Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices
Abstract Acute block of hERG channels is the most common mechanism underlying drug-induced QTC prolongation and potentially fatal Torsade de Pointes arrhythmia. Updates to ICH E14 Q&As now allow for using negative nonclinical data, including hERG, to support QTC risk assessment in late-stage cli...
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Nature Portfolio
2025-08-01
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| Online Access: | https://doi.org/10.1038/s41598-025-15761-8 |
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| author | Claudia Alvarez Baron Jun Zhao Huimei Yu Ming Ren Nicolas Thiebaud Donglin Guo Giri Vegesna Cheng-Hui Hsiao Ryan DePalma Sabyasachy Mistry Isra Tariq Md Shadiqur Rashid Roni Omnia A. Ismaiel Murali K. Matta Vikram Patel Manni Mashaee Jose Vicente Lars Johannesen Jiansong Sheng Simon Hebeisen James Kramer Andrew Bruening-Wright Koji Nakano Hiroshi Matsukawa Jennifer Beck Pierson Wendy W. Wu |
| author_facet | Claudia Alvarez Baron Jun Zhao Huimei Yu Ming Ren Nicolas Thiebaud Donglin Guo Giri Vegesna Cheng-Hui Hsiao Ryan DePalma Sabyasachy Mistry Isra Tariq Md Shadiqur Rashid Roni Omnia A. Ismaiel Murali K. Matta Vikram Patel Manni Mashaee Jose Vicente Lars Johannesen Jiansong Sheng Simon Hebeisen James Kramer Andrew Bruening-Wright Koji Nakano Hiroshi Matsukawa Jennifer Beck Pierson Wendy W. Wu |
| author_sort | Claudia Alvarez Baron |
| collection | DOAJ |
| description | Abstract Acute block of hERG channels is the most common mechanism underlying drug-induced QTC prolongation and potentially fatal Torsade de Pointes arrhythmia. Updates to ICH E14 Q&As now allow for using negative nonclinical data, including hERG, to support QTC risk assessment in late-stage clinical development. To interpret the hERG results, understanding hERG assay reproducibility or hERG data variability is pivotal. Protocol and best practice recommendations have been provided with the goal of minimizing lab-to-lab data differences, but the impact remains unclear. To fill this knowledge gap, hERG data from a HESI-coordinated multi-laboratory study were leveraged. Using standardized protocol and following best practices for patch clamp studies, five laboratories tested 28 drugs using the manual patch clamp technique. Systematic differences in block potencies were observed for data generated by one laboratory for the first 21 drugs, and these differences disappeared for the last seven drugs. Exposure, pharmacological sensitivity of the cell lines, and cell/data qualities were ruled out as the factors underlying systematic differences. All laboratories retested two drugs and obtained results within 1.6X of the initial testings, except for another laboratory that obtained data for one drug that differed from its initial testing by 7.6X. Descriptive statistics and meta-analysis were applied to the dataset to estimate what the distribution in hERG block potencies would be if a laboratory were to test the same drug repeatedly. This measure, or hERG data variability, was ~ 5X. Based on these results, hERG block potency values within 5X of each other should not be considered different, since these values are within the natural data distribution of the hERG assay; laboratory-specific safety margin threshold may be required to account for systematic data differences. |
| format | Article |
| id | doaj-art-101fb0bef1454abdbb818bbca248877a |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-101fb0bef1454abdbb818bbca248877a2025-08-20T03:47:07ZengNature PortfolioScientific Reports2045-23222025-08-0115112010.1038/s41598-025-15761-8Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practicesClaudia Alvarez Baron0Jun Zhao1Huimei Yu2Ming Ren3Nicolas Thiebaud4Donglin Guo5Giri Vegesna6Cheng-Hui Hsiao7Ryan DePalma8Sabyasachy Mistry9Isra Tariq10Md Shadiqur Rashid Roni11Omnia A. Ismaiel12Murali K. Matta13Vikram Patel14Manni Mashaee15Jose Vicente16Lars Johannesen17Jiansong Sheng18Simon Hebeisen19James Kramer20Andrew Bruening-Wright21Koji Nakano22Hiroshi Matsukawa23Jennifer Beck Pierson24Wendy W. Wu25Division of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Cardiology and Nephrology, Office of Cardiology, Hematology, Endocrinology and Nephrology, Office