Phenotype-Based High-Throughput Classification of Long QT Syndrome Subtypes Using Human Induced Pluripotent Stem Cells

Summary For long QT syndrome (LQTS), recent progress in genome-sequencing technologies enabled the identification of rare genomic variants with diagnostic, prognostic, and therapeutic implications. However, pathogenic stratification of the identified variants remains challenging, especially in variants of uncertain significance. This study aimed to propose a phenotypic cell-based diagnostic assay for identifying LQTS to recognize pathogenic variants in a high-throughput manner suitable for screening. We investigated the response of LQT2-induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs) following IKr blockade using a multi-electrode array, finding that the response to IKr blockade was significantly smaller than in Control-iPSC-CMs. Furthermore, we found that LQT1-iPSC-CMs and LQT3-iPSC-CMs could be distinguished from Control-iPSC-CMs by IKs blockade and INa blockade, respectively. This strategy might be helpful in compensating for the shortcomings of genetic testing of LQTS patients.

[1]  James A Thomson,et al.  High purity human-induced pluripotent stem cell-derived cardiomyocytes: electrophysiological properties of action potentials and ionic currents. , 2011, American journal of physiology. Heart and circulatory physiology.

[2]  S. Yamanaka,et al.  The Use of Induced Pluripotent Stem Cells in Drug Development , 2011, Clinical pharmacology and therapeutics.

[3]  M. Suematsu,et al.  Distinct metabolic flow enables large-scale purification of mouse and human pluripotent stem cell-derived cardiomyocytes. , 2013, Cell stem cell.

[4]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells From Adult Human Fibroblasts by Defined Factors , 2008 .

[5]  Yasunari Kanda,et al.  Improvement of acquisition and analysis methods in multi-electrode array experiments with iPS cell-derived cardiomyocytes. , 2015, Journal of pharmacological and toxicological methods.

[6]  G. Ewald,et al.  Reduced response to IKr blockade and altered hERG1a/1b stoichiometry in human heart failure. , 2016, Journal of molecular and cellular cardiology.

[7]  C. Backer,et al.  Characterization of inwardly rectifying K+ channel in human cardiac myocytes. Alterations in channel behavior in myocytes isolated from patients with idiopathic dilated cardiomyopathy. , 1995, Circulation.

[8]  G. Breithardt,et al.  Life-threatening Arrhythmias Genotype-phenotype Correlation in the Long-qt Syndrome : Gene-specific Triggers for Genotype-phenotype Correlation in the Long-qt Syndrome Gene-specific Triggers for Life-threatening Arrhythmias , 2022 .

[9]  G. Bett,et al.  Ancillary subunits and stimulation frequency determine the potency of chromanol 293B block of the KCNQ1 potassium channel , 2006, The Journal of physiology.

[10]  M. Ackerman,et al.  Genotype- and phenotype-guided management of congenital long QT syndrome. , 2013, Current problems in cardiology.

[11]  C. Tracy,et al.  HRS/EHRA/APHRS expert consensus statement on the diagnosis and management of patients with inherited primary arrhythmia syndromes: document endorsed by HRS, EHRA, and APHRS in May 2013 and by ACCF, AHA, PACES, and AEPC in June 2013. , 2013, Heart rhythm.

[12]  Sean P. Palecek,et al.  Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions , 2012, Nature Protocols.

[13]  J. Stockman.  Genetic Testing in the Long QT Syndrome: Development and Validation of an Efficient Approach to Genotyping in Clinical Practice , 2007 .

[14]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[15]  Kevin E. Healy,et al.  Calcium Transients Closely Reflect Prolonged Action Potentials in iPSC Models of Inherited Cardiac Arrhythmia , 2014, Stem cell reports.

[16]  S. Yuasa,et al.  Induced Pluripotent Stem Cells in Cardiovascular Medicine , 2011, Stem cells international.

[17]  P. Gee,et al.  Efficient genomic correction methods in human iPS cells using CRISPR-Cas9 system. , 2016, Methods.

[18]  Jennifer L. Robinson,et al.  Long QT Syndrome. , 2003, JAMA.

[19]  M. Horie,et al.  Complex aberrant splicing in the induced pluripotent stem cell-derived cardiomyocytes from a patient with long QT syndrome carrying KCNQ1-A344Aspl mutation. , 2018, Heart rhythm.

[20]  S. Priori,et al.  Genetic testing in the long QT syndrome: development and validation of an efficient approach to genotyping in clinical practice. , 2005, JAMA.

[21]  S. Priori,et al.  Effectiveness and limitations of beta-blocker therapy in congenital long-QT syndrome. , 2000, Circulation.

[22]  Michael J Ackerman,et al.  Impact of genetics on the clinical management of channelopathies. , 2013, Journal of the American College of Cardiology.

[23]  H. Wellens,et al.  Novel Insights in the Congenital Long QT Syndrome , 2002, Annals of Internal Medicine.

[24]  M. Horie.  Molecular genetics have opened a new era for arrhythmia research, but also Pandora׳s box? , 2016, Journal of arrhythmia.

[25]  S. Priori,et al.  Association of Long QT Syndrome Loci and Cardiac Events Among Patients Treated With β-Blockers , 2004 .

[26]  A. V. van Ginneken,et al.  Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual IK1 , 2015, Front. Physiol..

[27]  N. Terada,et al.  Concise Review: Induced Pluripotent Stem Cell Research in the Era of Precision Medicine , 2017, Stem cells.

[28]  Michael J Ackerman,et al.  Genetic Testing for Long-QT Syndrome: Distinguishing Pathogenic Mutations From Benign Variants , 2009, Circulation.

[29]  Qinlian Zhou,et al.  Electronic "expression" of the inward rectifier in cardiocytes derived from human-induced pluripotent stem cells. , 2013, Heart rhythm.