DUX4 Transcript Knockdown with Antisense 2′-O-Methoxyethyl Gapmers for the Treatment of Facioscapulohumeral Muscular Dystrophy
DUX4 Transcript Knockdown with Antisense 2′-O-Methoxyethyl Gapmers for the Treatment of Facioscapulohumeral Muscular Dystrophy
复制标题
DOI：
10.1016/j.ymthe.2020.10.010
复制
发表时间：
2021-02-03
影响因子：
12.4
通讯作者：
Yokota, Toshifumi
中科院分区：
文献类型：
--
作者：
Lim, Kenji Rowel Q.;Bittel, Adam;Yokota, Toshifumi
面肩肱骨肌营养不良症（FSHD）是一种常染色体显性遗传病，其特征是肌肉的进行性、不对称弱化，从上半身开始。它是由骨骼肌双同源盒蛋白4基因（DUX4）的异常表达引起的。FSHD目前是无法治愈的。我们建议开发一种利用反义2'- o -甲氧基乙基（2'-MOE）突变子来抑制DUX4 mRNA表达的FSHD治疗方法。在FLExDUX4 FSHD小鼠模型中使用永活的患者来源的肌肉细胞和局部肌内注射，我们发现我们设计的2'-MOE基因片段在体外和体内分别显著降低了DUX4转录水平。此外，在体外，我们观察到dux4激活的下游靶点的表达显著降低，通过RNA测序恢复FSHD特征基因，肌管形态显著改善，脱靶活性最小。这项工作促进了FSHD有前途的候选疗法的发展，并为体内全身治疗研究奠定了基础。
Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant disorder characterized by a progressive, asymmetric weakening of muscles, starting with those in the upper body. It is caused by aberrant expression of the double homeobox protein 4 gene (DUX4) in skeletal muscle. FSHD is currently incurable. We propose to develop a therapy for FSHD using anti sense 2'-O-methoxyethyl (2'-MOE) gapmers, to knock down DUX4 mRNA expression. Using immortalized patient-derived muscle cells and local intramuscular injections in the FLExDUX4 FSHD mouse model, we showed that our designed 2'-MOE gapmers significantly reduced DUX4 transcript levels in vitro and in vivo, respectively. Furthermore, in vitro, we observed significantly reduced expression of DUX4-activated downstream targets, restoration of FSHD signature genes by RNA sequencing, significant improvements in myotube morphology, and minimal off-target activity. This work facilitates the development of a promising candidate therapy for FSHD and lays down the foundation for in vivo systemic treatment studies.