title: Differential determinants of cancer cell insensitivity to antimitotic drugs discriminated by a one-step cell imaging assay.
abstract: Cancer cells can be drug resistant due to genetic variation at multiple steps in the drug response pathway, including drug efflux pumping, target mutation, and blunted apoptotic response. These are not discriminated by conventional cell survival assays. Here, we report a rapid and convenient high-content cell-imaging assay that measures multiple physiological changes in cells responding to antimitotic small-molecule drugs. Our one-step, no-wash assay uses three dyes to stain living cells and is much more accurate for scoring weakly adherent mitotic and apoptotic cells than conventional antibody-based assays. We profiled responses of 33 cell lines to 8 antimitotic drugs at multiple concentrations and time points using this assay and deposited our data and assay protocols into a public database (http://lincs.hms.harvard.edu/). Our data discriminated between alternative mechanisms that compromise drug sensitivity to paclitaxel and revealed an unexpected bell-shaped dose-response curve for BI2536, a highly selective inhibitor of Polo-like kinases. Our approach can be generalized, is scalable, and should therefore facilitate identification of molecular biomarkers for mechanisms of drug insensitivity in high-throughput screens and other assays.
authors.affiliation: 1Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
authors.forename: Yangzhong
authors.initials: Y
authors.lastname: Tang
authors.forename: Tiao
authors.initials: T
authors.lastname: Xie
authors.forename: Stefan
authors.initials: S
authors.lastname: Florian
authors.forename: Nathan
authors.initials: N
authors.lastname: Moerke
authors.forename: Caroline
authors.initials: C
authors.lastname: Shamu
authors.forename: Cyril
authors.initials: C
authors.lastname: Benes
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Mitchison
mesh_terms: D050256:Antimitotic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000737:chemistry / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D005456:Fluorescent Dyes; D006574:Heterocyclic Compounds, 2-Ring / Q000737:chemistry / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D006801:Humans; D057054:Molecular Imaging / Q000379:methods*; D017239:Paclitaxel / Q000737:chemistry / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011621:Pteridines; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D000097869:Polo-Like Kinase 1
chemical_list: D050256:Antimitotic Agents; D001549:Benzamides; D018797:Cell Cycle Proteins; D005456:Fluorescent Dyes; D006574:Heterocyclic Compounds, 2-Ring; D017239:Paclitaxel; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D011621:Pteridines; D054852:Small Molecule Libraries; D000097869:Polo-Like Kinase 1; C518477:BI 2536
keywords: antimitotic drugs; cancer cells; drug sensitivity; high-content screening; image analysis; live cell imaging assay