title: Validation of an in vitro contractility assay using canine ventricular myocytes.
abstract: Measurement of cardiac contractility is a logical part of pre-clinical safety assessment in a drug discovery project, particularly if a risk has been identified or is suspected based on the primary- or non-target pharmacology. However, there are limited validated assays available that can be used to screen several compounds in order to identify and eliminate inotropic liability from a chemical series. We have therefore sought to develop an in vitro model with sufficient throughput for this purpose. Dog ventricular myocytes were isolated using a collagenase perfusion technique and placed in a perfused recording chamber on the stage of a microscope at ~36 °C. Myocytes were stimulated to contract at a pacing frequency of 1 Hz and a digital, cell geometry measurement system (IonOptix™) was used to measure sarcomere shortening in single myocytes. After perfusion with vehicle (0.1% DMSO), concentration-effect curves were constructed for each compound in 4-30 myocytes taken from 1 or 2 dog hearts. The validation test-set was 22 negative and 8 positive inotropes, and 21 inactive compounds, as defined by their effect in dog, cynolomolgous monkey or humans. By comparing the outcome of the assay to the known in vivo contractility effects, the assay sensitivity was 81%, specificity was 75%, and accuracy was 78%. With a throughput of 6-8 compounds/week from 1 cell isolation, this assay may be of value to drug discovery projects to screen for direct contractility effects and, if a hazard is identified, help identify inactive compounds.
authors.affiliation: Safety Assessment UK, AstraZeneca R&D Alderley Park, Macclesfield, SK10 4TG, UK. alex.harmer@astrazeneca.com
authors.forename: A R
authors.initials: AR
authors.lastname: Harmer
authors.forename: N
authors.initials: N
authors.lastname: Abi-Gerges
authors.forename: M J
authors.initials: MJ
authors.lastname: Morton
authors.forename: G F
authors.initials: GF
authors.lastname: Pullen
authors.forename: J P
authors.initials: JP
authors.lastname: Valentin
authors.forename: C E
authors.initials: CE
authors.lastname: Pollard
mesh_terms: D000818:Animals; D004285:Dogs; D055808:Drug Discovery / Q000379:methods; D005260:Female; D006352:Heart Ventricles / Q000187:drug effects; D066298:In Vitro Techniques; D009200:Myocardial Contraction / Q000187:drug effects / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology*; D015203:Reproducibility of Results; D012518:Sarcomeres / Q000502:physiology; D012680:Sensitivity and Specificity; D014741:Video Recording