High Throughput Measurement of Ca²⁺ Dynamics for Drug Risk Assessment in Human Stem Cell-derived Cardiomyocytes by Kinetic Image Cytometry

Overview

Journal J Pharmacol Toxicol Methods

Publisher Elsevier

Specialties Pharmacology
Toxicology

Date 2012 Aug 29

PMID 22926323

Citations 51

Authors

Fabio Cerignoli

David Charlot

Ross Whittaker

Randy Ingermanson

Piyush Gehalot

Alex Savchenko

David J Gallacher

Rob Towart

Jeffrey H Price

Patrick M McDonough

Mark Mercola

Affiliations

Soon will be listed here.

Abstract

Current methods to measure physiological properties of cardiomyocytes and predict fatal arrhythmias that can cause sudden death, such as Torsade de Pointes, lack either the automation and throughput needed for early-stage drug discovery and/or have poor predictive value. To increase throughput and predictive power of in vitro assays, we developed kinetic imaging cytometry (KIC) for automated cell-by-cell analyses via intracellular fluorescence Ca²⁺ indicators. The KIC instrument simultaneously records and analyzes intracellular calcium concentration [Ca²⁺](i) at 30-ms resolution from hundreds of individual cells/well of 96-well plates in seconds, providing kinetic details not previously possible with well averaging technologies such as plate readers. Analyses of human embryonic stem cell and induced pluripotent stem cell-derived cardiomyocytes revealed effects of known cardiotoxic and arrhythmogenic drugs on kinetic parameters of Ca²⁺ dynamics, suggesting that KIC will aid in the assessment of cardiotoxic risk and in the elucidation of pathogenic mechanisms of heart disease associated with drugs treatment and/or genetic background.

Citing Articles

Modulation of Ca oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis.

Sasaki T, Hisada S, Kanki H, Nunomura K, Lin B, Nishiyama K Sci Rep. 2024; 14(1):27667.

PMID: 39532929 PMC: 11557898. DOI: 10.1038/s41598-024-77882-w.

Characterizing PFAS hazards and risks: a human population-based in vitro cardiotoxicity assessment strategy.

Ford L, Lin H, Zhou Y, Wright F, Gombar V, Sedykh A Hum Genomics. 2024; 18(1):92.

PMID: 39218963 PMC: 11368000. DOI: 10.1186/s40246-024-00665-x.

BeatProfiler: Multimodal In Vitro Analysis of Cardiac Function Enables Machine Learning Classification of Diseases and Drugs.

Kim Y, Wang K, Lock R, Nash T, Fleischer S, Wang B IEEE Open J Eng Med Biol. 2024; 5:238-249.

PMID: 38606403 PMC: 11008807. DOI: 10.1109/OJEMB.2024.3377461.

OptoDyCE-plate as an affordable high throughput imager for all optical cardiac electrophysiology.

Heinson Y, Han J, Entcheva E J Mol Cell Cardiol Plus. 2023; 6.

PMID: 38130942 PMC: 10735237. DOI: 10.1016/j.jmccpl.2023.100054.

OptoDyCE-plate as an affordable high throughput imager for all optical cardiac electrophysiology.

Heinson Y, Han J, Entcheva E bioRxiv. 2023; .

PMID: 37693544 PMC: 10491208. DOI: 10.1101/2023.08.29.555447.

References

Klein I, Danzi S . Thyroid disease and the heart. Circulation. 2007; 116(15):1725-35. DOI: 10.1161/CIRCULATIONAHA.106.678326. View

Fernandez-Velasco M, Rueda A, Rizzi N, Benitah J, Colombi B, Napolitano C . Increased Ca2+ sensitivity of the ryanodine receptor mutant RyR2R4496C underlies catecholaminergic polymorphic ventricular tachycardia. Circ Res. 2008; 104(2):201-9, 12p following 209. PMC: 2796688. DOI: 10.1161/CIRCRESAHA.108.177493. View

Komukai K, O-Uchi J, Morimoto S, Kawai M, Hongo K, Yoshimura M . Role of Ca(2+)/calmodulin-dependent protein kinase II in the regulation of the cardiac L-type Ca(2+) current during endothelin-1 stimulation. Am J Physiol Heart Circ Physiol. 2010; 298(6):H1902-7. DOI: 10.1152/ajpheart.01141.2009. View

Olson R, Gambliel H, Vestal R, Shadle S, Charlier Jr H, Cusack B . Doxorubicin cardiac dysfunction: effects on calcium regulatory proteins, sarcoplasmic reticulum, and triiodothyronine. Cardiovasc Toxicol. 2005; 5(3):269-83. DOI: 10.1385/ct:5:3:269. View

