title: Cell-based screen for altered nuclear phenotypes reveals senescence progression in polyploid cells after Aurora kinase B inhibition.
abstract: Cellular senescence is a widespread stress response and is widely considered to be an alternative cancer therapeutic goal. Unlike apoptosis, senescence is composed of a diverse set of subphenotypes, depending on which of its associated effector programs are engaged. Here we establish a simple and sensitive cell-based prosenescence screen with detailed validation assays. We characterize the screen using a focused tool compound kinase inhibitor library. We identify a series of compounds that induce different types of senescence, including a unique phenotype associated with irregularly shaped nuclei and the progressive accumulation of G1 tetraploidy in human diploid fibroblasts. Downstream analyses show that all of the compounds that induce tetraploid senescence inhibit Aurora kinase B (AURKB). AURKB is the catalytic component of the chromosome passenger complex, which is involved in correct chromosome alignment and segregation, the spindle assembly checkpoint, and cytokinesis. Although aberrant mitosis and senescence have been linked, a specific characterization of AURKB in the context of senescence is still required. This proof-of-principle study suggests that our protocol is capable of amplifying tetraploid senescence, which can be observed in only a small population of oncogenic RAS-induced senescence, and provides additional justification for AURKB as a cancer therapeutic target.
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom.
authors.forename: Mahito
authors.initials: M
authors.lastname: Sadaie
authors.affiliation: Cancer Research Technology Discovery Laboratories, Wolfson Institute for Biomedical Research, London WC1E 6BT, United Kingdom.
authors.forename: Christian
authors.initials: C
authors.lastname: Dillon
authors.forename: Masako
authors.initials: M
authors.lastname: Narita
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom Masashi.Narita@cruk.cam.ac.uk.
authors.forename: Masashi
authors.initials: M
authors.lastname: Narita
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom.
authors.forename: Andrew R J
authors.initials: AR
authors.lastname: Young
authors.affiliation: Institute of Cancer Sciences, Wolfson Wohl Cancer Research Centre, University of Glasgow, Glasgow G61 1QH, United Kingdom.
authors.forename: Claire J
authors.initials: CJ
authors.lastname: Cairney
authors.affiliation: St. George's, University of London, London SW17 0RE, United Kingdom.
authors.forename: Lauren S
authors.initials: LS
authors.lastname: Godwin
authors.affiliation: Horizon Discovery Ltd, IQ Cambridge, Waterbeach, Cambridge CB25 9TL, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Torrance
authors.affiliation: St. George's, University of London, London SW17 0RE, United Kingdom.
authors.forename: Dorothy C
authors.initials: DC
authors.lastname: Bennett
authors.affiliation: Institute of Cancer Sciences, Wolfson Wohl Cancer Research Centre, University of Glasgow, Glasgow G61 1QH, United Kingdom.
authors.forename: W Nicol
authors.initials: WN
authors.lastname: Keith
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom.
authors.forename: Masashi
authors.initials: M
authors.lastname: Narita
mesh_terms: D064107:Aurora Kinase B / Q000037:antagonists & inhibitors* / Q000235:genetics; D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D016922:Cellular Senescence / Q000187:drug effects / Q000235:genetics; D020090:Chromosome Segregation; D048749:Cytokinesis / Q000235:genetics; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008938:Mitosis / Q000187:drug effects / Q000235:genetics; D010641:Phenotype; D011123:Polyploidy; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D054852:Small Molecule Libraries / Q000494:pharmacology
chemical_list: D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; D064107:Aurora Kinase B