Abstract
The ability to produce unlimited numbers of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) harboring disease and patient-specific gene variants creates a new paradigm for modeling congenital heart diseases (CHDs) and predicting proarrhythmic liabilities of drug candidates. However, a major roadblock to implementing hiPSC-CM technology in drug discovery is that conventional methods for monitoring action potential (AP) kinetics and arrhythmia phenotypes in vitro have been too costly or technically challenging to execute in high throughput. Herein, we describe the first large-scale, fully automated and statistically robust analysis of AP kinetics and drug-induced proarrhythmia in hiPSC-CMs. The platform combines the optical recording of a small molecule fluorescent voltage sensing probe (VoltageFluor2.1.Cl), an automated high throughput microscope and automated image analysis to rapidly generate physiological measurements of cardiomyocytes (CMs). The technique can be readily adapted on any high content imager to study hiPSC-CM physiology and predict the proarrhythmic effects of drug candidates.
Highlights
Between 1991 and 2003, 6 drugs were withdrawn from the market in the U.S because of evidence that they induced QT prolongation and the potentially fatal ventricular tachycardia (VT), Torsade de Pointes (TdP) (Waring et al, 2015)
To measure changes in membrane potential, the platform utilized VF2.1.Cl (Miller et al, 2012), which intercalates into the plasma membrane and senses changes in transmembrane potential by a novel photo-induced electron transfer (PeT) mechanism (Miller, 2016; VF2.1.Cl and related voltage sensitive probe (VSP) have been described in detail Woodford et al, 2015; Deal et al, 2016; Kulkarni et al, 2017, and can be obtained directly from EWM)
Using an optical method for high throughput detection of action potential (AP) kinetics, we developed an automated platform for assessment of proarrhythmia in normal and channelopathy patient derived hiPSC-CMs
Summary
Between 1991 and 2003, 6 drugs were withdrawn from the market in the U.S because of evidence that they induced QT prolongation and the potentially fatal ventricular tachycardia (VT), Torsade de Pointes (TdP) (Waring et al, 2015). The in vitro measurement of hERG inhibition has become essential for advancement of a drug candidate to clinical development (Waring et al, 2015). HERG inhibition is highly sensitive to a wide range of chemical substructures (Sanguinetti and Tristani-Firouzi, 2006) that might comprise clinically beneficial components of small molecule drugs. There is considerable concern that reliance on hERG inhibition alone may prevent many useful compounds from advancing toward the clinic. This concern was embodied in the 2013 Food and Drug Administration recognition of the need for a comprehensive non-clinical assay to better predict the proarrhythmic potential of new drugs (Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative) (Sager et al, 2014)
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