Oncogenic β-catenin and PIK3CA instruct network states and cancer phenotypes in intestinal organoids

J Cell Biol. 2017 Jun 5;216(6):1567-1577. doi: 10.1083/jcb.201610058. Epub 2017 Apr 25.

Abstract

Colorectal cancer is driven by cooperating oncogenic mutations. In this study, we use organotypic cultures derived from transgenic mice inducibly expressing oncogenic β-catenin and/or PIK3CAH1047R to follow sequential changes in cancer-related signaling networks, intestinal cell metabolism, and physiology in a three-dimensional environment mimicking tissue architecture. Activation of β-catenin alone results in the formation of highly clonogenic cells that are nonmotile and prone to undergo apoptosis. In contrast, coexpression of stabilized β-catenin and PIK3CAH1047R gives rise to intestinal cells that are apoptosis-resistant, proliferative, stem cell-like, and motile. Systematic inhibitor treatments of organoids followed by quantitative phenotyping and phosphoprotein analyses uncover key changes in the signaling network topology of intestinal cells after induction of stabilized β-catenin and PIK3CAH1047R We find that survival and motility of organoid cells are associated with 4EBP1 and AKT phosphorylation, respectively. Our work defines phenotypes, signaling network states, and vulnerabilities of transgenic intestinal organoids as a novel approach to understanding oncogene activities and guiding the development of targeted therapies.

Publication types

  • Video-Audio Media
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Apoptosis
  • Cell Adhesion
  • Cell Movement
  • Cell Proliferation
  • Cell Survival
  • Cell Transformation, Neoplastic / genetics
  • Cell Transformation, Neoplastic / metabolism*
  • Cell Transformation, Neoplastic / pathology
  • Cells, Cultured
  • Class I Phosphatidylinositol 3-Kinases
  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Neoplastic
  • Genetic Predisposition to Disease
  • Humans
  • Intestinal Neoplasms / enzymology*
  • Intestinal Neoplasms / genetics
  • Intestinal Neoplasms / pathology
  • Intestine, Small / enzymology*
  • Intestine, Small / pathology
  • Mice, Transgenic
  • Mutation
  • Neoplastic Stem Cells / enzymology*
  • Neoplastic Stem Cells / pathology
  • Organoids / enzymology*
  • Organoids / pathology
  • Phenotype
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt / metabolism
  • RNA Interference
  • Signal Transduction*
  • Time Factors
  • Transcriptome
  • Transfection
  • beta Catenin / genetics
  • beta Catenin / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • CTNNB1 protein, mouse
  • EIF4EBP1 protein, human
  • Phosphoproteins
  • beta Catenin
  • Phosphatidylinositol 3-Kinases
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • Proto-Oncogene Proteins c-akt