Electrophysiological analysis of mammalian cells expressing hERG using automated 384-well-patch-clamp

BackgroundAn in vitro electrophysiological assay system, which can assess compound effects and thus show cardiotoxicity including arrhythmia risks of test drugs, is an essential method in the field of drug development and toxicology.MethodsIn this study, high-throughput electrophysiological recordings of human embryonic kidney (HEK 293) cells and Chinese hamster ovary (CHO) cells stably expressing human ether-a-go-go related gene (hERG) were performed utilizing an automated 384-well-patch-clamp system, which records up to 384 cells simultaneously. hERG channel inhibition, which is closely related to a drug-induced QT prolongation and is increasing the risk of sudden cardiac death, was investigated in the high-throughput screening patch-clamp system.ResultsIn the automated patch-clamp measurements performed here, Kv currents were investigated with high efficiency. Various hERG channel blockers showed concentration-dependent inhibition, the 50 % inhibitory concentrations (IC50) of those blockers were in good agreement with previous reports.ConclusionsThe high-throughput patch-clamp system has a high potential in the field of pharmacology, toxicology, and cardiac physiology, and will contribute to the acceleration of pharmaceutical drug development and drug safety testing.

[1]  Masayuki Yamato,et al.  Concise Review: Cell Therapy and Tissue Engineering for Cardiovascular Disease , 2012, Stem cells translational medicine.

[2]  Raymond L Woosley,et al.  Comparative Evaluation of HERG Currents and QT Intervals following Challenge with Suspected Torsadogenic and Nontorsadogenic Drugs , 2006, Journal of Pharmacology and Experimental Therapeutics.

[3]  J. Weirich,et al.  Rate-dependence of antiarrhythmic and proarrhythmic properties of class I and class III antiarrhythmic drugs , 1998, Basic Research in Cardiology.

[4]  Marc Peschanski,et al.  Human pluripotent stem cells in drug discovery and predictive toxicology. , 2010, Biochemical Society transactions.

[5]  Masayuki Yamato,et al.  Creation of human cardiac cell sheets using pluripotent stem cells. , 2012, Biochemical and biophysical research communications.

[6]  Gea-Ny Tseng,et al.  IKr: The hERG Channel , 2001 .

[7]  Akinori Noma,et al.  Molecular Characterization of the Hyperpolarization-activated Cation Channel in Rabbit Heart Sinoatrial Node* , 1999, The Journal of Biological Chemistry.

[8]  R. Schwinger,et al.  Effect of inotropic interventions on the force-frequency relation in the human heart , 1998, Basic Research in Cardiology.

[9]  T. Okano,et al.  Simple suspension culture system of human iPS cells maintaining their pluripotency for cardiac cell sheet engineering , 2015, Journal of tissue engineering and regenerative medicine.

[10]  J. Warmke,et al.  A family of potassium channel genes related to eag in Drosophila and mammals. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[11]  James Kramer,et al.  Comparison of block among cloned cardiac potassium channels by non-antiarrhythmic drugs , 2001 .

[12]  P. Emery,et al.  TRPC channel activation by extracellular thioredoxin , 2008, Nature.

[13]  L. Possani,et al.  Toxin modulators and blockers of hERG K(+) channels. , 2012, Toxicon : official journal of the International Society on Toxinology.

[14]  Mark L Dallas,et al.  Robotic multiwell planar patch-clamp for native and primary mammalian cells , 2009, Nature Protocols.

[15]  G. Robertson,et al.  HERG, a human inward rectifier in the voltage-gated potassium channel family. , 1995, Science.

[16]  M. Tada,et al.  Progressive maturation in contracting cardiomyocytes derived from human embryonic stem cells: Qualitative effects on electrophysiological responses to drugs. , 2010, Stem cell research.

[17]  I. Kola,et al.  Can the pharmaceutical industry reduce attrition rates? , 2004, Nature Reviews Drug Discovery.

[18]  Carlo Napolitano,et al.  Induced pluripotent stem cell-derived cardiomyocytes in studies of inherited arrhythmias. , 2013, The Journal of clinical investigation.

[19]  A J Camm,et al.  Arrhythmogenic mechanisms of non‐sedating antihistamines , 1999, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[20]  B. Sakmann,et al.  Single-channel currents recorded from membrane of denervated frog muscle fibres , 1976, Nature.

[21]  G. Salama,et al.  High-purity enrichment of functional cardiovascular cells from human iPS cells. , 2012, Cardiovascular research.

[22]  A. Brown,et al.  Variability in the measurement of hERG potassium channel inhibition: effects of temperature and stimulus pattern. , 2004, Journal of pharmacological and toxicological methods.

[23]  G. Tseng.  I(Kr): the hERG channel. , 2001, Journal of molecular and cellular cardiology.

[24]  H. Katus,et al.  Anesthetic drug midazolam inhibits cardiac human ether-à-go-go-related gene channels: mode of action , 2015, Drug design, development and therapy.

[25]  A. Verkhratsky,et al.  History of electrophysiology and the patch clamp. , 2014, Methods in molecular biology.

[26]  Ralf Kettenhofen,et al.  State-of-the-Art Automated Patch Clamp Devices: Heat Activation, Action Potentials, and High Throughput in Ion Channel Screening , 2011, Front. Pharmacol..

[27]  Jeffrey J Clare,et al.  Targeting ion channels for drug discovery. , 2010, Discovery medicine.

[28]  Stefan A. Mann,et al.  hERG K(+) channels: structure, function, and clinical significance. , 2012, Physiological reviews.

[29]  H. Nakaya.  [Electropharmacological assessment of the risk of drug-induced long-QT syndrome using native cardiac cells and cultured cells expressing HERG channels]. , 2003, Nihon yakurigaku zasshi. Folia pharmacologica Japonica.