PUBLICATION
High-content screen using zebrafish (Danio rerio) embryos identifies a novel kinase activator and inhibitor
- Authors
- Geldenhuys, W.J., Bergeron, S.A., Mullins, J.E., Aljammal, R., Gaasch, B.L., Chen, W.C., Yun, J., Hazlehurst, L.A.
- ID
- ZDB-PUB-170323-23
- Date
- 2017
- Source
- Bioorganic & medicinal chemistry letters 27(9): 2029-2037 (Journal)
- Registered Authors
- Bergeron, Sadie
- Keywords
- Cancer, Compound library, Kinase, Notochord, Phenotypic screen, Somites, Zebrafish
- MeSH Terms
-
- Animals
- Antineoplastic Agents/chemistry
- Antineoplastic Agents/pharmacology
- Benzoic Acid/chemistry
- Benzoic Acid/pharmacology
- Death-Associated Protein Kinases/metabolism
- Drug Discovery/methods*
- Drug Screening Assays, Antitumor/methods
- Embryo, Nonmammalian/drug effects*
- Embryo, Nonmammalian/enzymology
- Enzyme Activation/drug effects
- Enzyme Activators/chemistry*
- Enzyme Activators/pharmacology*
- Neoplasms/drug therapy
- Neoplasms/enzymology
- Protein Kinase Inhibitors/chemistry*
- Protein Kinase Inhibitors/pharmacology*
- Protein Serine-Threonine Kinases/antagonists & inhibitors
- Protein Serine-Threonine Kinases/metabolism
- Zebrafish/embryology*
- Zebrafish Proteins/antagonists & inhibitors
- Zebrafish Proteins/metabolism
- PubMed
- 28320616 Full text @ Bioorg. Med. Chem. Lett.
Citation
Geldenhuys, W.J., Bergeron, S.A., Mullins, J.E., Aljammal, R., Gaasch, B.L., Chen, W.C., Yun, J., Hazlehurst, L.A. (2017) High-content screen using zebrafish (Danio rerio) embryos identifies a novel kinase activator and inhibitor. Bioorganic & medicinal chemistry letters. 27(9):2029-2037.
Abstract
In this report we utilized zebrafish (Danio rerio) embryos in a phenotypical high-content screen (HCS) to identify novel leads in a cancer drug discovery program. We initially validated our HCS model using the flavin adenosine dinucleotide (FAD) containing endoplasmic reticulum (ER) enzyme, endoplasmic reticulum oxidoreductase (ERO1) inhibitor EN460. EN460 showed a dose response effect on the embryos with a dose of 10μM being significantly lethal during early embryonic development. The HCS campaign which employed a small library identified a promising lead compound, a naphthyl-benzoic acid derivative coined compound 1 which had significant dosage and temporally dependent effects on notochord and muscle development in zebrafish embryos. Screening a 369 kinase member panel we show that compound 1 is a PIM3 kinase inhibitor (IC50=4.078μM) and surprisingly a DAPK1 kinase agonist/activator (EC50=39.525μM). To our knowledge this is the first example of a small molecule activating DAPK1 kinase. We provide a putative model for increased phosphate transfer in the ATP binding domain when compound 1 is virtually docked with DAPK1. Our data indicate that observable phenotypical changes can be used in future zebrafish screens to identify compounds acting via similar molecular signaling pathways.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping