Modeling susceptibility to drug-induced long QT with a panel of subject-specific induced pluripotent stem cells
Francesca Stillitano, Jens Hansen, Chi-Wing Kong, Ioannis Karakikes, Christian Funck-Brentano, Lin Geng, Stuart Scott, Stephan Reynier, Ma Wu, Yannick Valogne, Carole Desseaux, Joe-Elie Salem, Dorota Jeziorowska, Noël Zahr, Ronald Li, Ravi Iyengar, Roger J Hajjar, Jean-Sébastien Hulot, Francesca Stillitano, Jens Hansen, Chi-Wing Kong, Ioannis Karakikes, Christian Funck-Brentano, Lin Geng, Stuart Scott, Stephan Reynier, Ma Wu, Yannick Valogne, Carole Desseaux, Joe-Elie Salem, Dorota Jeziorowska, Noël Zahr, Ronald Li, Ravi Iyengar, Roger J Hajjar, Jean-Sébastien Hulot
Abstract
A large number of drugs can induce prolongation of cardiac repolarization and life-threatening cardiac arrhythmias. The prediction of this side effect is however challenging as it usually develops in some genetically predisposed individuals with normal cardiac repolarization at baseline. Here, we describe a platform based on a genetically diverse panel of induced pluripotent stem cells (iPSCs) that reproduces susceptibility to develop a cardiotoxic drug response. We generated iPSC-derived cardiomyocytes from patients presenting in vivo with extremely low or high changes in cardiac repolarization in response to a pharmacological challenge with sotalol. In vitro, the responses to sotalol were highly variable but strongly correlated to the inter-individual differences observed in vivo. Transcriptomic profiling identified dysregulation of genes (DLG2, KCNE4, PTRF, HTR2C, CAMKV) involved in downstream regulation of cardiac repolarization machinery as underlying high sensitivity to sotalol. Our findings offer novel insights for the development of iPSC-based screening assays for testing individual drug reactions.
Trial registration: ClinicalTrials.gov NCT01338441.
Keywords: arrhythmia; cardiotoxicity; human; human biology; induced pluripotent stem cells; medicine.
Conflict of interest statement
SR: Current or former employee of the Cellectis Company. MW: Current or former employee of the Cellectis Company. YV: Current or former employee of the Cellectis Company. CD: Current or former employee of the Cellectis Company. The other authors declare that no competing interests exist.
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References
- Behr ER, Roden D. Drug-induced arrhythmia: pharmacogenomic prescribing? European Heart Journal. 2013;34:89–95. doi: 10.1093/eurheartj/ehs351.
- Berger SI, Ma'ayan A, Iyengar R. Systems pharmacology of arrhythmias. Science Signaling. 2010;3:ra30. doi: 10.1126/scisignal.2000723.
- Budnitz DS, Pollock DA, Weidenbach KN, Mendelsohn AB, Schroeder TJ, Annest JL. National surveillance of emergency department visits for outpatient adverse drug events. JAMA. 2006;296:1858–1866. doi: 10.1001/jama.296.15.1858.
- Burridge PW, Matsa E, Shukla P, Lin ZC, Churko JM, Ebert AD, Lan F, Diecke S, Huber B, Mordwinkin NM, Plews JR, Abilez OJ, Cui B, Gold JD, Wu JC. Chemically defined generation of human cardiomyocytes. Nature Methods. 2014;11:855–860. doi: 10.1038/nmeth.2999.
- Cerrone M, Priori SG. Genetics of sudden death: focus on inherited channelopathies. European Heart Journal. 2011;32:2109–2118. doi: 10.1093/eurheartj/ehr082.
- Chen EY, Tan CM, Kou Y, Duan Q, Wang Z, Meirelles GV, Clark NR, Ma'ayan A. Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinformatics. 2013;14:128. doi: 10.1186/1471-2105-14-128.
- Chen EY, Xu H, Gordonov S, Lim MP, Perkins MH, Ma'ayan A. Expression2Kinases: mRNA profiling linked to multiple upstream regulatory layers. Bioinformatics. 2012;28:105–111. doi: 10.1093/bioinformatics/btr625.
