Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis

Dis Model Mech. 2016 Apr;9(4):389-400. doi: 10.1242/dmm.021790. Epub 2016 Jan 21.

Abstract

The epithelial-to-mesenchymal transition (EMT) is a highly conserved morphogenetic program essential for embryogenesis, regeneration and cancer metastasis. In cancer cells, EMT also triggers cellular reprogramming and chemoresistance, which underlie disease relapse and decreased survival. Hence, identifying compounds that block EMT is essential to prevent or eradicate disseminated tumor cells. Here, we establish a whole-animal-based EMT reporter in zebrafish for rapid drug screening, calledTg(snai1b:GFP), which labels epithelial cells undergoing EMT to producesox10-positive neural crest (NC) cells. Time-lapse and lineage analysis ofTg(snai1b:GFP)embryos reveal that cranial NC cells delaminate from two regions: an early population delaminates adjacent to the neural plate, whereas a later population delaminates from within the dorsal neural tube. TreatingTg(snai1b:GFP)embryos with candidate small-molecule EMT-inhibiting compounds identified TP-0903, a multi-kinase inhibitor that blocked cranial NC cell delamination in both the lateral and medial populations. RNA sequencing (RNA-Seq) analysis and chemical rescue experiments show that TP-0903 acts through stimulating retinoic acid (RA) biosynthesis and RA-dependent transcription. These studies identify TP-0903 as a new therapeutic for activating RAin vivoand raise the possibility that RA-dependent inhibition of EMT contributes to its prior success in eliminating disseminated cancer cells.

Keywords: Drug screen; EMT; Epithelial mesenchymal; Neural crest; Retinoic acid; Zebrafish.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cell Differentiation / drug effects
  • Cell Movement / drug effects
  • Cell Shape / drug effects
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / metabolism
  • Epithelial Cells / cytology*
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Epithelial-Mesenchymal Transition / drug effects*
  • Genes, Reporter*
  • Green Fluorescent Proteins / metabolism
  • Morphogenesis / drug effects*
  • Neural Crest / drug effects
  • Neural Crest / metabolism*
  • Neural Tube / cytology
  • Neural Tube / drug effects
  • Phenotype
  • Pyrimidines / pharmacology
  • Signal Transduction / drug effects
  • Sulfonamides / pharmacology
  • Transcription, Genetic / drug effects
  • Tretinoin / pharmacology*
  • Zebrafish / embryology
  • Zebrafish / metabolism*
  • Zebrafish Proteins / metabolism

Substances

  • Pyrimidines
  • Sulfonamides
  • Zebrafish Proteins
  • Green Fluorescent Proteins
  • dubermatinib
  • Tretinoin