Assessment of acute and chronic toxicity of doxorubicin in human induced pluripotent stem cell-derived cardiomyocytes

• Published in: Toxicology in vitro Vol. 42; pp. 182 - 190
• Main Authors: Louisse, Jochem, Wüst, Rob C.I., Pistollato, Francesca, Palosaari, Taina, Barilari, Manuela, Macko, Peter, Bremer, Susanne, Prieto, Pilar
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2017; Elsevier Science Ltd
• Subjects: Acute toxicity; Antibiotics, Antineoplastic - toxicity; Biocompatibility; Biomarkers; Calcium; Calcium (mitochondrial); Calcium - metabolism; Cardiomyocytes; Cardiotoxicity; Cell number; Cell survival; Cells, Cultured; Chemotherapy; Chronic effects; Chronic toxicity; Density; Doxorubicin; Doxorubicin - toxicity; High content imaging; Humans; Imaging techniques; In vitro; In vivo methods and tests; Induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Membrane potential; Membrane Potential, Mitochondrial - drug effects; Mitochondria; Mitochondria, Heart - drug effects; Mitochondria, Heart - physiology; Multi electrode array; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - physiology; Organic chemistry; Pluripotency; Stem cells; Superoxide; Superoxides - metabolism; Toxicity; MEA; CDI; Multi electrode array; DETECTIVE; FCCP; Cardiotoxicity; Doxorubicin; In vitro; Mitochondria; Induced pluripotent stem cells; ROS; hiPSC-CMs; SEURAT-1; High content imaging
• ISSN: 0887-2333; 1879-3177; 1879-3177
• DOI: 10.1016/j.tiv.2017.04.023

The present study assesses acute and chronic toxicity of doxorubicin in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), with the aim to obtain in vitro biomarkers that can be used as readouts to predict in vivo cardiotoxicity. Possible acute toxicity was investigated by assessing effects on the beating rate and the field potential duration (FPD) of doxorubicin-exposed cardiomyocytes by measuring electrical activity using multi-electrode array (MEA) analyses. No effects on the beating rate and FPD were found at concentrations up to 6μM, whereas at 12μM no electrical activity was recorded, indicating that the cardiomyocytes stopped beating. Acute and chronic effects of doxorubicin on mitochondria, which have been reported to be affected in doxorubicin-induced cardiotoxicity, were assessed using high content imaging techniques. To this end hiPSC-CMs were exposed to 150 or 300nM doxorubicin using both single dosing (3h and 2days) and repetitive dosing (3 times, of 2days each), including washout studies to assess delayed effects (assessment at day 14) and effects on cell number, mitochondrial density, mitochondrial membrane potential, mitochondrial superoxide levels and mitochondrial calcium levels were assessed. No effects of doxorubicin were found on mitochondrial density and mitochondrial superoxide levels, whereas doxorubicin reduced cell survival and slightly altered mitochondrial membrane potential and mitochondrial calcium levels, which was most profound in the washout studies. Altogether, the results of the present study show that concentrations of doxorubicin in the micromolar range were required to affect electrical activity of hiPSC-CMs, whereas nanomolar concentrations already affected cell viability and caused mitochondrial disturbances. Integration of these data with other in vitro data may enable the selection of a series of in vitro biomarkers that can be used as readouts to screen chemicals for possible cardiotoxicity. •The effects of doxorubicin on human stem cell-derived cardiomyocytes were studied.•Exposure consisted of single and repetitive dosing, including washout studies.•Doxorubicin mainly affected survival of cardiomyocytes.•Doxorubicin slightly affected mitochondrial membrane potential and calcium levels.