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Targeting nuclear RNA for in vivo correction of myotonic dystrophy.

Wheeler TM, Leger AJ, Pandey SK, MacLeod AR, Nakamori M, Cheng SH, Wentworth BM, Bennett CF, Thornton CA - Nature (2012)

Bottom Line: The effect was sustained for up to 1 year after treatment was discontinued.Systemically administered ASOs were also effective for muscle knockdown of Malat1, a long non-coding RNA (lncRNA) that is retained in the nucleus.These results provide a general strategy to correct RNA gain-of-function effects and to modulate the expression of expanded repeats, lncRNAs and other transcripts with prolonged nuclear residence.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of Rochester, 601 Elmwood Avenue, Rochester, New York 14642, USA.

ABSTRACT
Antisense oligonucleotides (ASOs) hold promise for gene-specific knockdown in diseases that involve RNA or protein gain-of-function effects. In the hereditary degenerative disease myotonic dystrophy type 1 (DM1), transcripts from the mutant allele contain an expanded CUG repeat and are retained in the nucleus. The mutant RNA exerts a toxic gain-of-function effect, making it an appropriate target for therapeutic ASOs. However, despite improvements in ASO chemistry and design, systemic use of ASOs is limited because uptake in many tissues, including skeletal and cardiac muscle, is not sufficient to silence target messenger RNAs. Here we show that nuclear-retained transcripts containing expanded CUG (CUG(exp)) repeats are unusually sensitive to antisense silencing. In a transgenic mouse model of DM1, systemic administration of ASOs caused a rapid knockdown of CUG(exp) RNA in skeletal muscle, correcting the physiological, histopathologic and transcriptomic features of the disease. The effect was sustained for up to 1 year after treatment was discontinued. Systemically administered ASOs were also effective for muscle knockdown of Malat1, a long non-coding RNA (lncRNA) that is retained in the nucleus. These results provide a general strategy to correct RNA gain-of-function effects and to modulate the expression of expanded repeats, lncRNAs and other transcripts with prolonged nuclear residence.

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Transcriptomic effects of ASOsMicroarray analysis of gene expression in quadriceps muscle (n = 4 mice per group). a, Principle component analysis shows segregation of HSALR (saline) away from wild-type mice in widely separated clusters. ASOs caused HSALR transgenic mice to cluster nearer to wild-type mice (25 mg/kg biweekly for 4 wks). b, Among transcripts upregulated in HSALR vs. wild-type mice (saline), > 85% showed complete or partial return to normal expression after treatment with ASOs.
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Figure 2: Transcriptomic effects of ASOsMicroarray analysis of gene expression in quadriceps muscle (n = 4 mice per group). a, Principle component analysis shows segregation of HSALR (saline) away from wild-type mice in widely separated clusters. ASOs caused HSALR transgenic mice to cluster nearer to wild-type mice (25 mg/kg biweekly for 4 wks). b, Among transcripts upregulated in HSALR vs. wild-type mice (saline), > 85% showed complete or partial return to normal expression after treatment with ASOs.

Mentions: In addition to splicing defects, expression of CUGexp RNA or ablation of Mbnl1 causes extensive remodeling of the muscle transcriptome16, 17, 23. We used microarrays to examine transcriptomic effects of ASOs. Principle component analysis showed that gene expression in ASO-treated HSALR mice was shifted towards wild-type mice, indicating an overall trend for transcriptome normalization (Fig. 2a). Among transcripts that were up- or down-regulated in HSALR muscle, > 85% were normalized or partially corrected by ASOs, without evidence for off-target effects (Fig. 2b; Supplementary Fig 7; Supplementary Table 2). These results confirm that ASOs caused an overall improvement of the muscle transcriptome.


Targeting nuclear RNA for in vivo correction of myotonic dystrophy.

Wheeler TM, Leger AJ, Pandey SK, MacLeod AR, Nakamori M, Cheng SH, Wentworth BM, Bennett CF, Thornton CA - Nature (2012)

Transcriptomic effects of ASOsMicroarray analysis of gene expression in quadriceps muscle (n = 4 mice per group). a, Principle component analysis shows segregation of HSALR (saline) away from wild-type mice in widely separated clusters. ASOs caused HSALR transgenic mice to cluster nearer to wild-type mice (25 mg/kg biweekly for 4 wks). b, Among transcripts upregulated in HSALR vs. wild-type mice (saline), > 85% showed complete or partial return to normal expression after treatment with ASOs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4221572&req=5

Figure 2: Transcriptomic effects of ASOsMicroarray analysis of gene expression in quadriceps muscle (n = 4 mice per group). a, Principle component analysis shows segregation of HSALR (saline) away from wild-type mice in widely separated clusters. ASOs caused HSALR transgenic mice to cluster nearer to wild-type mice (25 mg/kg biweekly for 4 wks). b, Among transcripts upregulated in HSALR vs. wild-type mice (saline), > 85% showed complete or partial return to normal expression after treatment with ASOs.
Mentions: In addition to splicing defects, expression of CUGexp RNA or ablation of Mbnl1 causes extensive remodeling of the muscle transcriptome16, 17, 23. We used microarrays to examine transcriptomic effects of ASOs. Principle component analysis showed that gene expression in ASO-treated HSALR mice was shifted towards wild-type mice, indicating an overall trend for transcriptome normalization (Fig. 2a). Among transcripts that were up- or down-regulated in HSALR muscle, > 85% were normalized or partially corrected by ASOs, without evidence for off-target effects (Fig. 2b; Supplementary Fig 7; Supplementary Table 2). These results confirm that ASOs caused an overall improvement of the muscle transcriptome.

Bottom Line: The effect was sustained for up to 1 year after treatment was discontinued.Systemically administered ASOs were also effective for muscle knockdown of Malat1, a long non-coding RNA (lncRNA) that is retained in the nucleus.These results provide a general strategy to correct RNA gain-of-function effects and to modulate the expression of expanded repeats, lncRNAs and other transcripts with prolonged nuclear residence.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of Rochester, 601 Elmwood Avenue, Rochester, New York 14642, USA.

ABSTRACT
Antisense oligonucleotides (ASOs) hold promise for gene-specific knockdown in diseases that involve RNA or protein gain-of-function effects. In the hereditary degenerative disease myotonic dystrophy type 1 (DM1), transcripts from the mutant allele contain an expanded CUG repeat and are retained in the nucleus. The mutant RNA exerts a toxic gain-of-function effect, making it an appropriate target for therapeutic ASOs. However, despite improvements in ASO chemistry and design, systemic use of ASOs is limited because uptake in many tissues, including skeletal and cardiac muscle, is not sufficient to silence target messenger RNAs. Here we show that nuclear-retained transcripts containing expanded CUG (CUG(exp)) repeats are unusually sensitive to antisense silencing. In a transgenic mouse model of DM1, systemic administration of ASOs caused a rapid knockdown of CUG(exp) RNA in skeletal muscle, correcting the physiological, histopathologic and transcriptomic features of the disease. The effect was sustained for up to 1 year after treatment was discontinued. Systemically administered ASOs were also effective for muscle knockdown of Malat1, a long non-coding RNA (lncRNA) that is retained in the nucleus. These results provide a general strategy to correct RNA gain-of-function effects and to modulate the expression of expanded repeats, lncRNAs and other transcripts with prolonged nuclear residence.

Show MeSH
Related in: MedlinePlus