In Vitro Pharmacology >> Ion Channels >> Cardiotoxicity

Human Cardiomyocytes Cardiotoxicity Services

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Multi-prong Assessment of Cardiac Liability

Cardiotoxicity is a major cause of drug development failure and withdrawals. hERG screening alone cannot reliably detect potential cardiac adverse side effects. The Cardiac Safety Research Consortium (CSRC) recently proposed the Comprehensive in vitro Proarrhythmia Assay (CiPA), which would incorporate in vitro cardiomyocyte studies as a component of cardiac safety assessment. In addition to our CardiacProfiler™ services, our services using human cardiomyocytes fit into this emerging paradigm.

We have employed a combination of assay platforms along with Cytiva™ Plus human stem cell derived cardiomyocytes from GE Healthcare to address this need for more biologically relevant and predictive cardiac liability models. Eurofins offers complimentary approaches, including High Content Analsysis (HCA), Multiple Electrode Array (MEA) and manual patch clamp assays, to reliably detect and characterize cardiac safety risks.

Using a population of beating cardiomyocytes, MEA provides ECG-like field potentials to detect proarrhythmic events, alterations in beat rate and other adverse effects. Changes in field potential duration is related to alterations of the QT interval, which in a clinical setting may initiate a potentially fatal Torsades de Pointes (TdP) arrhythmia.

Manual patch clamp studies provide a detailed assessment of action potential duration on individual cardiomyocytes. Specifically, ventricular-type cells within the general cardiomyocyte population can be tested. This is important as the ventricular-type cardiomyocytes are relevant for ventricular dysfunction, a major form of cardiotoxicity.

Imaging techniques using cardiomyocytes measure cell health parameters resulting from structural damage upon prolonged compound treatment, toxic effects that may not be otherwise identified by electrophysiological methods. Multiple parameters, including cell number, intracellular calcium flux, mitochondrial membrane potential and membrane permeability can be measured on live cells to determine changes in normal cellular functions. Alternatively, fixed cells are used to reveal appearance of peri-nuclear BNP expression, α-actinin disruption, troponin I integrity and reduction in cell number to further demonstrate toxic effects.

Advantages of Cardiotoxicity Assessment with Eurofins:

  • Biologically relevant, highly characterized and reproducible cardiac cell model:
    • Differentiated human stem cells provide a reliable and reproducible source of cardiomyocytes
    • Highly pure cell population with cardiac marker expression (α-actinin, troponin I)
    • Large fraction of ventricular subtype cells to test ventricular dysfunction - the predominant form of cardiotoxicity
    • Functionally verified by patch clamp for both action potential and individual ion channel currents
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Figure 1. Changes to cardiomyocytes field potential duration (FPD) resulting from treatment with known ion channel and GPCR modulators as detected by MEA.

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Figure 2. Effect of quinidine on ventricular-type cardiomyocyte action potential by manual patch clamp.

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Figure 3. Functional and structural modifications of cardiomyocyte by sunitinib using HCA.

Cardiomyocytes were either treated with vehicle (top panels) or with sunitinib (bottom panels) and then monitored for changes in calcium mobilization (left panels), mitochondrial membrane potential (center panels) and membrane permeability (right panels).