Non-negative blind deconvolution for signal processing in a CRISPR-edited iPSC-cardiomyocyte model of dilated cardiomyopathy

FEBS Lett. 2021 Oct;595(20):2544-2557. doi: 10.1002/1873-3468.14189. Epub 2021 Sep 17.

Abstract

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 (Ca2+ ) 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.

Keywords: atomic force microscopy; cardiovascular disease; contractility; dilated cardiomyopathy; human-induced pluripotent stem cell-derived cardiomyocytes; microdomains; signal transduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems*
  • Cardiomyopathy, Dilated / genetics
  • Cardiomyopathy, Dilated / pathology*
  • Gene Editing*
  • Humans
  • Induced Pluripotent Stem Cells / pathology*
  • Models, Biological*
  • Mutation
  • Myocytes, Cardiac / pathology*
  • Troponin T / genetics

Substances

  • Troponin T