title: Use of human stem cell derived cardiomyocytes to examine sunitinib mediated cardiotoxicity and electrophysiological alterations.
abstract: Sunitinib, an oral tyrosine kinase inhibitor approved to treat advanced renal cell carcinoma and gastrointestinal stroma tumor, is associated with clinical cardiac toxicity. Although the precise mechanism of sunitinib cardiotoxicity is not known, both the key metabolic energy regulator, AMP-activated protein kinase (AMPK), and ribosomal S 6 kinase (RSK) have been hypothesized as causative, albeit based on rodent models. To study the mechanism of sunitinib-mediated cardiotoxicity in a human model, induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) having electrophysiological and contractile properties of native cardiac tissue were investigated. Sunitinib was cardiotoxic in a dose-dependent manner with an IC₅₀ in the low micromolar range, observed by a loss of cellular ATP, an increase in oxidized glutathione, and induction of apoptosis in iPSC-CMs. Pretreatment of iPSC-CMs with AMPK activators AICAR or metformin, increased the phosphorylation of pAMPK-T172 and pACC-S79, but only marginally attenuated sunitinib mediated cell death. Furthermore, additional inhibitors of AMPK were not directly cytotoxic to iPSC-CMs up to 250 μM concentrations. Inhibition of RSK with a highly specific, irreversible, small molecule inhibitor (RSK-FMK-MEA) did not induce cytotoxicity in iPSC-CMs below 250 μM. Extensive electrophysiological analysis of sunitinib and RSK-FMK-MEA mediated conduction effects were performed. Taken together, these findings suggest that inhibition of AMPK and RSK are not a major component of sunitinib-induced cardiotoxicity. Although the exact mechanism of cardiotoxicity of sunitinib is not known, it is likely due to inhibition of multiple kinases simultaneously. These data highlight the utility of human iPSC-CMs in investigating the potential molecular mechanisms underlying drug-induced cardiotoxicity.
authors: Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110, USA.
authors: J D
authors: JD
authors: Cohen
authors: J E
authors: JE
authors: Babiarz
authors: R M
authors: RM
authors: Abrams
authors: L
authors: L
authors: Guo
authors: S
authors: S
authors: Kameoka
authors: E
authors: E
authors: Chiao
authors: J
authors: J
authors: Taunton
authors: K L
authors: KL
authors: Kolaja
mesh_terms: D000091162:AMP-Activated Protein Kinase Kinases; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D055724:Electrophysiological Phenomena / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000633:toxicity*; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000502:physiology; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000502:physiology; D011494:Protein Kinases / Q000378:metabolism; D011758:Pyrroles / Q000633:toxicity*; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000037:antagonists & inhibitors / Q000378:metabolism; D000077210:Sunitinib
chemical_list: D007211:Indoles; D011758:Pyrroles; D011494:Protein Kinases; D020794:Receptor Protein-Tyrosine Kinases; D038744:Ribosomal Protein S6 Kinases, 90-kDa; C069330:ribosomal protein S6 kinase, 90kDa, polypeptide 3; D000091162:AMP-Activated Protein Kinase Kinases; D000077210:Sunitinib