PUBLICATION

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

Authors
Saydmohammed, M., Vollmer, L.L., Onuoha, E.O., Maskrey, T.S., Gibson, G., Watkins, S.C., Wipf, P., Vogt, A., Tsang, M.
ID
ZDB-PUB-180713-14
Date
2018
Source
Molecules   23(7): (Journal)
Registered Authors
Tsang, Michael
Keywords
Cognition Network Technology, Fgf signaling, heart development, high-content analysis, high-throughput screening, probe discovery, zebrafish
MeSH Terms
  • Molecular Probes/chemistry*
  • Zebrafish/embryology*
  • Animals
  • ras Proteins/metabolism*
  • Fibroblast Growth Factors/metabolism
  • High-Throughput Screening Assays/methods*
  • Small Molecule Libraries/chemistry
  • Small Molecule Libraries/pharmacology*
  • MAP Kinase Signaling System*/drug effects
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/metabolism
  • Extracellular Signal-Regulated MAP Kinases/metabolism
  • Heart/drug effects
  • Heart/embryology*
  • Organ Size/drug effects
(all 15)
PubMed
29997348 Full text @ Molecules
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.
Genes / Markers
Figures
Figure Gallery (3 images)
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Expression
No data available
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
f2TgTransgenic Insertion
    pt6TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      No data available
      Fish
      No data available
      Antibodies
      Name Type Antigen Genes Isotypes Host Organism
      Ab2-mapkmonoclonalIgG1Mouse
      Ab11-mapkmonoclonal
        IgGRabbit
        1 - 2 of 2
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        Orthology
        No data available
        Engineered Foreign Genes
        Marker Marker Type Name
        d2EGFPEFGd2EGFP
        DsRed2EFGDsRed2
        1 - 2 of 2
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        Mapping
        No data available