Duchenne muscular dystrophy (DMD) is a severe neuromuscular disorder and is one of the most common muscular dystrophies. There are currently few effective therapies to treat the disease, although many small-molecule approaches are being pursued. Certain histone deacetylase inhibitors (HDACi) have been shown to ameliorate DMD phenotypes in mouse and zebrafish animal models. The HDACi givinostat has shown promise for DMD in clinical trials. However, beyond a small group of HDACi, other classes of epigenetic small molecules have not been broadly and systematically studied for their benefits for DMD. We used an established animal model for DMD, the zebrafish dmd mutant strain sapje. A commercially available library of epigenetic small molecules was used to treat embryonic-larval stages of dmd mutant zebrafish. We used a quantitative muscle birefringence assay in order to assess and compare the effects of small-molecule treatments on dmd mutant zebrafish skeletal muscle structure. We performed a novel chemical-combination screen of a library of epigenetic compounds using the zebrafish dmd model. We identified candidate pools of epigenetic compounds that improve skeletal muscle structure in dmd mutant zebrafish. We then identified a specific combination of two HDACi compounds, oxamflatin and salermide, that ameliorated dmd mutant zebrafish skeletal muscle degeneration. We validated the effects of oxamflatin and salermide on dmd mutant zebrafish in an independent laboratory. Furthermore, we showed that the combination of oxamflatin and salermide caused increased levels of histone H4 acetylation in zebrafish larvae. Our results provide novel, effective methods for performing a combination of small-molecule screen in zebrafish. Our results also add to the growing evidence that epigenetic small molecules may be promising candidates for treating DMD.

中文翻译：

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斑马鱼中的一种新型化学组合筛选确定了治疗杜氏肌营养不良症的表观遗传小分子候选物

杜氏肌营养不良症 (DMD) 是一种严重的神经肌肉疾病，是最常见的肌营养不良症之一。尽管正在寻求许多小分子方法，但目前几乎没有治疗该疾病的有效疗法。某些组蛋白去乙酰化酶抑制剂 (HDACi) 已被证明可以改善小鼠和斑马鱼动物模型中的 DMD 表型。HDACi givinostat 已在临床试验中显示出 DMD 的前景。然而，除了一小组 HDACi 之外，其他类别的表观遗传小分子尚未广泛和系统地研究其对 DMD 的益处。我们使用已建立的 DMD 动物模型，即斑马鱼 dmd 突变株 sapje。商业上可获得的表观遗传小分子文库用于治疗 dmd 突变斑马鱼的胚胎幼虫阶段。我们使用定量肌肉双折射测定来评估和比较小分子治疗对 dmd 突变斑马鱼骨骼肌结构的影响。我们使用斑马鱼 dmd 模型对表观遗传化合物库进行了新的化学组合筛选。我们确定了可改善 dmd 突变斑马鱼骨骼肌结构的表观遗传化合物候选池。然后，我们确定了两种 HDACi 化合物、oxamflatin 和 salermide 的特定组合，可改善 dmd 突变体斑马鱼骨骼肌退化。我们在独立实验室中验证了oxamflatin 和 salermide 对 dmd 突变斑马鱼的影响。此外，我们表明，oxamflatin 和 Salermide 的组合导致斑马鱼幼虫组蛋白 H4 乙酰化水平增加。我们的结果提供了新颖的、在斑马鱼中进行小分子筛选组合的有效方法。我们的结果还增加了越来越多的证据表明表观遗传小分子可能是治疗 DMD 的有希望的候选者。