title: A 1536-well quantitative high-throughput screen to identify compounds targeting cancer stem cells.
abstract: Tumor cell subpopulations called cancer stem cells (CSCs) or tumor-initiating cells (TICs) have self-renewal potential and are thought to drive metastasis and tumor formation. Data suggest that these cells are resistant to current chemotherapy and radiation therapy treatments, leading to cancer recurrence. Therefore, finding new drugs and/or drug combinations that cause death of both the differentiated tumor cells as well as CSC populations is a critical unmet medical need. Here, we describe how cancer-derived CSCs are generated from cancer cell lines using stem cell growth media and nonadherent conditions in quantities that enable high-throughput screening (HTS). A cell growth assay in a 1536-well microplate format was developed with these CSCs and used to screen a focused collection of oncology drugs and clinical candidates to find compounds that are cytotoxic against these highly aggressive cells. A hit selection process that included potency and efficacy measurements during the primary screen allowed us to efficiently identify compounds with potent cytotoxic effects against spheroid-derived CSCs. Overall, this research demonstrates one of the first miniaturized HTS assays using CSCs. The procedures described here should enable further testing of the effect of compounds on CSCs and help determine which pathways need to be targeted to kill them.
authors.affiliation: Division of Preclinical Innovation, National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, MD 20850, USA. mathewsla@mail.nih.gov
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Mathews
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Keller
authors.forename: Bonnie L
authors.initials: BL
authors.lastname: Goodwin
authors.forename: Rajarshi
authors.initials: R
authors.lastname: Guha
authors.forename: Paul
authors.initials: P
authors.lastname: Shinn
authors.forename: Rebecca
authors.initials: R
authors.lastname: Mull
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Thomas
authors.forename: Rachel L
authors.initials: RL
authors.lastname: de Kluyver
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Sayers
authors.forename: Marc
authors.initials: M
authors.lastname: Ferrer
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D000071317:Nanog Homeobox Protein; D009369:Neoplasms / Q000188:drug therapy; D014411:Neoplastic Stem Cells / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D054852:Small Molecule Libraries