title: Cellular impedance assays for predictive preclinical drug screening of kinase inhibitor cardiovascular toxicity.
abstract: Cardiovascular (CV) toxicity is a leading contributor to drug attrition. Implementing earlier testing has successfully reduced human Ether-à-go-go-Related Gene-related arrhythmias. How- ever, analogous assays targeting functional CV effects remain elusive. Demand to address this gap is particularly acute for kinase inhibitors (KIs) that suffer frequent CV toxicity. The drug class also presents some particularly challenging requirements for assessing functional CV toxicity. Specifically, an assay must sense a downstream response that integrates diverse kinase signaling pathways. In addition, sufficient throughput is essential for handling inherent KI nonselectivity. A new opportunity has emerged with cellular impedance technology, which detects spontaneous beating cardiomyocytes. Impedance assays sense morphology changes downstream of cardiomyocyte contraction. To evaluate cardiomyocyte impedance assays for KI screening, we investigated two distinct KI classes where CV toxicity was discovered late and target risks remain unresolved. Microtubule-associated protein/microtubule affinity regulating kinase (MARK) inhibitors decrease blood pressure in dogs, whereas checkpoint kinase (Chk) inhibitors (AZD7762, SCH900776) exhibit dose-limiting CV toxicities in clinical trials. These in vivo effects manifested in vitro as cardiomyocyte beat cessation. MARK effects were deemed mechanism associated because beat inhibition potencies correlated with kinase inhibition, and gene knockdown and microtubule-targeting agents suppressed beating. MARK inhibitor impedance and kinase potencies aligned with rat blood pressure effects. Chk inhibitor effects were judged off-target because Chk and beat inhibition potencies did not correlate and knockdowns did not alter beating. Taken together, the data demonstrate that cardiomyocyte impedance assays can address three unmet needs-detecting KI functional cardiotoxicity in vitro, determining mechanism of action, and supporting safety structure-activity relationships.
authors.affiliation: Molecular Toxicology,Global Safety Assessment, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, USA.
authors.forename: Sarah D
authors.initials: SD
authors.lastname: Lamore
authors.affiliation: Molecular Toxicology and Safety Pharmacology, Global Safety Assessment, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, USA.
authors.forename: Harriet W
authors.initials: HW
authors.lastname: Kamendi
authors.affiliation: Molecular Toxicology, Global Safety Assessment, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, USA.
authors.forename: Clay W
authors.initials: CW
authors.lastname: Scott
authors.affiliation: Molecular Toxicology, Global Safety Assessment, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, USA.
authors.forename: Yvonne P
authors.initials: YP
authors.lastname: Dragan
authors.affiliation: Molecular Toxicology, Global Safety Assessment, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, USA.
authors.forename: Matthew F
authors.initials: MF
authors.lastname: Peters
mesh_terms: D000818:Animals; D002319:Cardiovascular System / Q000187:drug effects*; D004285:Dogs; D004353:Drug Evaluation, Preclinical; D008297:Male; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D047428:Protein Kinase Inhibitors; D020928:Mitogen-Activated Protein Kinases
keywords: cardiotoxicity; impedance; kinase inhibitors.