title: High-throughput screening of tyrosine kinase inhibitor cardiotoxicity with human induced pluripotent stem cells.
abstract: Tyrosine kinase inhibitors (TKIs), despite their efficacy as anticancer therapeutics, are associated with cardiovascular side effects ranging from induced arrhythmias to heart failure. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), generated from 11 healthy individuals and 2 patients receiving cancer treatment, to screen U.S. Food and Drug Administration-approved TKIs for cardiotoxicities by measuring alterations in cardiomyocyte viability, contractility, electrophysiology, calcium handling, and signaling. With these data, we generated a "cardiac safety index" to reflect the cardiotoxicities of existing TKIs. TKIs with low cardiac safety indices exhibit cardiotoxicity in patients. We also derived endothelial cells (hiPSC-ECs) and cardiac fibroblasts (hiPSC-CFs) to examine cell type-specific cardiotoxicities. Using high-throughput screening, we determined that vascular endothelial growth factor receptor 2 (VEGFR2)/platelet-derived growth factor receptor (PDGFR)-inhibiting TKIs caused cardiotoxicity in hiPSC-CMs, hiPSC-ECs, and hiPSC-CFs. With phosphoprotein analysis, we determined that VEGFR2/PDGFR-inhibiting TKIs led to a compensatory increase in cardioprotective insulin and insulin-like growth factor (IGF) signaling in hiPSC-CMs. Up-regulating cardioprotective signaling with exogenous insulin or IGF1 improved hiPSC-CM viability during cotreatment with cardiotoxic VEGFR2/PDGFR-inhibiting TKIs. Thus, hiPSC-CMs can be used to screen for cardiovascular toxicities associated with anticancer TKIs, and the results correlate with clinical phenotypes. This approach provides unexpected insights, as illustrated by our finding that toxicity can be alleviated via cardioprotective insulin/IGF signaling.
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Arun
authors.identifier: 0000-0002-0607-5455
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Department of Pharmacology and Center for Pharmacogenomics, Northwestern University School of Medicine, Chicago, IL 60611, USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Burridge
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.|Graduate School of Biomedical Sciences, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
authors.forename: Wesley L
authors.initials: WL
authors.lastname: McKeithan
authors.affiliation: Department of Mechanical and Aerospace Engineering, University of California, San Diego, La Jolla, CA 92092, USA.
authors.forename: Ricardo
authors.identifier: 0000-0002-4206-3638
authors.initials: R
authors.lastname: Serrano
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Praveen
authors.initials: P
authors.lastname: Shukla
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Nazish
authors.initials: N
authors.lastname: Sayed
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Churko
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Tomoya
authors.identifier: 0000-0002-2696-046X
authors.initials: T
authors.lastname: Kitani
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Haodi
authors.initials: H
authors.lastname: Wu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Holmström
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Matsa
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Anusha
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Division of Oncology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Alice C
authors.initials: AC
authors.lastname: Fan
authors.affiliation: Department of Mechanical and Aerospace Engineering, University of California, San Diego, La Jolla, CA 92092, USA.
authors.forename: Juan C
authors.initials: JC
authors.lastname: Del Álamo
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Sean M
authors.initials: SM
authors.lastname: Wu
authors.affiliation: Division of Cardiovascular Medicine, Cardio-Oncology Program, Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN 37212, USA.
authors.forename: Javid J
authors.initials: JJ
authors.lastname: Moslehi
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
authors.forename: Mark
authors.identifier: 0000-0002-1430-2013
authors.initials: M
authors.lastname: Mercola
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA. joewu@stanford.edu.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.|Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Joseph C
authors.identifier: 0000-0002-6068-8041
authors.initials: JC
authors.lastname: Wu
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D066126:Cardiotoxicity / Q000473:pathology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology / Q000633:toxicity*; D012518:Sarcomeres / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007328:Insulin; D047428:Protein Kinase Inhibitors; D007334:Insulin-Like Growth Factor I; D040301:Vascular Endothelial Growth Factor Receptor-2