Drug Discovery Service

Drug Discovery Service: Cardiotoxicity Testing with Human iPSC-derived Cardiomyocytes

Using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is an accurate and useful method for predicting the proarrhythmic potential of drug candidates. This method is listed in the ICH Guidelines (S7B/E14) Q&A as a follow-up study. FUJIFILM has been dedicated to developing an assay that can generate high-quality data in a 96-well format with excellent reliability. Consequently, FUJIFILM offers a rapid and precise assay service for the evaluation of proarrhythmic potential of candidate compounds in the field of drug discovery.

iCell® Cardiomyocytes2

iCell® Cardiomyocytes2 are high-purity cardiomyocytes derived from human-induced pluripotent stem cells (hiPSCs) produced by FUJIFILM Cellular Dynamics, Inc. with proprietary differentiation technologies. They recapitulate functions of native human cardiomyocytes. With over 800 peer-reviewed publications, iCell® Cardiomyocytes2 are a valuable tool for pharmacology, safety pharmacology and basic research across multiple assay platforms. Atrial, nodal, and ventricular cardiomyocytes with spontaneous electrical activity possess typical biochemical, electrophy​siological​,​ and mechanical properties of native cardiomyocytes, as well as higher expression levels of critical genes, including ion-channels, as compared to hiPSC-derived cardiomyocytes from other sources.

 

Basic Property as Cardiomyocytes

  • High purity of cTNT positive cells1
  • Sarcomere structure2
  • Cardiac action potential2
  • Expression of major cardiac ion channels (NaV1.5, CaV1.2, hERG) comparable to human heart2

 

High-Quality Extracellular Field Potential to Assess Proarrhythmic Potential

The extracellular field potential of hiPSC-cardiomyocytes is a validated biomarker for assessing the proarrhythmic potential of drugs. It can be obtained using a multielectrode array (MEA) system3, 4. iCell® Cardiomyocytes is one of the cells used for international validation studies.

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    Robust field potential recordings with iCell® Cardiomyocytes.
  • 95184_img15R.png
    Extracellular field potentials indicating arrhythmic potential
    such as early afterdepolarization (EAD)
    are detected by specific criteria.

Identifying Proarrhythmic Potential

We established a robust protocol to obtain high quality extracellular field potential from iCell Cardiomyocytes2 by using MEA system (Maestro Pro, Axion Biosystems). Our proprietary cell culture protocol ensures optimal electrode coverage to enable the acquisition of high-quality extracellular field potential data that satisfies the analysis criteria for all wells. We can provide potential of test compounds for QT prolongation and proarrhythmia in higher throughput (up to 96well; e.g. 22 compounds at 4 different doses per plate at N=4 each dose).

Identify the electrode that provides highest quality extracellular field potential

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  • One electrode is chosen from each well that meets specific criteria for analysis to control quality of the data.

    • Signal amplitude (1st peak, 2nd peak)
    • Waveform shape
      (from depolarization to repolarization)

 

Evaluate dose-dependent FPDc prolongation and arrhythmia

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Prolongation of FPDc and occurrence of arrhythmia are evaluated for each well.

Specific criteria is set for peak detection and identification of arrhythmia to minimize human-oriented variation.

 

Visualize the proarrythmic potential of the test compounds

  • E-4031 (hERG inhibitor)
    95184_img18.png
  • Aspirin (cyclooxygenase inhibitor)
    95184_img19.png

Predictivity and Robustness

Predictivity

Results of CiPA compounds5

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The assay predicts potential risk of proarrhythmia by FPDc prolongation and occurrence of EAD in the system.

The obtained results were consistent with previously reported validation study by CiPA.

 

Robustness

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Prolongation and shortening of FPDc, as well as proarrhythmic effects, were accurately detected.
Results are consistent with previous reports, demonstrating assay robustness and repeatability.

References

  1. Ma et al., Am J Physiol Heart Circ Physiol 30 (2011) H2006-H2017
  2. Kodama et al., J Pharmacol. Sci. 140 (2019) 325-330<
  3. Blinova et al., Cell Reports 24 (2018) 3582-3592
  4. Ando et al., Journal of Pharmacological and Toxicological Methods 84 (2017) 111-127
  5. Blinova et al., Cell Reports 24 (2018) 3582-3592
  6. Kitaguchi et al., J Pharmacological and Toxicological Methods 78 (2016) 93-102
  7. Nozaki et al., Regulatory Toxicology and Pharmacology 77 (2016) 75-86

Service Flow

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MEA system (Axion BioSystems, Inc.)

  • Maestro Pro and Cytoview MEA™ plate

Cells and regents (FUJIFILM Cellular Dynamics, Inc.)

  • Cells: iCell® Cardiomyocytes2
  • Media: iCell® CM Plating Medium, iCell® CM Maintainance Medium

End Point

  • FPD/FPDc
  • Proarrhythmic Potential
    (Occurrence of EAD, Cardiac Arrest, Abnormal)
  • Beat Rate

For research use or further manufacturing use only. Not for use in diagnostic procedures.

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+49-2131-311-271

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