PDF(Pubmed)
Abstract：
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in combination with CRISPR/Cas9 genome editing provide unparalleled opportunities to study cardiac biology and disease. However, sarcomeres, the fundamental units of myocyte contraction, are immature and nonlinear in hiPSC-CMs, which technically challenge accurate functional interrogation of contractile parameters in beating cells. Furthermore, existing analysis methods are relatively low-throughput, indirectly assess contractility, or only assess well-aligned sarcomeres found in mature cardiac tissues.
We aimed to develop an analysis platform that directly, rapidly, and automatically tracks sarcomeres in beating cardiomyocytes. The platform should assess sarcomere content, contraction and relaxation parameters, and beat rate.
We developed SarcTrack, a MatLab software that monitors fluorescently tagged sarcomeres in hiPSC-CMs. The algorithm determines sarcomere content, sarcomere length, and returns rates of sarcomere contraction and relaxation. By rapid measurement of hundreds of sarcomeres in each hiPSC-CM, SarcTrack provides large data sets for robust statistical analyses of multiple contractile parameters. We validated SarcTrack by analyzing drug-treated hiPSC-CMs, confirming the contractility effects of compounds that directly activate (CK-1827452) or inhibit (MYK-461) myosin molecules or indirectly alter contractility (verapamil and propranolol). SarcTrack analysis of hiPSC-CMs carrying a heterozygous truncation variant in the myosin-binding protein C ( MYBPC3) gene, which causes hypertrophic cardiomyopathy, recapitulated seminal disease phenotypes including cardiac hypercontractility and diminished relaxation, abnormalities that normalized with MYK-461 treatment.
SarcTrack provides a direct and efficient method to quantitatively assess sarcomere function. By improving existing contractility analysis methods and overcoming technical challenges associated with functional evaluation of hiPSC-CMs, SarcTrack enhances translational prospects for sarcomere-regulating therapeutics and accelerates interrogation of human cardiac genetic variants.
We aimed to develop an analysis platform that directly, rapidly, and automatically tracks sarcomeres in beating cardiomyocytes. The platform should assess sarcomere content, contraction and relaxation parameters, and beat rate.
We developed SarcTrack, a MatLab software that monitors fluorescently tagged sarcomeres in hiPSC-CMs. The algorithm determines sarcomere content, sarcomere length, and returns rates of sarcomere contraction and relaxation. By rapid measurement of hundreds of sarcomeres in each hiPSC-CM, SarcTrack provides large data sets for robust statistical analyses of multiple contractile parameters. We validated SarcTrack by analyzing drug-treated hiPSC-CMs, confirming the contractility effects of compounds that directly activate (CK-1827452) or inhibit (MYK-461) myosin molecules or indirectly alter contractility (verapamil and propranolol). SarcTrack analysis of hiPSC-CMs carrying a heterozygous truncation variant in the myosin-binding protein C ( MYBPC3) gene, which causes hypertrophic cardiomyopathy, recapitulated seminal disease phenotypes including cardiac hypercontractility and diminished relaxation, abnormalities that normalized with MYK-461 treatment.
SarcTrack provides a direct and efficient method to quantitatively assess sarcomere function. By improving existing contractility analysis methods and overcoming technical challenges associated with functional evaluation of hiPSC-CMs, SarcTrack enhances translational prospects for sarcomere-regulating therapeutics and accelerates interrogation of human cardiac genetic variants.
摘要：
人类诱导多能干细胞衍生的心肌细胞（hiPSC-CM）与CRISPR/Cas9基因组编辑相结合，为研究心脏生物学和疾病提供了无与伦比的机会。然而,肉瘤,肌细胞收缩的基本单位，在HiPSC-CM中是不成熟和非线性的，这在技术上挑战了对搏动细胞收缩参数的准确功能询问。此外，现有的分析方法吞吐量相对较低，间接评估收缩性，或仅评估在成熟的心脏组织中发现的对齐良好的肉瘤。
我们的目标是开发一个分析平台，迅速,并自动跟踪搏动心肌细胞中的肉瘤。该平台应该评估sarcomere内容，收缩和松弛参数，和节拍率。
我们开发了SarcTrack,aMatLab软件，用于监控hiPSC-CM中荧光标记的sarcome。该算法确定sarcome含量，肌节长度,以及肌节收缩和松弛的回报率。通过快速测量每个hiPSC-CM中的数百个肉瘤，SarcTrack为多个收缩参数的强大统计分析提供了大量数据集。我们通过分析药物治疗的hiPSC-CM来验证SarcTrack，确认直接激活（CK-1827452）或抑制（MYK-461）肌球蛋白分子或间接改变收缩性（维拉帕米和普萘洛尔）的化合物的收缩作用。在肌球蛋白结合蛋白C（MYBPC3）基因中携带杂合截短变体的hiPSC-CM的SarcTrack分析，导致肥厚型心肌病，概述了精液疾病的表型，包括心脏收缩过度和松弛减少，用MYK-461治疗恢复正常的异常。
SarcTrack提供了一种直接有效的方法来定量评估肌节功能。通过改进现有的收缩性分析方法并克服与hiPSC-CM功能评估相关的技术挑战，SarcTrack增强了肌节调节疗法的翻译前景，并加速了人类心脏遗传变异的询问。
我们的目标是开发一个分析平台，
我们开发了SarcTrack,
SarcTrack提供了一种直接有效的方法来定量评估肌节功能。
