Frequency-dependent drug screening using optogenetic stimulation of human iPSC-derived cardiomyocytes

Sci Rep. 2017 Aug 29;7(1):9629. doi: 10.1038/s41598-017-09760-7.

Abstract

Side effects on cardiac ion channels are one major reason for new drugs to fail during preclinical evaluation. Herein we propose a simple optogenetic screening tool measuring extracellular field potentials (FP) from paced cardiomyocytes to identify drug effects over the whole physiological heart range, which is essential given the rate-dependency of ion channel function and drug action. Human induced pluripotent stem cell-derived cardiomyocytes were transduced with an adeno-associated virus to express Channelrhodopsin2 and plated on micro-electrode arrays. Global pulsed illumination (470 nm, 1 ms, 0.9 mW/mm2) was applied at frequencies from 1 to 2.5 Hz, which evoked FP simultaneously in all cardiomyocytes. This synchronized activation allowed averaging of FP from all electrodes resulting in one robust FP signal for analysis. Field potential duration (FPD) was ~25% shorter at 2.5 Hz compared to 1 Hz. Inhibition of hERG channels prolonged FPD only at low heart rates whereas Ca2+ channel block shortened FPD at all heart rates. Optogenetic pacing also allowed analysis of the maximum downstroke velocity of the FP to detect drug effects on Na+ channel availability. In principle, the presented method is well scalable for high content cardiac toxicity screening or personalized medicine for inherited cardiac channelopathies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Channelrhodopsins / analysis
  • Channelrhodopsins / genetics
  • Dependovirus / genetics
  • Drug Evaluation, Preclinical / methods*
  • Genes, Reporter
  • Genetic Vectors
  • Humans
  • Induced Pluripotent Stem Cells / drug effects*
  • Myocytes, Cardiac / drug effects*
  • Optogenetics / methods*
  • Transduction, Genetic

Substances

  • Channelrhodopsins