Designed U7 snRNAs inhibit DUX4 expression and improve FSHD-associated outcomes in DUX4 overexpressing cells and FSHD patient myotubes.
Designed U7 snRNAs inhibit DUX4 expression and improve FSHD-associated outcomes in DUX4 overexpressing cells and FSHD patient myotubes.
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DOI：
10.1016/j.omtn.2020.12.004
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发表时间：
2021-03-05
期刊：
影响因子：
--
通讯作者：
Harper SQ
中科院分区：
文献类型：
--
作者：
Rashnonejad A;Amini-Chermahini G;Taylor NK;Wein N;Harper SQ
面肩肱型肌营养不良症 (FSHD) 是由肌肉中 DUX4 基因去抑制的表观遗传变化引起的。全长 DUX4 蛋白会导致细胞死亡和肌肉毒性，因此我们假设 FSHD 疗法应集中于抑制全长 DUX4 表达。在这项研究中，我们开发了一种使用 U7-小核 RNA (snRNA) 反义表达盒（称为 U7-asDUX4）来实现 DUX4 抑制的策略。这些非编码 RNA 旨在通过掩蔽 DUX4 起始密码子、剪接位点或多腺苷酸化信号来抑制全长 DUX4 前 mRNA 的产生或成熟。在此过程中，U7-asDUX4 snRNA 的运作方式与反义寡核苷酸类似。然而，与寡核苷酸相比，寡核苷酸受到肌肉吸收不良和需要终生重复给药的限制，U7-asDUX4 snRNA 可以包装在促肌基因治疗载体中，并且在体内递送至有丝分裂后细胞时可能只需要单次给药。我们测试了几种 U7-asDUX4，它们可在体外降低 DUX4 表达并改善 DUX4 相关结果。通过 U7-snRNA 抑制 DUX4 表达可能是 FSHD 的一种新的前瞻性基因治疗方法，或者与其他策略（如 RNAi 治疗）联合使用，以最大限度地沉默 FSHD 个体中的 DUX4。面肩肱型肌营养不良症 (FSHD) 的治疗旨在抑制导致该疾病的有毒 DUX4 基因。这项研究描述了一种在 mRNA 水平抑制 DUX4 产生的新方法，即使用设计的 U7 反义 RNA 表达盒在空间上阻碍 DUX4 mRNA 的成熟和翻译。
Facioscapulohumeral muscular dystrophy (FSHD) arises from epigenetic changes that de-repress the DUX4 gene in muscle. The full-length DUX4 protein causes cell death and muscle toxicity, and therefore we hypothesize that FSHD therapies should center on inhibiting full-length DUX4 expression. In this study, we developed a strategy to accomplish DUX4 inhibition using U7-small nuclear RNA (snRNA) antisense expression cassettes (called U7-asDUX4). These non-coding RNAs were designed to inhibit production or maturation of the full-length DUX4 pre-mRNA by masking the DUX4 start codon, splice sites, or polyadenylation signal. In so doing, U7-asDUX4 snRNAs operate similarly to antisense oligonucleotides. However, in contrast to oligonucleotides, which are limited by poor uptake in muscle and a requirement for lifelong repeated dosing, U7-asDUX4 snRNAs can be packaged within myotropic gene therapy vectors and may require only a single administration when delivered to post-mitotic cells in vivo. We tested several U7-asDUX4s that reduced DUX4 expression in vitro and improved DUX4-associated outcomes. Inhibition of DUX4 expression via U7-snRNAs could be a new prospective gene therapy approach for FSHD or be used in combination with other strategies, like RNAi therapy, to maximize DUX4 silencing in individuals with FSHD. Treatments for facioscapulohumeral muscular dystrophy (FSHD) are designed to suppress the toxic DUX4 gene that underlies the disorder. This study describes a novel approach to inhibit DUX4 production at the mRNA level by using designed U7-antisense RNA expression cassettes to sterically hinder maturation and translation of DUX4 mRNA.
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影响因子：
12.4
作者：
Heemskerk, Hans;de Winter, Christa;van Kuik, Petra;Heuvelmans, Niki;Sabatelli, Patrizia;Rimessi, Paola;Braghetta, Paola;van Ommen, Gert-Jan B.;de Kimpe, Sjef;Ferlini, Alessandra;Aartsma-Rus, Annemieke;van Deutekom, Judith C. T.
通讯作者：
van Deutekom, Judith C. T.
DOI：
10.1038/mt.2016.111
发表时间：
2016-08
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
通讯作者：
--
影响因子：
17.1
作者：
Lek, Angela;Zhang, Yuanfan;Kunkel, Louis M.
通讯作者：
Kunkel, Louis M.
影响因子：
12.4
作者：
Goyenvalle, Aurelie;Wright, Jordan;Davies, Kay E.
通讯作者：
Davies, Kay E.
影响因子：
3.5
作者：
Gabriëls, J;Beckers, MC;Belayew, A
通讯作者：
Belayew, A