Multi-parameter in vitro toxicity testing of crizotinib, sunitinib, erlotinib, and nilotinib in human cardiomyocytes
Kimberly R Doherty1, Robert L Wappel, Dominique R Talbert
1Quintiles, 777 Oakmont Lane Suite 100, Westmont, IL 60559, USA.
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.
Insights
A multi-parameter test panel accurately predicts tyrosine kinase inhibitor (TKi) cardiotoxicity. This approach identifies drug-induced cardiomyocyte damage, improving safety during early drug development.
Area of Science:
- Cardiovascular Pharmacology
- Oncology Drug Development
- Toxicology
Background:
- Tyrosine kinase inhibitors (TKIs) are vital cancer therapeutics, but can cause cardiotoxicity not fully predicted by current preclinical models.
- Existing preclinical assessments, including animal studies and hERG channel inhibition assays, have limitations in predicting clinical cardiotoxicity.
- There is a critical need for improved methods to assess TKI-induced cardiac adverse events early in drug development.
Purpose of the Study:
- To evaluate a comprehensive in vitro test panel for predicting TKI cardiotoxicity.
- To assess the impact of FDA-approved TKIs on cardiomyocyte cellular, molecular, and electrophysiological functions.
- To determine if a multi-parameter approach can accurately characterize drug-induced cardiac damage.
Main Methods:
- Human cardiomyocytes were treated with four FDA-approved TKIs (crizotinib, sunitinib, nilotinib, erlotinib).
- Assessed endpoints included cell viability, apoptosis, reactive oxygen species (ROS) generation, metabolic status, impedance, and ion channel function.
- Utilized a multi-parameter test panel integrating cellular, molecular, and electrophysiological measurements.
Main Results:
- Three cardiotoxic TKIs (crizotinib, sunitinib, nilotinib) induced significant cellular and molecular damage in vitro.
- Crizotinib, sunitinib, and nilotinib demonstrated unique toxicity profiles, including increased ROS, apoptosis, metabolic disruption, and ion channel blockade (including hERG).
- The relatively cardiac-safe TKI, erlotinib, showed minimal adverse effects on cardiomyocyte health.
Conclusions:
- A multi-parameter in vitro testing strategy robustly characterizes TKI-induced cardiotoxicity.
- This comprehensive approach offers a more accurate prediction of cardiac adverse events compared to traditional methods.
- The findings support the integration of this panel into early drug development to enhance cardiovascular safety assessments for TKIs.