Monteith G, Bird G . Techniques: high-throughput measurement of intracellular Ca(2+) -- back to basics. Trends Pharmacol Sci. 2005; 26(4):218-23. DOI: 10.1016/j.tips.2005.02.002. View

Braam S, Tertoolen L, van de Stolpe A, Meyer T, Passier R, Mummery C . Prediction of drug-induced cardiotoxicity using human embryonic stem cell-derived cardiomyocytes. Stem Cell Res. 2009; 4(2):107-16. DOI: 10.1016/j.scr.2009.11.004. View

Choi B, Burton F, Salama G . Cytosolic Ca2+ triggers early afterdepolarizations and Torsade de Pointes in rabbit hearts with type 2 long QT syndrome. J Physiol. 2002; 543(Pt 2):615-31. PMC: 2290501. DOI: 10.1113/jphysiol.2002.024570. View

Mohammad S, Zhou Z, Gong Q, January C . Blockage of the HERG human cardiac K+ channel by the gastrointestinal prokinetic agent cisapride. Am J Physiol. 1997; 273(5):H2534-8. DOI: 10.1152/ajpheart.1997.273.5.H2534. View

Bode G, Olejniczak K . ICH topic: the draft ICH S7B step 2: note for guidance on safety pharmacology studies for human pharmaceuticals. Fundam Clin Pharmacol. 2002; 16(2):105-18. DOI: 10.1046/j.1472-8206.2002.00079.x. View

10.

Bootman M, Fearnley C, Smyrnias I, MacDonald F, Roderick H . An update on nuclear calcium signalling. J Cell Sci. 2009; 122(Pt 14):2337-50. DOI: 10.1242/jcs.028100. View

11.

Hondeghem L . Use and abuse of QT and TRIaD in cardiac safety research: importance of study design and conduct. Eur J Pharmacol. 2008; 584(1):1-9. DOI: 10.1016/j.ejphar.2008.01.016. View

12.

Fermini B, Fossa A . The impact of drug-induced QT interval prolongation on drug discovery and development. Nat Rev Drug Discov. 2003; 2(6):439-47. DOI: 10.1038/nrd1108. View

13.

Guth B . Preclinical cardiovascular risk assessment in modern drug development. Toxicol Sci. 2007; 97(1):4-20. DOI: 10.1093/toxsci/kfm026. View

14.

Patmore L, Fraser S, Mair D, Templeton A . Effects of sparfloxacin, grepafloxacin, moxifloxacin, and ciprofloxacin on cardiac action potential duration. Eur J Pharmacol. 2000; 406(3):449-52. DOI: 10.1016/s0014-2999(00)00694-4. View

15.

Hove-Madsen L, Llach A, Molina C, Prat-Vidal C, Farre J, Roura S . The proarrhythmic antihistaminic drug terfenadine increases spontaneous calcium release in human atrial myocytes. Eur J Pharmacol. 2006; 553(1-3):215-21. DOI: 10.1016/j.ejphar.2006.09.023. View

16.

Valentin J, Hoffmann P, De Clerck F, Hammond T, Hondeghem L . Review of the predictive value of the Langendorff heart model (Screenit system) in assessing the proarrhythmic potential of drugs. J Pharmacol Toxicol Methods. 2004; 49(3):171-81. DOI: 10.1016/j.vascn.2004.03.008. View

17.

Houjou S, Iizuka K, Dobashi K, Nakazawa T . Allergic responses reduce the relaxant effect of beta-agonists but not potassium channel openers in guinea-pig isolated trachea. Eur Respir J. 1996; 9(10):2050-6. DOI: 10.1183/09031936.96.09102050. View

18.

Towart R, Linders J, Hermans A, Rohrbacher J, van der Linde H, Ercken M . Blockade of the I(Ks) potassium channel: an overlooked cardiovascular liability in drug safety screening?. J Pharmacol Toxicol Methods. 2009; 60(1):1-10. DOI: 10.1016/j.vascn.2009.04.197. View

19.

Ogawa S, BARNETT J, Sen L, Galper J, Smith T, Marsh J . Direct contact between sympathetic neurons and rat cardiac myocytes in vitro increases expression of functional calcium channels. J Clin Invest. 1992; 89(4):1085-93. PMC: 442964. DOI: 10.1172/JCI115688. View

20.

Balasubramaniam R, Chawla S, Grace A, Huang C . Caffeine-induced arrhythmias in murine hearts parallel changes in cellular Ca(2+) homeostasis. Am J Physiol Heart Circ Physiol. 2005; 289(4):H1584-93. DOI: 10.1152/ajpheart.01250.2004. View