title: Multi-parameter in vitro toxicity testing of crizotinib, sunitinib, erlotinib, and nilotinib in human cardiomyocytes.
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.
authors.affiliation: Quintiles, 777 Oakmont Lane Suite 100, Westmont, IL 60559, USA. Electronic address: kimberly.doherty@quintiles.com.
authors.forename: Kimberly R
authors.initials: KR
authors.lastname: Doherty
authors.forename: Robert L
authors.initials: RL
authors.lastname: Wappel
authors.forename: Dominique R
authors.initials: DR
authors.lastname: Talbert
authors.forename: Patricia B
authors.initials: PB
authors.lastname: Trusk
authors.forename: Diarmuid M
authors.initials: DM
authors.lastname: Moran
authors.forename: James W
authors.initials: JW
authors.lastname: Kramer
authors.forename: Arthur M
authors.initials: AM
authors.lastname: Brown
authors.forename: Scott A
authors.initials: SA
authors.lastname: Shell
authors.forename: Sarah
authors.initials: S
authors.lastname: Bacus
mesh_terms: D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D000077547:Crizotinib; D000072237:ERG1 Potassium Channel; D004789:Enzyme Activation / Q000187:drug effects; D000069347:Erlotinib Hydrochloride; D051638:Ether-A-Go-Go Potassium Channels / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007211:Indoles / Q000633:toxicity; D007473:Ion Channels / Q000187:drug effects; D008055:Lipids / Q000096:biosynthesis; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques; D039904:Pluripotent Stem Cells / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000633:toxicity*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011720:Pyrazoles / Q000633:toxicity; D011725:Pyridines / Q000633:toxicity; D011743:Pyrimidines / Q000633:toxicity; D011758:Pyrroles / Q000633:toxicity; D011799:Quinazolines / Q000633:toxicity; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000077210:Sunitinib
chemical_list: D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D007211:Indoles; D007473:Ion Channels; C000606913:KCNH2 protein, human; D008055:Lipids; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D011743:Pyrimidines; D011758:Pyrroles; D011799:Quinazolines; D017382:Reactive Oxygen Species; D000077547:Crizotinib; D012313:RNA; D002784:Cholesterol; D000069347:Erlotinib Hydrochloride; D011505:Protein-Tyrosine Kinases; D053148:Caspase 3; D053179:Caspase 7; C498826:nilotinib; D000077210:Sunitinib
keywords: Cardiomyocytes; Cardiotoxicity; Drug evaluation; In vitro; Preclinical; Tyrosine kinase inhibitors