title: A molecular screening approach to identify and characterize inhibitors of glioblastoma stem cells.
abstract: Glioblastoma (GBM) is among the most lethal of all cancers. GBM consist of a heterogeneous population of tumor cells among which a tumor-initiating and treatment-resistant subpopulation, here termed GBM stem cells, have been identified as primary therapeutic targets. Here, we describe a high-throughput small molecule screening approach that enables the identification and characterization of chemical compounds that are effective against GBM stem cells. The paradigm uses a tissue culture model to enrich for GBM stem cells derived from human GBM resections and combines a phenotype-based screen with gene target-specific screens for compound identification. We used 31,624 small molecules from 7 chemical libraries that we characterized and ranked based on their effect on a panel of GBM stem cell-enriched cultures and their effect on the expression of a module of genes whose expression negatively correlates with clinical outcome: MELK, ASPM, TOP2A, and FOXM1b. Of the 11 compounds meeting criteria for exerting differential effects across cell types used, 4 compounds showed selectivity by inhibiting multiple GBM stem cells-enriched cultures compared with nonenriched cultures: emetine, n-arachidonoyl dopamine, n-oleoyldopamine (OLDA), and n-palmitoyl dopamine. ChemBridge compounds #5560509 and #5256360 inhibited the expression of the 4 mitotic module genes. OLDA, emetine, and compounds #5560509 and #5256360 were chosen for more detailed study and inhibited GBM stem cells in self-renewal assays in vitro and in a xenograft model in vivo. These studies show that our screening strategy provides potential candidates and a blueprint for lead compound identification in larger scale screens or screens involving other cancer types.
authors.affiliation: Intellectual and Developmental Disabilities Research Center, Department of Psychiatry, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA.
authors.forename: Koppany
authors.initials: K
authors.lastname: Visnyei
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Onodera
authors.forename: Robert
authors.initials: R
authors.lastname: Damoiseaux
authors.forename: Kuniyasu
authors.initials: K
authors.lastname: Saigusa
authors.forename: Syuzanna
authors.initials: S
authors.lastname: Petrosyan
authors.forename: David
authors.initials: D
authors.lastname: De Vries
authors.forename: Denise
authors.initials: D
authors.lastname: Ferrari
authors.forename: Jonathan
authors.initials: J
authors.lastname: Saxe
authors.forename: Eduard H
authors.initials: EH
authors.lastname: Panosyan
authors.forename: Michael
authors.initials: M
authors.lastname: Masterman-Smith
authors.forename: Jack
authors.initials: J
authors.lastname: Mottahedeh
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Bradley
authors.forename: Jing
authors.initials: J
authors.lastname: Huang
authors.forename: Chiara
authors.initials: C
authors.lastname: Sabatti
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakano
authors.forename: Harley I
authors.initials: HI
authors.lastname: Kornblum
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000378:metabolism / Q000473:pathology; D016895:Culture Media, Serum-Free; D004640:Emetine / Q000494:pharmacology; D004815:Epidermal Growth Factor; D005346:Fibroblast Growth Factors; D015870:Gene Expression / Q000187:drug effects; D005909:Glioblastoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D016895:Culture Media, Serum-Free; D005346:Fibroblast Growth Factors; D004815:Epidermal Growth Factor; D004640:Emetine