Allele-specific silencing as treatment for gene duplication disorders: proof-of-principle in autosomal dominant leukodystrophy
| Autor: | Giorgio, Elisa, Lorenzati, Martina, Rivetti di Val Cervo, Pia, Brussino, Alessandro, Cernigoj, Manuel, Della Sala, Edoardo, Bartoletti Stella, Anna, Ferrero, Marta, Caiazzo, Massimiliano, Capellari, Sabina, Cortelli, Pietro, Conti, Luciano, Cattaneo, Elena, Buffo, Annalisa, Brusco, Alfredo, Afd Pharmaceutics, Pharmaceutics |
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| Přispěvatelé: | Giorgio E., Lorenzati M., Di Val Cervo P.R., Brussino A., Cernigoj M., Sala E.D., Stella A.B., Ferrero M., Caiazzo M., Capellari S., Cortelli P., Conti L., Cattaneo E., Buffo A., Brusco A., Afd Pharmaceutics, Pharmaceutics, Giorgio, E., Lorenzati, M., Di Val Cervo, P. R., Brussino, A., Cernigoj, M., Sala, E. D., Stella, A. B., Ferrero, M., Caiazzo, M., Capellari, S., Cortelli, P., Conti, L., Cattaneo, E., Buffo, A., Brusco, A. |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
leukodystrophy Small interfering RNA Pelizaeus-Merzbacher Disease Genetic Vectors Single-nucleotide polymorphism Biology 03 medical and health sciences 0302 clinical medicine ADLD RNA interference Gene Duplication medicine Gene silencing Animals Humans Gene Silencing Allele RNA Small Interfering Gene Alleles Cells Cultured Genetics Neurons Lamin Type B Leukodystrophy Lentivirus Genetic Diseases Inborn RNA therapeutics RNA therapeutic Fibroblasts medicine.disease Rats Minor allele frequency LMNB1 030104 developmental biology siRNA Case-Control Studies Neurology (clinical) 030217 neurology & neurosurgery |
| Zdroj: | Brain, 142(7). Oxford University Press |
| ISSN: | 1460-2156 0006-8950 |
| Popis: | Allele-specific silencing by RNA interference (ASP-siRNA) holds promise as a therapeutic strategy for downregulating a single mutant allele with minimal suppression of the corresponding wild-type allele. This approach has been effectively used to target autosomal dominant mutations and single nucleotide polymorphisms linked with aberrantly expanded trinucleotide repeats. Here, we propose ASP-siRNA as a preferable choice to target duplicated disease genes, avoiding potentially harmful excessive downregulation. As a proof-of-concept, we studied autosomal dominant adult-onset demyelinating leukodystrophy (ADLD) due to lamin B1 (LMNB1) duplication, a hereditary, progressive and fatal disorder affecting myelin in the CNS. Using a reporter system, we screened the most efficient ASP-siRNAs preferentially targeting one of the alleles at rs1051644 (average minor allele frequency: 0.45) located in the 3′ untranslated region of the gene. We identified four siRNAs with a high efficacy and allele-specificity, which were tested in ADLD patient-derived fibroblasts. Three of the small interfering RNAs were highly selective for the target allele and restored both LMNB1 mRNA and protein levels close to control levels. Furthermore, small interfering RNA treatment abrogates the ADLD-specific phenotypes in fibroblasts and in two disease-relevant cellular models: murine oligodendrocytes overexpressing human LMNB1, and neurons directly reprogrammed from patients’ fibroblasts. In conclusion, we demonstrated that ASP-silencing by RNA interference is a suitable and promising therapeutic option for ADLD. Moreover, our results have a broad translational value extending to several pathological conditions linked to gene-gain in copy number variations. |
| Databáze: | OpenAIRE |
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