Validation of visualized transgenic zebrafish as a high throughput model to assay bradycardia related cardio toxicity risk candidates

Abstract

Drug-induced QT prolongation usually leads to torsade de pointes (TdP), thus for drugs in the early phase of development this risk should be evaluated. In the present study, we demonstrated a visualized transgenic zebrafish as an in vivo high-throughput model to assay the risk of drug-induced QT prolongation. Zebrafish larvae 48 h post-fertilization expressing green fluorescent protein in myocardium were incubated with compounds reported to induce QT prolongation or block the human ether-a-go-go-related gene (hERG) K⁺ current. The compounds sotalol, indapaminde, erythromycin, ofoxacin, levofloxacin, sparfloxacin and roxithromycin were additionally administrated by microinjection into the larvae yolk sac. The ventricle heart rate was recorded using the automatic monitoring system after incubation or microinjection. As a result, 14 out of 16 compounds inducing dog QT prolongation caused bradycardia in zebrafish. A similar result was observed with 21 out of 26 compounds which block hERG current. Among the 30 compounds which induced human QT prolongation, 25 caused bradycardia in this model. Thus, the risk of compounds causing bradycardia in this transgenic zebrafish correlated with that causing QT prolongation and hERG K⁺ current blockage in established models. The tendency that high logP values lead to high risk of QT prolongation in this model was indicated, and non-sensitivity of this model to antibacterial agents was revealed. These data suggest application of this transgenic zebrafish as a high-throughput model to screen QT prolongation-related cardio toxicity of the drug candidates.

ZeGlobalTox: An Innovative Approach to Address Organ Drug Toxicity Using Zebrafish

“…Interestingly, bradycardia was detected in 4 out of 6 drugs producing QTc prolongation in humans. This is consistent with results presented by Wen et al [57], which showed a correlation between drugs producing QTc prolongation (in dogs) and bradycardia in zebrafish. On the other hand, tachycardia was observed in d -(+)-glucose treated zebrafish larvae.…”

“…This seems certain for ethanol and paracetamol, since their human cardiotoxic impact is observed as a late effect after drug poisoning. In fact, paracetamol has been reported to have no impact on the heart rate in zebrafish larvae [50,57], to the point that Wen et al [54] included this drug as a negative cardiotoxic drug [57]. In summary, we support zebrafish as a powerful tool for predicting drug-induced cardiotoxic liabilities in humans, including typical repolarization and depolarization end-phenotypes such as QTc or EJC.…”

Pred‐hERG: A Novel web‐Accessible Computational Tool for Predicting Cardiac Toxicity

“…Testing a new bioactive molecule for hERG safety is mandatory and required by the US FDA. Experimental evaluation of binding affinity to hERG K + channel such as the “gold standard” patch‐clamp electrophysiology,6,7 the in vivo test on zebrafish,8 etc., are laborious, expensive, and time consuming 9. Therefore, there is a need in developing computational tools to reliably identify and filter out potential hERG blockers.…”

Tuning hERG Out: Antitarget QSAR Models for Drug Development

“…To identify the potential of a substance to delay ventricular repolarization and to estimate the risk for QT interval prolongation in humans, several experiments are conducted. Although new techniques have appeared in the last few years [11–13], conventional patch-clamp electrophysiology remains the “gold-standard.” In this test, the electric current passing through hERG channels expressed in cells, is measured [14]. Often, the cell lines utilized as an expression system for the hERG in studies include human embryonic kidney (HEK) 293 cells, Chinese hamster ovary (CHO) cells, or Xenopus laevis oocytes (XO) cells.…”