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Suppression of galactocerebrosidase premature termination codon and rescue of galactocerebrosidase activity in twitcher cells
Author(s) -
Luddi Alice,
Crifasi Laura,
Capaldo Angela,
Piomboni Paola,
CostantinoCeccarini Elvira
Publication year - 2016
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.23790
Subject(s) - krabbe disease , nonsense mutation , nonsense mediated decay , heterozygote advantage , biology , stop codon , mutation , compound heterozygosity , nonsense , genetics , microbiology and biotechnology , gene , leukodystrophy , genotype , medicine , disease , rna , missense mutation , rna splicing
Krabbe's disease (KD) is a degenerative lysosomal storage disease resulting from deficiency of β‐galactocerebrosidase activity. Over 100 mutations are known to cause the disease, and these usually occur in compound heterozygote patterns. In affected patients, nonsense mutations leading to a nonfunctional enzyme are often found associated with other mutations. The twitcher mouse is a naturally occurring model of KD, containing in β‐galactocerebrosidase a premature stop codon, W339X. Recent studies have shown that selected compounds may induce the ribosomal bypass of premature stop codons without affecting the normal termination codons. The rescue of β‐galactocerebrosidase activity induced by treatment with premature termination codon (PTC) 124, a well‐characterized compound known to induce ribosomal read‐through, was investigated on oligodendrocytes prepared from twitcher mice and on human fibroblasts from patients bearing nonsense mutations. The effectiveness of the nonsense‐mediated mRNA decay (NMD) inhibitor 1 (NMDI1), a newly identified inhibitor of NMD, was also tested. Incubation of these cell lines with PTC124 and NMDI1 increased the levels of mRNA and rescued galactocerebrosidase enzymatic activity in a dose‐dependent manner. The low but sustained expression of β‐galactocerebrosidase in oligodendrocytes was sufficient to improve the morphology of the differentiated cells. Our in vitro approach provides the basis for further investigation of ribosomal read‐through as an alternative therapeutic strategy to ameliorate the quality of life in selected KD patients. © 2016 Wiley Periodicals, Inc.