Allele-specific silencing of mutant huntingtin and ataxin-3 genes by targeting expanded CAG repeats in mRNAs

Hu, Jiaxin; Matsui, Masayuki; Gagnon, Keith T.; Schwartz, Jacob C.; Gabillet, Sylvie; Arar, Khalil; Wu, Jun; Bezprozvanny, Ilya; Corey, David R.

doi:10.1038/nbt.1539

Cited by 221 publications

(277 citation statements)

References 34 publications

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“…2′-OMe ASOs have also been shown to selectively target expanded CUG repeats in DM1 myoblasts (13). Another study showed that ASOs used to inhibit translation of CAG expansions in Huntington disease only affected the expanded huntingtin allele but not the wild-type allele (26). This provides evidence that the ability to preferentially target expanded repeat sequence may be common to other ASO therapies.…”

Section: Discussionmentioning

confidence: 76%

RNase H-mediated degradation of toxic RNA in myotonic dystrophy type 1

Lee

Bennett

Cooper

2012

Proc. Natl. Acad. Sci. U.S.A.

143

134

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Myotonic dystrophy type 1 (DM1) is an RNA-dominant disease caused by abnormal transcripts containing expanded CUG repeats. The CUG transcripts aggregate in the nucleus to form RNA foci and lead to nuclear depletion of Muscleblind-like 1 (MBNL1) and stabilized expression of CUGBP Elav like family 1 (CELF1), both of which are splicing regulatory proteins. The imbalance of these proteins results in misregulation of alternative splicing and neuromuscular abnormalities. Here, we report the use of antisense oligonucleotides (ASOs) as a therapeutic approach to target the pathogenic RNA in DM1. We designed chimeric ASOs, termed gapmers, containing modified nucleic acid residues to induce RNase H-mediated degradation of CUG-repeat transcripts. The gapmers selectively knockdown expanded CUG transcripts and are sufficient to disrupt RNA foci both in cell culture and mouse models for DM1. Furthermore, combination of gapmers with morpholino ASOs that help release binding of MBNL1 to the toxic RNA can potentially enhance the knockdown effect. Additional optimization will be required for systemic delivery; however, our study provides an alternative strategy for the use of ASOs in DM1 therapy.microsatellite expansion | muscular dystrophy | phosphorothioate | gapmer

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Section: Discussionmentioning

confidence: 76%

RNase H-mediated degradation of toxic RNA in myotonic dystrophy type 1

Lee

Bennett

Cooper

2012

Proc. Natl. Acad. Sci. U.S.A.

143

134

View full text Add to dashboard Cite

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“…6,11,21 However, only a few studies have assessed silencing in patient-derived cells, which exhibit physiologically relevant levels of endogenous expression. 3,[22][23][24][25] We sought to address whether polyQ patient-derived fibroblasts might be a useful model for assessing therapeutic strategies, such as comparing allele-specific and non-allele-specific silencing approaches in genetically accurate cells. All the polyQ patient-derived cell line studies to date 3,[22][23][24][25][26] have focused on the development of allelespecific silencing approaches targeting SNPs linked to the mutation and/or the mutation itself, and show increasingly promising results.…”

Section: Discussionmentioning

confidence: 99%

Allele-specific silencing of mutant Ataxin-7 in SCA7 patient-derived fibroblasts

et al. 2014

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Polyglutamine (polyQ) disorders are inherited neurodegenerative conditions defined by a common pathogenic CAG repeat expansion leading to a toxic gain-of-function of the mutant protein. Consequences of this toxicity include activation of heatshock proteins (HSPs), impairment of the ubiquitin-proteasome pathway and transcriptional dysregulation. Several studies in animal models have shown that reducing levels of toxic protein using small RNAs would be an ideal therapeutic approach for such disorders, including spinocerebellar ataxia-7 (SCA7). However, testing such RNA interference (RNAi) effectors in genetically appropriate patient cell lines with a disease-relevant phenotype has yet to be explored. Here, we have used primary adult dermal fibroblasts from SCA7 patients and controls to assess the endogenous allele-specific silencing of ataxin-7 by two distinct siRNAs. We further identified altered expression of two disease-relevant transcripts in SCA7 patient cells: a twofold increase in levels of the HSP DNAJA1 and a twofold decrease in levels of the de-ubiquitinating enzyme, UCHL1. After siRNA treatment, the expression of both genes was restored towards normal levels. To our knowledge, this is the first time that allelespecific silencing of mutant ataxin-7, targeting a common SNP, has been demonstrated in patient cells. These findings highlight the advantage of an allele-specific RNAi-based therapeutic approach, and indicate the value of primary patient-derived cells as useful models for mechanistic studies and for measuring efficacy of RNAi effectors on a patient-to-patient basis in the polyQ diseases.

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“…5). Given this, the development of nucleic acid therapy by noncoding small RNAs could be an ideal approach to selectively silence mutant htt [115,116]. Harper et al [117] showed that RNA interference using adeno-associated virus-small hairpin RNA (shRNA) improves motor deficits and neuropathological phenotypes in a transgenic (N171-82Q) mouse model of HD.…”

Section: Rna Interference and Noncoding Small Rnasmentioning

confidence: 99%

Epigenetic Mechanisms of Neurodegeneration in Huntington's Disease

et al. 2013

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Huntington's disease (HD) is an incurable and fatal hereditary neurodegenerative disorder of mid-life onset characterized by chorea, emotional distress, and progressive cognitive decline. HD is caused by an expansion of CAG repeats coding for glutamine (Q) in exon 1 of the huntingtin gene. Recent studies suggest that epigenetic modifications may play a key role in HD pathogenesis. Alterations of the epigenetic "histone code" lead to chromatin remodeling and deregulation of neuronal gene transcription that are prominently linked to HD pathogenesis. Furthermore, specific noncoding RNAs and microRNAs are associated with neuronal damage in HD. In this review, we discuss how DNA methylation, posttranslational modifications of histone, and noncoding RNA function are affected and involved in HD pathogenesis. In addition, we summarize the therapeutic effects of histone deacetylase inhibitors and DNA binding drugs on epigenetic modifications and neuropathological sequelae in HD. Our understanding of the role of these epigenetic mechanisms may lead to the identification of novel biological markers and new therapeutic targets to treat HD.

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Allele-specific silencing of mutant huntingtin and ataxin-3 genes by targeting expanded CAG repeats in mRNAs

Cited by 221 publications

References 34 publications

RNase H-mediated degradation of toxic RNA in myotonic dystrophy type 1

RNase H-mediated degradation of toxic RNA in myotonic dystrophy type 1

Allele-specific silencing of mutant Ataxin-7 in SCA7 patient-derived fibroblasts

Epigenetic Mechanisms of Neurodegeneration in Huntington's Disease

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