Allele-specific RNA silencing has been shown to be an effective therapeutic treatment in a number of diseases, including neurodegenerative disorders. Studies of allele-specific silencing in hypertrophic cardiomyopathy (HCM) to date have focused on mouse models of disease. We here examine allele-specific silencing in a human-cell model of HCM. We investigate two methods of silencing, short hairpin RNA (shRNA) and antisense oligonucleotide (ASO) silencing, using a human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) model. We used cellular micropatterning devices with traction force microscopy and automated video analysis to examine each strategy's effects on contractile defects underlying disease. We find that shRNA silencing ameliorates contractile phenotypes of disease, reducing disease-associated increases in cardiomyocyte velocity, force, and power. We find that ASO silencing, while better able to target and knockdown a specific disease-associated allele, showed more modest improvements in contractile phenotypes. These findings are the first exploration of allele-specific silencing in a human HCM model and provide a foundation for further exploration of silencing as a therapeutic treatment for MYH7-mutation-associated cardiomyopathy.

中文翻译：

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MYH7 的沉默改善了人类 iPSC 心肌细胞的疾病表型。

等位基因特异性 RNA 沉默已被证明是许多疾病的有效治疗方法，包括神经退行性疾病。迄今为止，肥厚型心肌病 (HCM) 中等位基因特异性沉默的研究主要集中在小鼠疾病模型上。我们在这里检查 HCM 人类细胞模型中的等位基因特异性沉默。我们使用人类诱导多能干细胞衍生的心肌细胞 (hiPSC-CM) 模型研究了两种沉默方法，即短发夹 RNA (shRNA) 和反义寡核苷酸 (ASO) 沉默。我们使用带有牵引力显微镜和自动视频分析的细胞微图案设备来检查每种策略对潜在疾病的收缩缺陷的影响。我们发现 shRNA 沉默改善了疾病的收缩表型，减少与疾病相关的心肌细胞速度、力量和力量的增加。我们发现 ASO 沉默虽然能够更好地靶向和击倒特定的疾病相关等位基因，但在收缩表型方面表现出更适度的改善。这些发现是人类 HCM 模型中等位基因特异性沉默的首次探索，并为进一步探索沉默作为 MYH7 突变相关心肌病的治疗方法提供了基础。