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激活CRISPR可以扭转由TTN切断变体引起的哈普洛因不足和功能缺陷

Shahnaz Ghahremani¹, Aditya Kanwal¹, Anthony Pettinato²

¹The Jackson Laboratory for Genomic Medicine, Farmington, CT (S.G., A.K., N.L., K.T., Y.Z., H.T., C.-L.W., J.T.H.).

|January 18, 2024

概括

在TTN变异引起的扩张性心肌病模型中,TTN基因的CRISPR激活成功恢复了心肌功能. 这种方法为大量患有这种疾病的患者提供了潜在的治疗策略.

科学领域:

心血管遗传学
基因治疗
分子心脏病学

背景情况:

在TTN基因 (TTNtvs) 中的缩减变异是扩张性心肌病的主要遗传原因,
TTNtvs导致功能性蛋白减少,影响瘤完整性和心脏功能,治疗选择有限.
开发疗法的挑战包括TTN基因的大小,TTNtvs的多样性以及对其致病性的不完全理解.

研究的目的:

适应CRISPR激活技术用于TTNtv致病性的功能性查询.
使用CRISPR激活开发TNTvs引起的扩张性心肌病的潜在治疗策略.
在患者心脏模型中研究TTN基因激活的分子后果和治疗效果.

主要方法:

使用dCas9-VPR激活人类心肌细胞和来自带有TTNtv的诱导多能干细胞的3D心脏微组织.
使用定制的TTN报告测试和凝电泳来量化TTN蛋白质水平和异型.
进行RNA测序和心肌细胞表观遗传分析以确定分子效应,并指定心肌细胞特异性TTN激活的调节元素.

主要成果:

对TTN的CRISPR激活成功地挽救了TTNtvs引起的TTN蛋白缺陷.
增加的TTN蛋白质水平使瘤含量和心脏收缩功能正常化,即使TTN蛋白质的含量增加.
克里斯普尔激活对TTN转录以及参与肌纤维组合和瘤结构的转录进行了上调.

结论:

对TTN的CRISPR激活有效地扭转了与TTNtvs相关的功能缺陷,支持了作为一个关键机制的哈普洛因不足.
这项研究表明TTN CRISPR激活作为治疗广泛的TTNtv相关扩张性心肌病的潜在策略.
这些发现为开发针对心力衰竭TTN变异的基因疗法铺平了道路.

关键词:

心肌病,扩大聚类定期间隔的简短的平行体重复连接肌细胞,心脏

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