resulting in a truncated merlin protein (p.Pro482Profs*39).
Findings:
- A mutation-specific antisense phosphorodiamidate morpholino oligomer (ASO) was designed and tested in vitro.
- The ASO effectively restored normal NF2 splicing in patient-derived fibroblasts.
- Morpholino treatment recovered merlin protein levels, reduced fibroblast proliferation, and restored cytoskeleton organization.
Implications:
- This study presents the first NF2 case caused by a deep intronic mutation with an in vitro tested antisense therapeutic approach.
- These findings suggest the potential for in vivo application of ASO therapy for deep intronic mutations causing NF2.
- This approach may offer a therapeutic strategy for a subset of NF2 patients with splicing defects.
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