EFP Analyzer: A fast, accurate, and easy-to-teach program for analyzing Extracellular Field Potentials from iPSC-derived cardiomyocytes

Patel, N. P.; Shen, A.; Wada, Y.; Blair, M. A.; Mitchell, D. W.; Vanags, L. R.; Woo, S.; Ku, M. J.; Dauda, K. S.; Morris, W. A.; Yang, M.; Knollmann, B. C.; Roden, D. M.; Salem, J. E.; Glazer, A. M.; Kroncke, B. M.

doi:10.1101/2024.10.21.619481

bioRxiv · 10.1101/2024.10.21.619481

EFP Analyzer: A fast, accurate, and easy-to-teach program for analyzing Extracellular Field Potentials from iPSC-derived cardiomyocytes

Patel, N. P.·Shen, A.·Wada, Y.·Blair, M. A.·Mitchell, D. W.·Vanags, L. R.·Woo, S.·Ku, M. J.·Dauda, K. S.·Morris, W. A.·Yang, M.·Knollmann, B. C.·Roden, D. M.·Salem, J. E.·Glazer, A. M.·Kroncke, B. M.

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Abstract

RationaleInduced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are an emerging model for determining drug effects and modeling disease. Specialized devices can generate Extracellular Field Potential (EFP) measurements from these cells, analogous to the ventricular complex of the electrocardiogram. ObjectiveThe objective of this study was to develop an easy-to-use, easy-to-teach, reproducible software tool to measure EFPs. Methods-ResultsWe present the EFP-Analyzer (EFPA), a semi-automized analyzer for EFP traces, which identifies and averages beats, identifies landmarks, and calculates intervals. We evaluated the tool in an analysis of 358 EFP traces from 22 patient-derived lines. We analyzed spontaneously beating iPSC-CMs, as well as optically paced iPSC-CMs through channelrhodopsin. We developed stringent quality criteria and measured EFP intervals, including Field Potential Duration (FPD). FPD from optically paced iPSC-CMs were shorter than those of spontaneously beating iPSC-CMs (283.7.0{+/-}54.2 vs. 293.0{+/-}47.5, p: 0.32, respectively). We further analyzed the usability and data replicability of EFPA through an inter-intra observer analysis. Correlation coefficient for inter-reader tangent and threshold measurements for these FPD ranged between r: 0.93-1.00. Bland-Altman plots comparing inter observer results for spontaneously beating and paced iPSC-CMs showed 95% limits of agreement (-13.6 to 19.4ms and -13.2 to 15.3ms, respectively). The EFP-analyzer could accurately detect FPD prolongation due to drug (moxifloxacin) or pathogenic loss of function mutations (CACNA1C N639T). This program is available for download at https://github.com/kroncke-lab/EFPA. The instructions will be available at the same listed website under the README section of the Github main page. ConclusionsThe EFP-Analyzer tool is a useful tool that enables the efficient use of iPSC-CMs as a model to study drug effects and disease.

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Patel, N. P., Shen, A., Wada, Y., Blair, M. A., Mitchell, D. W., Vanags, L. R., Woo, S., Ku, M. J., Dauda, K. S., Morris, W. A., Yang, M., Knollmann, B. C., Roden, D. M., Salem, J. E., Glazer, A. M., Kroncke, B. M.. 2024-10-24. EFP Analyzer: A fast, accurate, and easy-to-teach program for analyzing Extracellular Field Potentials from iPSC-derived cardiomyocytes. https://doi.org/10.1101/2024.10.21.619481

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