title: Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures.
abstract: The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from pre-clinical models to clinical trials. Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening. These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm the spatial impact of these dependencies using immunohistochemistry.
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
authors.forename: Eamonn
authors.initials: E
authors.lastname: Morrison
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
authors.forename: Patty
authors.initials: P
authors.lastname: Wai
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
authors.forename: Andri
authors.initials: A
authors.lastname: Leonidou
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
authors.forename: Philip
authors.initials: P
authors.lastname: Bland
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
authors.forename: Saira
authors.initials: S
authors.lastname: Khalique
authors.affiliation: Institute of Cancer Sciences, University of Manchester.
authors.forename: Gillian
authors.initials: G
authors.lastname: Farnie
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research.
authors.forename: Frances
authors.initials: F
authors.lastname: Daley
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research; barrie.peck@icr.ac.uk.
authors.forename: Barrie
authors.initials: B
authors.lastname: Peck
authors.affiliation: The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research; Rachael.Natrajan@icr.ac.uk.
authors.forename: Rachael
authors.initials: R
authors.lastname: Natrajan
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D058534:Class I Phosphatidylinositol 3-Kinases; D023281:Genomics; D006801:Humans; D009369:Neoplasms / Q000235:genetics*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D018874:Spheroids, Cellular
chemical_list: D000970:Antineoplastic Agents; D058534:Class I Phosphatidylinositol 3-Kinases; D019869:Phosphatidylinositol 3-Kinases; D018719:Erb-b2 Receptor Tyrosine Kinases; C484760:PIK3CA protein, human; C508053:ERBB2 protein, human