- Cirulli ET, Goldstein DB. Uncovering the roles of rare variants in common disease through whole-genome sequencing. Nature Reviews Genetics. 2010;11:415–425. doi: 10.1038/nrg2779.
- Crotti L, Johnson CN, Graf E, De Ferrari GM, Cuneo BF, Ovadia M, Papagiannis J, Feldkamp MD, Rathi SG, Kunic JD, Pedrazzini M, Wieland T, Lichtner P, Beckmann BM, Clark T, Shaffer C, Benson DW, Kääb S, Meitinger T, Strom TM, Chazin WJ, Schwartz PJ, George AL. Calmodulin mutations associated with recurrent cardiac arrest in infants. Circulation. 2013;127:1009–1017. doi: 10.1161/CIRCULATIONAHA.112.001216.
- Darpo B, Karnad DR, Badilini F, Florian J, Garnett CE, Kothari S, Panicker GK, Sarapa N. Are women more susceptible than men to drug-induced QT prolongation? Concentration-QTc modelling in a phase 1 study with oral rac-sotalol. British Journal of Clinical Pharmacology. 2014;77:522–531. doi: 10.1111/bcp.12201.
- Fermini B, Fossa AA. The impact of drug-induced QT interval prolongation on drug discovery and development. Nature Reviews Drug Discovery. 2003;2:439–447. doi: 10.1038/nrd1108.
- Funck-Brentano C, Jaillon P. Rate-corrected QT interval: techniques and limitations. The American Journal of Cardiology. 1993;72:17B–22. doi: 10.1016/0002-9149(93)90035-B.
- Gintant G. An evaluation of hERG current assay performance: Translating preclinical safety studies to clinical QT prolongation. Pharmacology & Therapeutics. 2011;129:109–119. doi: 10.1016/j.pharmthera.2010.08.008.
- Giorgi MA, Bolaños R, Gonzalez CD, Di Girolamo G. QT interval prolongation: preclinical and clinical testing arrhythmogenesis in drugs and regulatory implications. Current Drug Safety. 2010;5:54–57. doi: 10.2174/157488610789869148.
- Hulot JS, Démolis JL, Rivière R, Strabach S, Christin-Maitre S, Funck-Brentano C. Influence of endogenous oestrogens on QT interval duration. European Heart Journal. 2003;24:1663–1667. doi: 10.1016/S0195-668X(03)00436-6.
- Inoue H, Yamanaka S. The use of induced pluripotent stem cells in drug development. Clinical Pharmacology & Therapeutics. 2011;89:655–661. doi: 10.1038/clpt.2011.38.
- Itzhaki I, Maizels L, Huber I, Zwi-Dantsis L, Caspi O, Winterstern A, Feldman O, Gepstein A, Arbel G, Hammerman H, Boulos M, Gepstein L. Modelling the long QT syndrome with induced pluripotent stem cells. Nature. 2011;471:225–229. doi: 10.1038/nature09747.
- Kannankeril P, Roden DM, Darbar D. Drug-induced long QT syndrome. Pharmacological Reviews. 2010;62:760–781. doi: 10.1124/pr.110.003723.
- Kannankeril PJ, Roden DM, Norris KJ, Whalen SP, George AL, Murray KT. Genetic susceptibility to acquired long QT syndrome: pharmacologic challenge in first-degree relatives. Heart Rhythm. 2005;2:134–140. doi: 10.1016/j.hrthm.2004.10.039.
- Karakikes I, Senyei GD, Hansen J, Kong CW, Azeloglu EU, Stillitano F, Lieu DK, Wang J, Ren L, Hulot JS, Iyengar R, Li RA, Hajjar RJ. Small molecule-mediated directed differentiation of human embryonic stem cells toward ventricular cardiomyocytes. Stem Cells Translational Medicine. 2014;3:18–31. doi: 10.5966/sctm.2013-0110.
- Keller GA, Di Girolamo G. Prokinetic agents and QT prolongation: a familiar scene with new actors. Current Drug Safety. 2010;5:73–78. doi: 10.2174/157488610789869166.
- Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nature Methods. 2012;9:357–359. doi: 10.1038/nmeth.1923.
- Leonoudakis D, Conti LR, Anderson S, Radeke CM, McGuire LM, Adams ME, Froehner SC, Yates JR, Vandenberg CA. Protein trafficking and anchoring complexes revealed by proteomic analysis of inward rectifier potassium channel (Kir2.x)-associated proteins. Journal of Biological Chemistry. 2004;279:22331–22346. doi: 10.1074/jbc.M400285200.
- Levy DI, Cepaitis E, Wanderling S, Toth PT, Archer SL, Goldstein SA. The membrane protein MiRP3 regulates Kv4.2 channels in a KChIP-dependent manner. The Journal of Physiology. 2010;588:2657–2668. doi: 10.1113/jphysiol.2010.191395.
- Liang P, Lan F, Lee AS, Gong T, Sanchez-Freire V, Wang Y, Diecke S, Sallam K, Knowles JW, Wang PJ, Nguyen PK, Bers DM, Robbins RC, Wu JC. Drug screening using a library of human induced pluripotent stem cell-derived cardiomyocytes reveals disease-specific patterns of cardiotoxicity. Circulation. 2013;127:1677–1691. doi: 10.1161/CIRCULATIONAHA.113.001883.
- Liu Z, Liu X, Yu H, Pei J, Zhang Y, Gong J, Pu J. Common Variants in TRDN and CALM1 Are Associated with Risk of Sudden Cardiac Death in Chronic Heart Failure Patients in Chinese Han Population. PLOS One. 2015;10:e0132459. doi: 10.1371/journal.pone.0132459.
- Lundy SD, Zhu WZ, Regnier M, Laflamme MA. Structural and functional maturation of cardiomyocytes derived from human pluripotent stem cells. Stem Cells and Development. 2013;22:1991–2002. doi: 10.1089/scd.2012.0490.
- Maguy A, Hebert TE, Nattel S. Involvement of lipid rafts and caveolae in cardiac ion channel function. Cardiovascular Research. 2006;69:798–807. doi: 10.1016/j.cardiores.2005.11.013.
- Makkar RR. Female Gender as a Risk Factor for Torsades de Pointes Associated With Cardiovascular Drugs. JAMA. 1993;270:2590–2597. doi: 10.1001/jama.1993.03510210076031.
- Mann DA. Human induced pluripotent stem cell-derived hepatocytes for toxicology testing. Expert Opinion on Drug Metabolism & Toxicology. 2015;11:1–5. doi: 10.1517/17425255.2015.981523.
- McIntyre LM, Lopiano KK, Morse AM, Amin V, Oberg AL, Young LJ, Nuzhdin SV. RNA-seq: technical variability and sampling. BMC Genomics. 2011;12:293. doi: 10.1186/1471-2164-12-293.
- Moretti A, Bellin M, Welling A, Jung CB, Lam JT, Bott-Flügel L, Dorn T, Goedel A, Höhnke C, Hofmann F, Seyfarth M, Sinnecker D, Schömig A, Laugwitz KL. Patient-specific induced pluripotent stem-cell models for long-QT syndrome. New England Journal of Medicine. 2010;363:1397–1409. doi: 10.1056/NEJMoa0908679.
- Moss AJ, Robinson J. Clinical features of the idiopathic long QT syndrome. Circulation. 1992;85:I140–144.
- Nabi IR. Cavin fever: regulating caveolae. Nature Cell Biology. 2009;11:789–791. doi: 10.1038/ncb0709-789.
- Navarrete EG, Liang P, Lan F, Sanchez-Freire V, Simmons C, Gong T, Sharma A, Burridge PW, Patlolla B, Lee AS, Wu H, Beygui RE, Wu SM, Robbins RC, Bers DM, Wu JC. Screening drug-induced arrhythmia [corrected] using human induced pluripotent stem cell-derived cardiomyocytes and low-impedance microelectrode arrays. Circulation. 2013;128:S3–S13. doi: 10.1161/CIRCULATIONAHA.112.000570.
