Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice.
Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice.
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DOI：
10.1002/emmm.201202168
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发表时间：
2013-07
影响因子：
11.1
通讯作者：
Carrier, Lucie
中科院分区：
文献类型：
--
作者：
Gedicke-Hornung, Christina;Behrens-Gawlik, Verena;Reischmann, Silke;Geertz, Birgit;Stimpel, Doreen;Weinberger, Florian;Schlossarek, Saskia;Precigout, Guillaume;Braren, Ingke;Eschenhagen, Thomas;Mearini, Giulia;Lorain, Stephanie;Voit, Thomas;Dreyfus, Patrick A.;Garcia, Luis;Carrier, Lucie
关键词：
反义寡核苷酸介导的外显子跳跃是一种很有前途的治疗遗传性疾病的方法，但尚未评估心脏疾病。我们研究了在Mybpc 3靶向基因敲入（KI）的肥厚型心肌病（HCM）小鼠模型中病毒介导的AON转移的可行性和有效性。KI小鼠在外显子6中携带纯合的G>A转换，这导致三种不同的异常mRNA。我们在野生型和KI小鼠中鉴定了外显子5-6缺失的替代变体（Var-4）。为了增强其表达并抑制异常mRNA，我们设计了AON-5和AON-6，其掩蔽外显子5和6中的剪接增强子基序。将AON插入修饰的U 7小核RNA中并包装在腺相关病毒（AAV-U 7-AON-5+6）中。转导心肌细胞或全身施用AAV-U 7-AON-5+6增加了Var-4 mRNA/蛋白水平并减少了异常mRNA。注射新生KI小鼠消除心功能不全，并防止左心室肥大。虽然治疗效果是短暂的，因此需要在较长时间内保持优化，但这项概念验证研究为HCM的因果治疗铺平了道路。
Exon skipping mediated by antisense oligoribonucleotides (AON) is a promising therapeutic approach for genetic disorders, but has not yet been evaluated for cardiac diseases. We investigated the feasibility and efficacy of viral-mediated AON transfer in a Mybpc3-targeted knock-in (KI) mouse model of hypertrophic cardiomyopathy (HCM). KI mice carry a homozygous G>A transition in exon 6, which results in three different aberrant mRNAs. We identified an alternative variant (Var-4) deleted of exons 5–6 in wild-type and KI mice. To enhance its expression and suppress aberrant mRNAs we designed AON-5 and AON-6 that mask splicing enhancer motifs in exons 5 and 6. AONs were inserted into modified U7 small nuclear RNA and packaged in adeno-associated virus (AAV-U7-AON-5+6). Transduction of cardiac myocytes or systemic administration of AAV-U7-AON-5+6 increased Var-4 mRNA/protein levels and reduced aberrant mRNAs. Injection of newborn KI mice abolished cardiac dysfunction and prevented left ventricular hypertrophy. Although the therapeutic effect was transient and therefore requires optimization to be maintained over an extended period, this proof-of-concept study paves the way towards a causal therapy of HCM.
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影响因子：
14.9
作者：
Cartegni, L;Wang, JH;Krainer, AR
通讯作者：
Krainer, AR
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Heemskerk, Hans A.;de Winter, Christa L.;Aartsma-Rus, Annemieke
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通讯作者：
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作者：
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通讯作者：
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