title: Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis.
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.
authors.affiliation: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Jimenez
authors.affiliation: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Jindong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Monique A
authors.initials: MA
authors.lastname: Morrison
authors.affiliation: Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA.
authors.forename: Clifford
authors.initials: C
authors.lastname: Whatcott
authors.affiliation: Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA.
authors.forename: Katherine K
authors.initials: KK
authors.lastname: Soh
authors.affiliation: Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Warner
authors.affiliation: Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA.
authors.forename: David
authors.initials: D
authors.lastname: Bearss
authors.affiliation: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Cicely A
authors.initials: CA
authors.lastname: Jette
authors.affiliation: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA Rodney.Stewart@hci.utah.edu.
authors.forename: Rodney A
authors.initials: RA
authors.lastname: Stewart
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009024:Morphogenesis / Q000187:drug effects*; D009432:Neural Crest / Q000187:drug effects / Q000378:metabolism*; D054259:Neural Tube / Q000166:cytology / Q000187:drug effects; D010641:Phenotype; D011743:Pyrimidines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013449:Sulfonamides / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology*; D015027:Zebrafish / Q000196:embryology / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000378:metabolism; D000096926:Benzenesulfonamides
chemical_list: D049452:Green Fluorescent Proteins; D011743:Pyrimidines; D013449:Sulfonamides; D014212:Tretinoin; D029961:Zebrafish Proteins; C000606144:dubermatinib; D000096926:Benzenesulfonamides
keywords: Drug screen; EMT; Epithelial mesenchymal; Neural crest; Retinoic acid; Zebrafish