- Nozaki Y, Honda Y, Tsujimoto S, Watanabe H, Kunimatsu T, Funabashi H. Availability of human induced pluripotent stem cell-derived cardiomyocytes in assessment of drug potential for QT prolongation. Toxicology and Applied Pharmacology. 2014;278:72–77. doi: 10.1016/j.taap.2014.04.007.
- Odening KE, Koren G. How do sex hormones modify arrhythmogenesis in long QT syndrome? Sex hormone effects on arrhythmogenic substrate and triggered activity. Heart Rhythm. 2014;11:2107–2115. doi: 10.1016/j.hrthm.2014.06.023.
- Paulussen AD, Gilissen RA, Armstrong M, Doevendans PA, Verhasselt P, Smeets HJ, Schulze-Bahr E, Haverkamp W, Breithardt G, Cohen N, Aerssens J. Genetic variations of KCNQ1, KCNH2, SCN5A, KCNE1, and KCNE2 in drug-induced long QT syndrome patients. Journal of Molecular Medicine. 2004;82:182–188. doi: 10.1007/s00109-003-0522-z.
- Petropoulou E, Jamshidi Y, Behr ER. The genetics of pro-arrhythmic adverse drug reactions. British Journal of Clinical Pharmacology. 2014;77:618–625. doi: 10.1111/bcp.12208.
- Pfeufer A, Sanna S, Arking DE, Müller M, Gateva V, Fuchsberger C, Ehret GB, Orrú M, Pattaro C, Köttgen A, Perz S, Usala G, Barbalic M, Li M, Pütz B, Scuteri A, Prineas RJ, Sinner MF, Gieger C, Najjar SS, Kao WH, Mühleisen TW, Dei M, Happle C, Möhlenkamp S, Crisponi L, Erbel R, Jöckel KH, Naitza S, Steinbeck G, Marroni F, Hicks AA, Lakatta E, Müller-Myhsok B, Pramstaller PP, Wichmann HE, Schlessinger D, Boerwinkle E, Meitinger T, Uda M, Coresh J, Kääb S, Abecasis GR, Chakravarti A. Common variants at ten loci modulate the QT interval duration in the QTSCD Study. Nature Genetics. 2009;41:407–414. doi: 10.1038/ng.362.
- Rajab A, Straub V, McCann LJ, Seelow D, Varon R, Barresi R, Schulze A, Lucke B, Lützkendorf S, Karbasiyan M, Bachmann S, Spuler S, Schuelke M. Fatal cardiac arrhythmia and long-QT syndrome in a new form of congenital generalized lipodystrophy with muscle rippling (CGL4) due to PTRF-CAVIN mutations. PLoS Genetics. 2010;6:e1000874. doi: 10.1371/journal.pgen.1000874.
- Roden DM. Long QT syndrome: reduced repolarization reserve and the genetic link. Journal of Internal Medicine. 2006;259:59–69. doi: 10.1111/j.1365-2796.2005.01589.x.
- Roden DM. Cellular basis of drug-induced torsades de pointes. British Journal of Pharmacology. 2008a;154:1502–1507. doi: 10.1038/bjp.2008.238.
- Roden DM. Repolarization reserve: a moving target. Circulation. 2008b;118:981–982. doi: 10.1161/CIRCULATIONAHA.108.798918.
- Rolland T, Taşan M, Charloteaux B, Pevzner SJ, Zhong Q, Sahni N, Yi S, Lemmens I, Fontanillo C, Mosca R, Kamburov A, Ghiassian SD, Yang X, Ghamsari L, Balcha D, Begg BE, Braun P, Brehme M, Broly MP, Carvunis AR, Convery-Zupan D, Corominas R, Coulombe-Huntington J, Dann E, Dreze M, Dricot A, Fan C, Franzosa E, Gebreab F, Gutierrez BJ, Hardy MF, Jin M, Kang S, Kiros R, Lin GN, Luck K, MacWilliams A, Menche J, Murray RR, Palagi A, Poulin MM, Rambout X, Rasla J, Reichert P, Romero V, Ruyssinck E, Sahalie JM, Scholz A, Shah AA, Sharma A, Shen Y, Spirohn K, Tam S, Tejeda AO, Trigg SA, Twizere JC, Vega K, Walsh J, Cusick ME, Xia Y, Barabási AL, Iakoucheva LM, Aloy P, De Las Rivas J, Tavernier J, Calderwood MA, Hill DE, Hao T, Roth FP, Vidal M. A proteome-scale map of the human interactome network. Cell. 2014;159:1212–1226. doi: 10.1016/j.cell.2014.10.050.
