PUBLICATION

Identification of novel osteogenic compounds by an ex-vivo Sp7:luciferase zebrafish scale assay

Authors
de Vrieze, E., Zethof, J., Schulte-Merker, S., Flik, G., Metz, J.R.
ID
ZDB-PUB-150121-7
Date
2015
Source
Bone   74: 106-13 (Journal)
Registered Authors
de Vrieze, Erik, Flik, Gert, Metz, Juriaan R., Schulte-Merker, Stefan
Keywords
Bone, Drug design, Elasmoid scales, Osteoblast, Sp7, Zebrafish
MeSH Terms
  • Animal Structures/metabolism*
  • Animals
  • Animals, Genetically Modified
  • Biological Assay/methods*
  • Bone Morphogenetic Protein 2
  • Luciferases/metabolism*
  • Osteoblasts/metabolism
  • Osteogenesis*
  • Promoter Regions, Genetic/genetics
  • Regeneration
  • Signal Transduction
  • Transcription Factors/genetics
  • Wnt Proteins/metabolism
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
PubMed
25600250 Full text @ Bone
Abstract
Tight interactions among different cell types contributing to bone formation are of key importance in the maintenance of bone homeostasis. Based on the high similarity in responses to (anti)osteogenic signals between zebrafish scales and mammalian bone, we developed and validated a model to screen large numbers of compounds using ex-vivo cultured scales of a Sp7:luciferase transgenic zebrafish. This model combines the high predictive value of explant cultures with quick, sensitive, and quantifiable readout converging the effects via various pathways including WNT-signaling, to SP7/osterix promoter activity. SP7 is pivotal in osteoblast differentiation and activity and its promoter activity provides an excellent surrogate for Sp7 expression. Bmp-2a was shown to dose-dependently increase Sp7-driven luciferase activity ex vivo. Next, we identified novel effects on bone for 51.7% of the compounds from a small library of WNT-signaling modulators, including a strong osteogenic effect for niclosamide. From all previously characterized compounds, the effect on bone was correctly predicted for 70% of compounds, resulting in a 7% false positive- and 21% false negative rate. The proposed Sp7:luciferase zebrafish scale model is unique, powerful and efficient new tool to assess compounds with osteogenic effects, prior to further testing in rodents.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping