Reporters to mark and eliminate basal or luminal epithelial cells in culture and in vivo

PLoS Biol. 2018 Jun 20;16(6):e2004049. doi: 10.1371/journal.pbio.2004049. eCollection 2018 Jun.

Abstract

The contribution of basal and luminal cells to cancer progression and metastasis is poorly understood. We report generation of reporter systems driven by either keratin-14 (K14) or keratin-8 (K8) promoter that not only express a fluorescent protein but also an inducible suicide gene. Transgenic mice express the reporter genes in the right cell compartments of mammary gland epithelia and respond to treatment with toxins. In addition, we engineered the reporters into 4T1 metastatic mouse tumor cell line and demonstrate that K14+ cells, but not K14- or K8+, are both highly invasive in three-dimensional (3D) culture and metastatic in vivo. Treatment of cells in culture, or tumors in mice, with reporter-targeting toxin inhibited both invasive behavior and metastasis in vivo. RNA sequencing (RNA-seq), secretome, and epigenome analysis of K14+ and K14- cells led to the identification of amphoterin-induced protein 2 (Amigo2) as a new cell invasion driver whose expression correlated with decreased relapse-free survival in patients with TP53 wild-type (WT) breast cancer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Division / genetics
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Epithelial Cells / pathology
  • Female
  • Genes, Reporter / genetics*
  • Green Fluorescent Proteins / genetics
  • Keratin-14 / genetics
  • Keratin-8 / genetics
  • Mammary Glands, Animal / cytology
  • Mammary Glands, Animal / pathology*
  • Mammary Neoplasms, Animal / genetics
  • Mammary Neoplasms, Animal / pathology*
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Transgenic
  • Neoplasm Metastasis / pathology
  • Nerve Tissue Proteins / genetics*
  • Nerve Tissue Proteins / metabolism*
  • Promoter Regions, Genetic / genetics

Substances

  • Amigo2 protein, mouse
  • Keratin-14
  • Keratin-8
  • Krt14 protein, mouse
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Green Fluorescent Proteins