Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis
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, ide...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2016-04-01
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| Series: | Disease Models & Mechanisms |
| Subjects: | |
| Online Access: | http://dmm.biologists.org/content/9/4/389 |
| _version_ | 1818047373577814016 |
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| author | Laura Jimenez Jindong Wang Monique A. Morrison Clifford Whatcott Katherine K. Soh Steven Warner David Bearss Cicely A. Jette Rodney A. Stewart |
| author_facet | Laura Jimenez Jindong Wang Monique A. Morrison Clifford Whatcott Katherine K. Soh Steven Warner David Bearss Cicely A. Jette Rodney A. Stewart |
| author_sort | Laura Jimenez |
| collection | DOAJ |
| description | 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, called Tg(snai1b:GFP), which labels epithelial cells undergoing EMT to produce sox10-positive neural crest (NC) cells. Time-lapse and lineage analysis of Tg(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. Treating Tg(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 RA in vivo and raise the possibility that RA-dependent inhibition of EMT contributes to its prior success in eliminating disseminated cancer cells. |
| first_indexed | 2024-12-10T10:04:46Z |
| format | Article |
| id | doaj.art-b5758c079b3648408920bc95ef03755c |
| institution | Directory Open Access Journal |
| issn | 1754-8403 1754-8411 |
| language | English |
| last_indexed | 2024-12-10T10:04:46Z |
| publishDate | 2016-04-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Disease Models & Mechanisms |
| spelling | doaj.art-b5758c079b3648408920bc95ef03755c2022-12-22T01:53:16ZengThe Company of BiologistsDisease Models & Mechanisms1754-84031754-84112016-04-019438940010.1242/dmm.021790021790Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesisLaura Jimenez0Jindong Wang1Monique A. Morrison2Clifford Whatcott3Katherine K. Soh4Steven Warner5David Bearss6Cicely A. Jette7Rodney A. Stewart8 Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA Tolero Pharmaceuticals, Inc., Lehi, UT 84043, USA Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112, USA 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, called Tg(snai1b:GFP), which labels epithelial cells undergoing EMT to produce sox10-positive neural crest (NC) cells. Time-lapse and lineage analysis of Tg(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. Treating Tg(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 RA in vivo and raise the possibility that RA-dependent inhibition of EMT contributes to its prior success in eliminating disseminated cancer cells.http://dmm.biologists.org/content/9/4/389Drug screenEMTEpithelial mesenchymalNeural crestRetinoic acidZebrafish |
| spellingShingle | Laura Jimenez Jindong Wang Monique A. Morrison Clifford Whatcott Katherine K. Soh Steven Warner David Bearss Cicely A. Jette Rodney A. Stewart Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis Disease Models & Mechanisms Drug screen EMT Epithelial mesenchymal Neural crest Retinoic acid Zebrafish |
| title | Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| title_full | Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| title_fullStr | Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| title_full_unstemmed | Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| title_short | Phenotypic chemical screening using a zebrafish neural crest EMT reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| title_sort | phenotypic chemical screening using a zebrafish neural crest emt reporter identifies retinoic acid as an inhibitor of epithelial morphogenesis |
| topic | Drug screen EMT Epithelial mesenchymal Neural crest Retinoic acid Zebrafish |
| url | http://dmm.biologists.org/content/9/4/389 |
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