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Use of human induced pluripotent stem cell–derived cardiomyocytes to assess drug cardiotoxicity

Abstract

Cardiotoxicity has historically been a major cause of drug removal from the pharmaceutical market. Several chemotherapeutic compounds have been noted for their propensities to induce dangerous cardiac-specific side effects such as arrhythmias or cardiomyocyte apoptosis. However, improved preclinical screening methodologies have enabled cardiotoxic compounds to be identified earlier in the drug development pipeline. Human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) can be used to screen for drug-induced alterations in cardiac cellular contractility, electrophysiology, and viability. We previously established a novel ‘cardiac safety index’ (CSI) as a metric that can evaluate potential cardiotoxic drugs via high-throughput screening of hiPSC-CMs. This metric quantitatively examines drug-induced alterations in CM function, using several in vitro readouts, and normalizes the resulting toxicity values to the in vivo maximum drug blood plasma concentration seen in preclinical or clinical pharmacokinetic models. In this ~1-month-long protocol, we describe how to differentiate hiPSCs into hiPSC-CMs and subsequently implement contractility and cytotoxicity assays that can evaluate drug-induced cardiotoxicity in hiPSC-CMs. We also describe how to carry out the calculations needed to generate the CSI metric from these quantitative toxicity measurements.

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Acknowledgements

We acknowledge the Stanford High-Throughput Bioscience Center for assistance with high-throughput imaging and plate reader assays. We acknowledge support from an American Heart Association (AHA) Predoctoral Fellowship (13PRE15770000) and an NSF Graduate Research Fellowship (DGE-114747) (A.S.); Burroughs Wellcome Foundation Innovation grant 1015009, AHA grant 17MERIT3361009, and National Institutes of Health (NIH) grants R01 HL132875, R01 HL130020, R01 HL128170, R01 HL123968, and R24 HL117756 (J.C.W.); and NIH grants NIH R01 HL130840, R01 HL128072, and R01 HL132225 (M.M.).

Author information

A.S. designed, supervised, and performed experiments, and wrote the manuscript with help from all authors. W.L.M. performed high-throughput experiments related to the contractility analysis. R.S. developed analysis platforms for quantitative assessment of hiPSC-CM contractility. T.K. performed experiments related to hiPSC-CM cytotoxicity analysis. P.W.B. provided expertise related to hiPSC-CM platform development. J.C.d.Á. provided mathematical expertise related to hiPSC-CM contractility analysis. M.M. provided advice and proofread the manuscript. J.C.W. provided advice, proofread the manuscript, and provided financial support for experiments.

Correspondence to Joseph C. Wu.

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Competing interests

M.M. holds equity in and is on the scientific advisory board for Vala Sciences, a company offering high-content screening services and instrumentation for measuring the electrical and contractile physiology of CMs. J.C.W. is a cofounder of and scientific advisory board member for Khoris, a company using hiPSCs to accelerate drug discovery. The remaining authors declare no competing interests.

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Key references using this protocol

Sharma, A. et al. Sci. Transl. Med. 9, eaaf2584 (2017): https://doi.org/10.1126/scitranslmed.aaf2584

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Further reading

Fig. 1: Workflow for generating the cardiac safety index for drugs of interest using high-throughput contractility and cytotoxicity analysis of hiPSC-CMs.
Fig. 2: Procedure for chemically defined differentiation of hiPSCs into hiPSC-CMs and downstream replating.
Fig. 3: Setup for drug-induced cytotoxicity and contractility assessment of hiPSC-CMs.
Fig. 4: Equations used to obtain the CSI.
Fig. 5: Mathematical example of CSI calculations for cardiotoxic tyrosine kinase inhibitor sorafenib.

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