High-throughput screening identifies inhibitors of DUX4-induced myoblast toxicity.
High-throughput screening identifies inhibitors of DUX4-induced myoblast toxicity.
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DOI：
10.1186/2044-5040-4-4
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发表时间：
2014-02-01
期刊：
影响因子：
4.9
通讯作者：
Kyba M
中科院分区：
文献类型：
--
作者：
Bosnakovski D;Choi SH;Strasser JM;Toso EA;Walters MA;Kyba M
面肩肱型肌营养不良症（FSHD）是由4号染色体上D4 Z4大卫星重复位点的表观遗传学改变引起的，导致DUX 4蛋白的不适当表达。因此，DUX 4蛋白是治疗干预的主要分子靶标。我们已经开发了一种基于DUX 4在C2 C12成肌细胞中过表达时的毒性的高通量筛选，并从44，000种小的药物样分子中鉴定了DUX 4诱导毒性的抑制剂。然后对总共1，280个命中物进行针对由3 T3成纤维细胞表达的DUX 4的活性、针对用于条件性表达DUX 4的tet-on系统的活性的缺乏以及对细胞增殖速率的潜在影响的二次筛选。这使我们能够定义一组52种化合物作为探针来识别DUX 4活性的基本途径。我们测试了这些化合物保护野生型细胞免受其他类型细胞死亡诱导损伤的能力。值得注意的是，我们发现60%的DUX 4毒性抑制剂也可以保护细胞免受叔丁基过氧化氢（一种氧化应激诱导化合物）的影响。化合物不能防止半胱天冬酶激活、DNA损伤、蛋白质错误折叠或ER应激诱导的死亡。令人鼓舞的是，这些化合物中的许多也对人细胞中的DUX 4表达具有保护作用。这些数据表明，氧化应激是DUX 4引起的毒性在该系统中介导的主要途径，我们推测，增强氧化应激反应途径可能在FSHD中具有临床益处。
Facioscapulohumeral muscular dystrophy (FSHD) is caused by epigenetic alterations at the D4Z4 macrosatellite repeat locus on chromosome 4, resulting in inappropriate expression of the DUX4 protein. The DUX4 protein is therefore the primary molecular target for therapeutic intervention. We have developed a high-throughput screen based on the toxicity of DUX4 when overexpressed in C2C12 myoblasts, and identified inhibitors of DUX4-induced toxicity from within a diverse set of 44,000 small, drug-like molecules. A total of 1,280 hits were then subjected to secondary screening for activity against DUX4 expressed by 3T3 fibroblasts, for absence of activity against the tet-on system used to conditionally express DUX4, and for potential effects on cellular proliferation rate. This allowed us to define a panel of 52 compounds to use as probes to identify essential pathways of DUX4 activity. We tested these compounds for their ability to protect wild-type cells from other types of cell death-inducing insults. Remarkably, we found that 60% of the DUX4 toxicity inhibitors that we identified also protected cells from tert-butyl hydrogen peroxide, an oxidative stress-inducing compound. Compounds did not protect against death induced by caspase activation, DNA damage, protein misfolding, or ER stress. Encouragingly, many of these compounds are also protective against DUX4 expression in human cells. These data suggest that oxidative stress is a dominant pathway through which DUX4-provoked toxicity is mediated in this system, and we speculate that enhancing the oxidative stress response pathway might be clinically beneficial in FSHD.
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DOI：
10.1093/brain/awp236
发表时间：
2009-11
期刊：
Brain : a journal of neurology
影响因子：
--
作者：
Norwood FL;Harling C;Chinnery PF;Eagle M;Bushby K;Straub V
通讯作者：
Straub V
影响因子：
3.5
作者：
Winokur, ST;Chen, YW;Flanigan, KM
通讯作者：
Flanigan, KM
影响因子：
2.8
作者：
Winokur, ST;Barrett, K;Figlewicz, DA
通讯作者：
Figlewicz, DA
影响因子：
3.5
作者：
Gabriëls, J;Beckers, MC;Belayew, A
通讯作者：
Belayew, A
DOI：
10.1073/pnas.0708659104
发表时间：
2007-11-13
影响因子：
11.1
作者：
Dixit, Manjusha;Ansseau, Eugenie;Chen, Yi-Wen
通讯作者：
Chen, Yi-Wen