Heart Research Center, Department of Cardiology and Pneumology, University Medical Center, Goettingen University, Germany.
DZHK (German Center for Cardiovascular Research), Partner Site Goettingen, Germany.
FEBS Lett. 2021 Oct;595(20):2544-2557. doi: 10.1002/1873-3468.14189. Epub 2021 Sep 17.
We developed an integrated platform for analysis of parameterized data from human disease models. We report a non-negative blind deconvolution (NNBD) approach to quantify calcium (Ca ) handling, beating force and contractility in human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) at the single-cell level. We employed CRISPR/Cas gene editing to introduce a dilated cardiomyopathy (DCM)-causing mutation in troponin T (TnT), TnT-R141W, into wild-type control iPSCs (MUT). The NNDB-based method enabled data parametrization, fitting and analysis in wild-type controls versus isogenic MUT iPSC-CMs. Of note, Cas9-edited TnT-R141W iPSC-CMs revealed significantly reduced beating force and prolonged contractile event duration. The NNBD-based platform provides an alternative framework for improved quantitation of molecular disease phenotypes and may contribute to the development of novel diagnostic tools.
我们开发了一个用于分析人类疾病模型中参数化数据的综合平台。我们报告了一种非负盲解卷积（NNBD）方法，用于在单细胞水平上定量人诱导多能干细胞衍生的心肌细胞（iPSC-CM）中的钙（Ca ）处理、搏动力和收缩性。我们利用 CRISPR/Cas 基因编辑将肌钙蛋白 T（TnT）中的扩张型心肌病（DCM）致病突变 TnT-R141W 引入野生型对照 iPSC（MUT）。基于 NNBD 的方法能够对野生型对照与同基因 MUT iPSC-CM 进行数据参数化、拟合和分析。值得注意的是，Cas9 编辑的 TnT-R141W iPSC-CM 显示出明显降低的搏动力和延长的收缩事件持续时间。基于 NNBD 的平台提供了一种改进分子疾病表型定量的替代框架，并可能有助于开发新的诊断工具。
