title: A high-content small molecule screen identifies sensitivity of glioblastoma stem cells to inhibition of polo-like kinase 1.
abstract: Glioblastoma multiforme (GBM) is the most common primary brain cancer in adults and there are few effective treatments. GBMs contain cells with molecular and cellular characteristics of neural stem cells that drive tumour growth. Here we compare responses of human glioblastoma-derived neural stem (GNS) cells and genetically normal neural stem (NS) cells to a panel of 160 small molecule kinase inhibitors. We used live-cell imaging and high content image analysis tools and identified JNJ-10198409 (J101) as an agent that induces mitotic arrest at prometaphase in GNS cells but not NS cells. Antibody microarrays and kinase profiling suggested that J101 responses are triggered by suppression of the active phosphorylated form of polo-like kinase 1 (Plk1) (phospho T210), with resultant spindle defects and arrest at prometaphase. We found that potent and specific Plk1 inhibitors already in clinical development (BI 2536, BI 6727 and GSK 461364) phenocopied J101 and were selective against GNS cells. Using a porcine brain endothelial cell blood-brain barrier model we also observed that these compounds exhibited greater blood-brain barrier permeability in vitro than J101. Our analysis of mouse mutant NS cells (INK4a/ARF(-/-), or p53(-/-)), as well as the acute genetic deletion of p53 from a conditional p53 floxed NS cell line, suggests that the sensitivity of GNS cells to BI 2536 or J101 may be explained by the lack of a p53-mediated compensatory pathway. Together these data indicate that GBM stem cells are acutely susceptible to proliferative disruption by Plk1 inhibitors and that such agents may have immediate therapeutic value.
authors.affiliation: Samantha Dickson Brain Cancer Unit and Department of Cancer Biology, UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Davide
authors.initials: D
authors.lastname: Danovi
authors.forename: Amos
authors.initials: A
authors.lastname: Folarin
authors.forename: Sabine
authors.initials: S
authors.lastname: Gogolok
authors.forename: Christine
authors.initials: C
authors.lastname: Ender
authors.forename: Ahmed M O
authors.initials: AM
authors.lastname: Elbatsh
authors.forename: Pär G
authors.initials: PG
authors.lastname: Engström
authors.forename: Stefan H
authors.initials: SH
authors.lastname: Stricker
authors.forename: Sladjana
authors.initials: S
authors.lastname: Gagrica
authors.forename: Ana
authors.initials: A
authors.lastname: Georgian
authors.forename: Ding
authors.initials: D
authors.lastname: Yu
authors.forename: Kin Pong
authors.initials: KP
authors.lastname: U
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Harvey
authors.forename: Patrizia
authors.initials: P
authors.lastname: Ferretti
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Paddison
authors.forename: Jane E
authors.initials: JE
authors.lastname: Preston
authors.forename: N Joan
authors.initials: NJ
authors.lastname: Abbott
authors.forename: Paul
authors.initials: P
authors.lastname: Bertone
authors.forename: Austin
authors.initials: A
authors.lastname: Smith
authors.forename: Steven M
authors.initials: SM
authors.lastname: Pollard
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D005909:Glioblastoma / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D006801:Humans; D007189:Indans / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D058953:Neural Stem Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011621:Pteridines / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013552:Swine; D013876:Thiophenes / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency / Q000235:genetics; D000097869:Polo-Like Kinase 1
chemical_list: D001562:Benzimidazoles; D018797:Cell Cycle Proteins; D007189:Indans; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D011621:Pteridines; D011720:Pyrazoles; D054852:Small Molecule Libraries; D013876:Thiophenes; D016159:Tumor Suppressor Protein p53; D000097869:Polo-Like Kinase 1; C505208:(6,7-dimethoxy-2,4-dihydroindeno(1,2-c)pyrazol-3-yl)(3-fluorophenyl)amine; C518477:BI 2536; C541363:BI 6727; C561573:GSK 461364