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Molecules 2018, 23(7), 1691; https://doi.org/10.3390/molecules23071691

A High-Content Screen Reveals New Small-Molecule Enhancers of Ras/Mapk Signaling as Probes for Zebrafish Heart Development

1
Department of Developmental Biology, University of Pittsburgh, BST3, 3501 5th Avenue, Pittsburgh, PA 15213, USA
2
The University of Pittsburgh Drug Discovery Institute, 200 Lothrop Street, Pittsburgh, PA 15260, USA
3
Department of Chemistry, 219 University Drive, University of Pittsburgh, Pittsburgh, PA 15260, USA
4
Department of Cell Biology, University of Pittsburgh, 3500 Terrace Street, Pittsburgh, PA 15213, USA
5
Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA 15213, USA
*
Authors to whom correspondence should be addressed.
Received: 25 June 2018 / Revised: 9 July 2018 / Accepted: 9 July 2018 / Published: 11 July 2018
(This article belongs to the Special Issue Hit Generation and Verification for Novel Lead Compounds)
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Abstract

Zebrafish is the preferred vertebrate model for high throughput chemical screens to discover modulators of complex biological pathways. We adapted a transgenic zebrafish line, Tg(dusp6:EGFP), which reports on fibroblast growth factor (Fgf)/Ras/Mapk activity, into a quantitative, high-content chemical screen to identify novel Fgf hyperactivators as chemical probes for zebrafish heart development and regeneration. We screened 10,000 compounds from the TimTec ActiProbe library, and identified several structurally distinct classes of molecules that enhanced Fgf/Ras/Mapk signaling. We chose three agents—ST020101, ST011282, and ST006994—for confirmatory and functional studies based on potency, repeatability with repurchased material, favorable whole organism toxicity, and evidence of structure–activity relationships. Functional follow-up assays confirmed that all three compounds induced the expression of Fgf target genes during zebrafish embryonic development. Moreover, these compounds increased cardiac progenitor populations by effecting a fate change from endothelial to cardiac progenitors that translated into increased numbers of cardiomyocytes. Interestingly, ST006994 augmented Fgf/Ras/Mapk signaling without increasing Erk phosphorylation, suggesting a molecular mechanism of action downstream of Erk. We posit that the ST006994 pharmacophore could become a unique chemical probe to uncover novel mechanisms of Fgf signaling during heart development and regeneration downstream of the Mapk signaling node. View Full-Text
Keywords: zebrafish; high-throughput screening; high-content analysis; Cognition Network Technology; Fgf signaling; heart development; probe discovery zebrafish; high-throughput screening; high-content analysis; Cognition Network Technology; Fgf signaling; heart development; probe discovery
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Saydmohammed, M.; Vollmer, L.L.; Onuoha, E.O.; Maskrey, T.S.; Gibson, G.; Watkins, S.C.; Wipf, P.; Vogt, A.; Tsang, M. A High-Content Screen Reveals New Small-Molecule Enhancers of Ras/Mapk Signaling as Probes for Zebrafish Heart Development. Molecules 2018, 23, 1691.

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