Frequency-Dependent Multi-Well Cardiotoxicity Screening Enabled by Optogenetic Stimulation

Autor: Daniela Malan, Heribert Bohlen, Leo Doerr, Benjamin Wolters, Susanne Rehnelt, Matthias Beckler, Philipp Sasse, Ralf Kettenhofen, Tobias Bruegmann, Krisztina Juhasz
Rok vydání: 2017
Předmět:
0301 basic medicine
Action Potentials
cardiomyocytes
Ion Channels
lcsh:Chemistry
heart rate
Medicine
Myocytes
Cardiac
Induced pluripotent stem cell
lcsh:QH301-705.5
Spectroscopy
media_common
Cardiac electrophysiology
General Medicine
ddc
Computer Science Applications
Pre-clinical development
field potential
cardiotoxicity screening
Drug
optogenetics
long QT syndrome
induced pluripotent stem cells
media_common.quotation_subject
Induced Pluripotent Stem Cells
Optogenetics
Article
Catalysis
Cell Line
Inorganic Chemistry
03 medical and health sciences
Membrane Transport Modulators
High-Throughput Screening Assays
Toxicity Tests
Humans
Physical and Theoretical Chemistry
Molecular Biology
Ion channel
Cardiotoxicity
business.industry
Organic Chemistry
Correction
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
business
Neuroscience
Zdroj: International Journal of Molecular Sciences
International Journal of Molecular Sciences; Volume 18; Issue 12; Pages: 2634
International Journal of Molecular Sciences, Vol 18, Iss 12, p 2634 (2017)
ISSN: 1422-0067
Popis: Side effects on cardiac ion channels causing lethal arrhythmias are one major reason for drug withdrawals from the market. Field potential (FP) recording from cardiomyocytes, is a well-suited tool to assess such cardiotoxic effects of drug candidates in preclinical drug development, but it is currently limited to the spontaneous beating of the cardiomyocytes and manual analysis. Herein, we present a novel optogenetic cardiotoxicity screening system suited for the parallel automated frequency-dependent analysis of drug effects on FP recorded from human pluripotent stem cell-derived cardiomyocytes. For the expression of the light-sensitive cation channel Channelrhodopsin-2, we optimised protocols using virus transduction or transient mRNA transfection. Optical stimulation was performed with a new light-emitting diode lid for a 96-well FP recording system. This enabled reliable pacing at physiologically relevant heart rates and robust recording of FP. Thereby we detected rate-dependent effects of drugs on Na+, Ca2+ and K+ channel function indicated by FP prolongation, FP shortening and the slowing of the FP downstroke component, as well as generation of afterdepolarisations. Taken together, we present a scalable approach for preclinical frequency-dependent screening of drug effects on cardiac electrophysiology. Importantly, we show that the recording and analysis can be fully automated and the technology is readily available using commercial products.
Databáze: OpenAIRE
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