- Sager PT, Gintant G, Turner JR, Pettit S, Stockbridge N. Rechanneling the cardiac proarrhythmia safety paradigm: a meeting report from the Cardiac Safety Research Consortium. American Heart Journal. 2014;167:292–300. doi: 10.1016/j.ahj.2013.11.004.
- Sarganas G, Garbe E, Klimpel A, Hering RC, Bronder E, Haverkamp W. Epidemiology of symptomatic drug-induced long QT syndrome and Torsade de Pointes in Germany. Europace. 2014;16:101–108. doi: 10.1093/europace/eut214.
- Schaaf S, Shibamiya A, Mewe M, Eder A, Stöhr A, Hirt MN, Rau T, Zimmermann WH, Conradi L, Eschenhagen T, Hansen A. Human engineered heart tissue as a versatile tool in basic research and preclinical toxicology. PLoS One. 2011;6:e26397. doi: 10.1371/journal.pone.0026397.
- Sinnecker D, Goedel A, Dorn T, Dirschinger RJ, Moretti A, Laugwitz KL. Modeling long-QT syndromes with iPS cells. Journal of Cardiovascular Translational Research. 2013;6:31–36. doi: 10.1007/s12265-012-9416-1.
- Sinnecker D, Laugwitz KL, Moretti A. Induced pluripotent stem cell-derived cardiomyocytes for drug development and toxicity testing. Pharmacology & Therapeutics. 2014;143:246–252. doi: 10.1016/j.pharmthera.2014.03.004.
- Soyka LF, Wirtz C, Spangenberg RB. Clinical safety profile of sotalol in patients with arrhythmias. The American Journal of Cardiology. 1990;65:74A–81. doi: 10.1016/0002-9149(90)90207-H.
- Terrenoire C, Wang K, Tung KW, Chung WK, Pass RH, Lu JT, Jean JC, Omari A, Sampson KJ, Kotton DN, Keller G, Kass RS. Induced pluripotent stem cells used to reveal drug actions in a long QT syndrome family with complex genetics. The Journal of General Physiology. 2013;141:61–72. doi: 10.1085/jgp.201210899.
- Trapnell C, Pachter L, Salzberg SL. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics. 2009;25:1105–1111. doi: 10.1093/bioinformatics/btp120.
- Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nature Biotechnology. 2010;28:511–515. doi: 10.1038/nbt.1621.
- Turnbull IC, Karakikes I, Serrao GW, Backeris P, Lee JJ, Xie C, Senyei G, Gordon RE, Li RA, Akar FG, Hajjar RJ, Hulot JS, Costa KD. Advancing functional engineered cardiac tissues toward a preclinical model of human myocardium. The FASEB Journal. 2014;28:644–654. doi: 10.1096/fj.13-228007.
- Wang Y, Liang P, Lan F, Wu H, Lisowski L, Gu M, Hu S, Kay MA, Urnov FD, Shinnawi R, Gold JD, Gepstein L, Wu JC. Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing. Journal of the American College of Cardiology. 2014;64:451–459. doi: 10.1016/j.jacc.2014.04.057.
- Yang P, Kanki H, Drolet B, Yang T, Wei J, Viswanathan PC, Hohnloser SH, Shimizu W, Schwartz PJ, Stanton M, Murray KT, Norris K, George AL, Roden DM. Allelic variants in long-QT disease genes in patients with drug-associated torsades de pointes. Circulation. 2002;105:1943–1948. doi: 10.1161/01.CIR.0000014448.19052.4C.
- Zanetti LA. Sotalol: a new class III antiarrhythmic agent. Clinical Pharmacy. 1993;12:883–891.
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