title: MicroRNAs as early toxicity signatures of doxorubicin in human-induced pluripotent stem cell-derived cardiomyocytes.
abstract: An in depth investigation at the genomic level is needed to identify early human-relevant cardiotoxicity biomarkers that are induced by drugs and environmental toxicants. The main objective of this study was to investigate the role of microRNAs (miRNAs) as cardiotoxicity biomarkers using human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) that were exposed to doxorubicin (DOX) as a "gold standard" cardiotoxicant. hiPSC-CMs were exposed to 156 nM DOX for 2 days or for 6 days of repeated exposure, followed by drug washout and incubation in drug-free culture medium up to day 14 after the onset of exposure. The induced miRNAs were profiled using miRNA microarrays, and the analysis of the data was performed using the miRWalk 2.0 and DAVID bioinformatics tools. DOX induced early deregulation of 14 miRNAs (10 up-regulated and 4 down-regulated) and persistent up-regulation of 5 miRNAs during drug washout. Computational miRNA gene target predictions suggested that several DOX-responsive miRNAs might regulate the mRNA expression of genes involved in cardiac contractile function. The hiPSC-CMs exposed to DOX in a range from 39 to 156 nM did not show a significant release of the cytotoxicity marker lactate dehydrogenase (LDH) compared to controls. Quantitative real-time PCR analyses confirmed the early deregulation of miR-187-3p, miR-182-5p, miR-486-3p, miR-486-5p, miR-34a-3p, miR-4423-3p, miR-34c-3p, miR-34c-5p and miR-1303, and also the prolonged up-regulation of miR-182-5p, miR-4423-3p and miR-34c-5p. Thus, we identified and validated miRNAs showing differential DOX-responsive expression before the occurrence of cytotoxicity markers such as LDH, and these miRNAs also demonstrated the significant involvement in heart failure in patients and animal models. These results suggest that the DOX-induced deregulated miRNAs in human CMs may be used as early sensitive cardiotoxicity biomarkers for screening potential drugs and environmental cardiotoxicants with a similar mechanism of action.
authors: Institute of Neurophysiology, Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany.
authors: Umesh
authors: U
authors: Chaudhari
authors: Institute of Neurophysiology, Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany.
authors: Harshal
authors: H
authors: Nemade
authors: Institute of Neurophysiology, Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany.
authors: John Antonydas
authors: JA
authors: Gaspar
authors: Institute of Neurophysiology, Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany.
authors: Jürgen
authors: J
authors: Hescheler
authors: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany.
authors: Jan G
authors: JG
authors: Hengstler
authors: Institute of Neurophysiology, Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany. a.sachinidis@uni-koeln.de.
authors: Agapios
authors: A
authors: Sachinidis
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D054316:Biomarkers, Pharmacological / Q000378:metabolism; D054715:Cardiotoxins / Q000633:toxicity*; D002454:Cell Differentiation; D002478:Cells, Cultured; D019295:Computational Biology; D004317:Doxorubicin / Q000633:toxicity*; D004353:Drug Evaluation, Preclinical; D015507:Drugs, Investigational / Q000009:adverse effects; D004784:Environmental Monitoring; D004785:Environmental Pollutants / Q000633:toxicity; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D007700:Kinetics; D035683:MicroRNAs / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008956:Models, Chemical; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; D054316:Biomarkers, Pharmacological; D054715:Cardiotoxins; D015507:Drugs, Investigational; D004785:Environmental Pollutants; D035683:MicroRNAs; D004317:Doxorubicin
keywords: Anthracyclines; Cardiotoxicity; Genomic biomarkers; miRNAs