of New Drugs, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Cardiology and Nephrology, Office of Cardiology, Hematology, Endocrinology and Nephrology, Office of New Drugs, Center for Drug Evaluation and Research, US Food and Drug AdministrationDivision of Cardiology and Nephrology, Office of Cardiology, Hematology, Endocrinology and Nephrology, Office of New Drugs, Center for Drug Evaluation and Research, US Food and Drug AdministrationCiPA LabB’SYS GmbHCharles River LaboratoriesCharles River LaboratoriesDrug Safety Testing Center Co., Ltd., Higashimatsuyama LaboratoriesDrug Safety Testing Center Co., Ltd., Higashimatsuyama LaboratoriesHealth and Environmental Science InstituteDivision of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug AdministrationAbstract Acute block of hERG channels is the most common mechanism underlying drug-induced QTC prolongation and potentially fatal Torsade de Pointes arrhythmia. Updates to ICH E14 Q&As now allow for using negative nonclinical data, including hERG, to support QTC risk assessment in late-stage clinical development. To interpret the hERG results, understanding hERG assay reproducibility or hERG data variability is pivotal. Protocol and best practice recommendations have been provided with the goal of minimizing lab-to-lab data differences, but the impact remains unclear. To fill this knowledge gap, hERG data from a HESI-coordinated multi-laboratory study were leveraged. Using standardized protocol and following best practices for patch clamp studies, five laboratories tested 28 drugs using the manual patch clamp technique. Systematic differences in block potencies were observed for data generated by one laboratory for the first 21 drugs, and these differences disappeared for the last seven drugs. Exposure, pharmacological sensitivity of the cell lines, and cell/data qualities were ruled out as the factors underlying systematic differences. All laboratories retested two drugs and obtained results within 1.6X of the initial testings, except for another laboratory that obtained data for one drug that differed from its initial testing by 7.6X. Descriptive statistics and meta-analysis were applied to the dataset to estimate what the distribution in hERG block potencies would be if a laboratory were to test the same drug repeatedly. This measure, or hERG data variability, was ~ 5X. Based on these results, hERG block potency values within 5X of each other should not be considered different, since these values are within the natural data distribution of the hERG assay; laboratory-specific safety margin threshold may be required to account for systematic data differences.https://doi.org/10.1038/s41598-025-15761-8Cardiac safetyDelayed ventricular repolarizationExperimental uncertaintyConcentration-inhibitionAssay reproducibilitySafety margin |
| spellingShingle | Claudia Alvarez Baron Jun Zhao Huimei Yu Ming Ren Nicolas Thiebaud Donglin Guo Giri Vegesna Cheng-Hui Hsiao Ryan DePalma Sabyasachy Mistry Isra Tariq Md Shadiqur Rashid Roni Omnia A. Ismaiel Murali K. Matta Vikram Patel Manni Mashaee Jose Vicente Lars Johannesen Jiansong Sheng Simon Hebeisen James Kramer Andrew Bruening-Wright Koji Nakano Hiroshi Matsukawa Jennifer Beck Pierson Wendy W. Wu Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices Scientific Reports Cardiac safety Delayed ventricular repolarization Experimental uncertainty Concentration-inhibition Assay reproducibility Safety margin |
| title | Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices |
| title_full | Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices |
| title_fullStr | Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices |
| title_full_unstemmed | Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices |
| title_short | Multi-laboratory comparisons of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices |
| title_sort | multi laboratory comparisons of manual patch clamp herg data generated using standardized protocols and following ich s7b q a 2 1 best practices |
| topic | Cardiac safety Delayed ventricular repolarization Experimental uncertainty Concentration-inhibition Assay reproducibility Safety margin |
| url | https://doi.org/10.1038/s41598-025-15761-8 |
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