Therapeutic strategies based on modified U1 snRNAs and chaperones for Sanfilippo C splicing mutations

Orphanet Journal of Rare Diseases

Lluïsa Vilageliu⁶  Sandra Alves³  Daniel Grinberg⁶  Alexey V Pshezhetsky⁷  Belén Pérez⁴  Lourdes R Desviat⁴  Peter Jordan²  Maria João Prata⁵  Yoo Choi⁷  Larbi Dridi⁷  Isaac Canals⁶  Liliana Matos¹ 

[1] Department of Biology, Faculty of Sciences, Porto, Portugal;Department of Human Genetics, Research and Development Unit, INSA, Lisbon, Portugal;Department of Human Genetics, Research and Development Unit, INSA, Porto, Portugal;Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, UAM-CSIC, Universidad Autónoma de Madrid, Madrid, Spain;IPATIMUP, Porto, Portugal;Institut de Biomedicina de la Universitat de Barcelona (IBUB), Barcelona, Spain;Department of Medical Genetics, Sainte-Justine University Hospital Centre, University of Montreal, Montreal, Canada

关键词: Lysosomal storage disorder;    Sanfilippo C syndrome;    Glucosamine;    Modified U1 snRNAs;    Splicing mutations;   

Others  :  1228425
DOI  :  10.1186/s13023-014-0180-y

 received in 2014-08-13, accepted in 2014-11-04,  发布年份 2014

【 摘 要 】

Background

Mutations affecting RNA splicing represent more than 20% of the mutant alleles in Sanfilippo syndrome type C, a rare lysosomal storage disorder that causes severe neurodegeneration. Many of these mutations are localized in the conserved donor or acceptor splice sites, while few are found in the nearby nucleotides.

Methods

In this study we tested several therapeutic approaches specifically designed for different splicing mutations depending on how the mutations affect mRNA processing. For three mutations that affect the donor site (c.234 + 1G > A, c.633 + 1G > A and c.1542 + 4dupA), different modified U1 snRNAs recognizing the mutated donor sites, have been developed in an attempt to rescue the normal splicing process. For another mutation that affects an acceptor splice site (c.372-2A > G) and gives rise to a protein lacking four amino acids, a competitive inhibitor of the HGSNAT protein, glucosamine, was tested as a pharmacological chaperone to correct the aberrant folding and to restore the normal trafficking of the protein to the lysosome.

Results

Partial correction of c.234 + 1G > A mutation was achieved with a modified U1 snRNA that completely matches the splice donor site suggesting that these molecules may have a therapeutic potential for some splicing mutations. Furthermore, the importance of the splice site sequence context is highlighted as a key factor in the success of this type of therapy. Additionally, glucosamine treatment resulted in an increase in the enzymatic activity, indicating a partial recovery of the correct folding.

Conclusions

We have assayed two therapeutic strategies for different splicing mutations with promising results for the future applications.

【 授权许可】

   
2014 Matos et al.; licensee BioMed Central.

【 图 表 】

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