Multi-parameter in vitro toxicity testing of crizotinib, sunitinib, erlotinib, and nilotinib in human cardiomyocytes

Toxicol Appl Pharmacol. 2013 Oct 1;272(1):245-55. doi: 10.1016/j.taap.2013.04.027. Epub 2013 May 21.

Abstract

Tyrosine kinase inhibitors (TKi) have greatly improved the treatment and prognosis of multiple cancer types. However, unexpected cardiotoxicity has arisen in a subset of patients treated with these agents that was not wholly predicted by pre-clinical testing, which centers around animal toxicity studies and inhibition of the human Ether-à-go-go-Related Gene (hERG) channel. Therefore, we sought to determine whether a multi-parameter test panel assessing the effect of drug treatment on cellular, molecular, and electrophysiological endpoints could accurately predict cardiotoxicity. We examined how 4 FDA-approved TKi agents impacted cell viability, apoptosis, reactive oxygen species (ROS) generation, metabolic status, impedance, and ion channel function in human cardiomyocytes. The 3 drugs clinically associated with severe cardiac adverse events (crizotinib, sunitinib, nilotinib) all proved to be cardiotoxic in our in vitro tests while the relatively cardiac-safe drug erlotinib showed only minor changes in cardiac cell health. Crizotinib, an ALK/MET inhibitor, led to increased ROS production, caspase activation, cholesterol accumulation, disruption in cardiac cell beat rate, and blockage of ion channels. The multi-targeted TKi sunitinib showed decreased cardiomyocyte viability, AMPK inhibition, increased lipid accumulation, disrupted beat pattern, and hERG block. Nilotinib, a second generation Bcr-Abl inhibitor, led to increased ROS generation, caspase activation, hERG block, and an arrhythmic beat pattern. Thus, each drug showed a unique toxicity profile that may reflect the multiple mechanisms leading to cardiotoxicity. This study demonstrates that a multi-parameter approach can provide a robust characterization of drug-induced cardiomyocyte damage that can be leveraged to improve drug safety during early phase development.

Keywords: Cardiomyocytes; Cardiotoxicity; Drug evaluation; In vitro; Preclinical; Tyrosine kinase inhibitors.

MeSH terms

  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cholesterol / metabolism
  • Crizotinib
  • ERG1 Potassium Channel
  • Enzyme Activation / drug effects
  • Erlotinib Hydrochloride
  • Ether-A-Go-Go Potassium Channels / biosynthesis
  • Ether-A-Go-Go Potassium Channels / genetics
  • Humans
  • Indoles / toxicity
  • Ion Channels / drug effects
  • Lipids / biosynthesis
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / ultrastructure
  • Patch-Clamp Techniques
  • Pluripotent Stem Cells / drug effects
  • Protein Kinase Inhibitors / toxicity*
  • Protein-Tyrosine Kinases / antagonists & inhibitors*
  • Pyrazoles / toxicity
  • Pyridines / toxicity
  • Pyrimidines / toxicity
  • Pyrroles / toxicity
  • Quinazolines / toxicity
  • RNA / biosynthesis
  • RNA / isolation & purification
  • Reactive Oxygen Species / metabolism
  • Real-Time Polymerase Chain Reaction
  • Sunitinib

Substances

  • ERG1 Potassium Channel
  • Ether-A-Go-Go Potassium Channels
  • Indoles
  • Ion Channels
  • KCNH2 protein, human
  • Lipids
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyridines
  • Pyrimidines
  • Pyrroles
  • Quinazolines
  • Reactive Oxygen Species
  • Crizotinib
  • RNA
  • Cholesterol
  • Erlotinib Hydrochloride
  • Protein-Tyrosine Kinases
  • Caspase 3
  • Caspase 7
  • nilotinib
  • Sunitinib