Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.18/3429
Título: Functional analysis of splicing mutations in the IDS gene and the use of antisense oligonucleotides to exploit an alternative therapy for MPS II
Autor: Matos, Liliana
Gonçalves, Vânia
Pinto, Eugénia
Laranjeira, Francisco
Prata, Maria João
Jordan, Peter
Desviat, Lourdes R.
Perez, Belén
Alves, Sandra
Palavras-chave: Doenças Genéticas
Lysosomal Storage Disorders
Splicing Regulation
Mutation
Antisense Therapy
IDS gene
Data: Dez-2015
Editora: Elsevier
Citação: Biochim Biophys Acta. 2015 Dec;1852(12):2712-21. doi: 10.1016/j.bbadis.2015.09.011. Epub 2015 Sep 25
Resumo: Mucopolysaccharidosis II is a lysosomal storage disorder caused by mutations in the IDS gene, including exonic alterations associated with aberrant splicing. In the present work, cell-based splicing assays were performed to study the effects of two splicing mutations in exon 3 of IDS, i.e., c.241C>T and c.257C>T, whose presence activates a cryptic splice site in exon 3 and one in exon 8, i.e., c.1122C>T that despite being a synonymous mutation is responsible for the creation of a new splice site in exon 8 leading to a transcript shorter than usual. Mutant minigene analysis and overexpression assays revealed that SRSF2 and hnRNP E1 might be involved in the use and repression of the constitutive 3' splice site of exon 3 respectively. For the c.1122C>T the use of antisense therapy to correct the splicing defect was explored, but transfection of patient fibroblasts with antisense morpholino oligonucleotides (n=3) and a locked nucleic acid failed to abolish the abnormal transcript; indeed, it resulted in the appearance of yet another aberrant splicing product. Interestingly, the oligonucleotides transfection in control fibroblasts led to the appearance of the aberrant transcript observed in patients' cells after treatment, which shows that the oligonucleotides are masking an important cis-acting element for 5' splice site regulation of exon 8. These results highlight the importance of functional studies for understanding the pathogenic consequences of mis-splicing and highlight the difficulty in developing antisense therapies involving gene regions under complex splicing regulation.
Peer review: yes
URI: http://hdl.handle.net/10400.18/3429
DOI: 10.1016/j.bbadis.2015.09.011
ISSN: 0006-3002
Versão do Editor: http://www.sciencedirect.com/science/article/pii/S0925443915002835
Aparece nas colecções:DGH - Artigos em revistas internacionais

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