title: PIK3CA mutations and TP53 alterations cooperate to increase cancerous phenotypes and tumor heterogeneity.
abstract: The combined contributions of oncogenes and tumor suppressor genes toward carcinogenesis remain poorly understood. Elucidation of cancer gene cooperativity can provide new insights leading to more effective use of therapies.
  
  
  
  We used somatic cell genome editing to introduce singly and in combination PIK3CA mutations (E545K or H1047R) with TP53 alterations (R248W or knockout), to assess any enhanced cancerous phenotypes. The non-tumorigenic human breast epithelial cell line, MCF10A, was used as the parental cell line, and resultant cells were assessed via various in vitro assays, growth as xenografts, and drug sensitivity assays using targeted agents and chemotherapies.
  
  
  
  Compared to single-gene-targeted cells and parental controls, cells with both a PIK3CA mutation and TP53 alteration had increased cancerous phenotypes including cell proliferation, soft agar colony formation, aberrant morphology in acinar formation assays, and genomic heterogeneity. Cells also displayed varying sensitivities to anti-neoplastic drugs, although all cells with PIK3CA mutations showed a relative increased sensitivity to paclitaxel. All cell lines remained non-tumorigenic.
  
  
  
  This cell line panel provides a resource for further elucidating cooperative genetic mediators of carcinogenesis and response to therapies.
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Croessmann
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Hong Yuen
authors.initials: HY
authors.lastname: Wong
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Zabransky
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: David
authors.initials: D
authors.lastname: Chu
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: D Marc
authors.initials: DM
authors.lastname: Rosen
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.|Oncology iMED, AstraZeneca, 35 Gatehouse Dr., Waltham, MA, 02451, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Cidado
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Rory L
authors.initials: RL
authors.lastname: Cochran
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: W Brian
authors.initials: WB
authors.lastname: Dalton
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Bracha
authors.initials: B
authors.lastname: Erlanger
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Cravero
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Berry
authors.initials: B
authors.lastname: Button
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Kyker-Snowman
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Paula J
authors.initials: PJ
authors.lastname: Hurley
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA.
authors.forename: Josh
authors.initials: J
authors.lastname: Lauring
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Room 151, Baltimore, MD, 21287, USA. bpark2@jhmi.edu.|Department of Chemical and Biomolecular Engineering, The Whiting School of Engineering, The Johns Hopkins University, Baltimore, MD, 21218, USA. bpark2@jhmi.edu.
authors.forename: Ben Ho
authors.initials: BH
authors.lastname: Park
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002503:Centromere / Q000235:genetics; D058534:Class I Phosphatidylinositol 3-Kinases / Q000235:genetics*; D056915:DNA Copy Number Variations; D004195:Disease Models, Animal; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D000072669:Gene Editing; D055786:Gene Knockout Techniques; D042822:Genomic Instability; D005838:Genotype; D006801:Humans; D051379:Mice; D009154:Mutation; D017239:Paclitaxel / Q000494:pharmacology; D010641:Phenotype; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D016159:Tumor Suppressor Protein p53; D058534:Class I Phosphatidylinositol 3-Kinases; C484760:PIK3CA protein, human; D017239:Paclitaxel
keywords: Breast cancer; PIK3CA; TP53; Tumor heterogeneity