title: Zebrafish phenotypic screen identifies novel Notch antagonists.
abstract: Zebrafish represents a powerful in vivo model for phenotype-based drug discovery to identify clinically relevant small molecules. By utilizing this model, we evaluated natural product derived compounds that could potentially modulate Notch signaling that is important in both zebrafish embryogenesis and pathogenic in human cancers. A total of 234 compounds were screened using zebrafish embryos and 3 were identified to be conferring phenotypic alterations similar to embryos treated with known Notch inhibitors. Subsequent secondary screens using HEK293T cells overexpressing truncated Notch1 (HEK293TΔE) identified 2 compounds, EDD3 and 3H4MB, to be potential Notch antagonists. Both compounds reduced protein expression of NOTCH1, Notch intracellular domain (NICD) and hairy and enhancer of split-1 (HES1) in HEK293TΔE and downregulated Notch target genes. Importantly, EDD3 treatment of human oral cancer cell lines demonstrated reduction of Notch target proteins and genes. EDD3 also inhibited proliferation and induced G0/G1cell cycle arrest of ORL-150 cells through inducing p27KIP1. Our data demonstrates the utility of the zebrafish phenotypic screen and identifying EDD3 as a promising Notch antagonist for further development as a novel therapeutic agent.
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Vithya
authors.initials: V
authors.lastname: Velaithan
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Kazuhide Shaun
authors.initials: KS
authors.lastname: Okuda
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Mei Fong
authors.initials: MF
authors.lastname: Ng
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Norazwana
authors.initials: N
authors.lastname: Samat
authors.affiliation: Laboratory of Natural Products, Institute of Bioscience Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.
authors.forename: Sze Wei
authors.initials: SW
authors.lastname: Leong
authors.affiliation: Laboratory of Natural Products, Institute of Bioscience Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.|Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.
authors.forename: Siti Munirah Mohd
authors.initials: SM
authors.lastname: Faudzi
authors.affiliation: Laboratory of Natural Products, Institute of Bioscience Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.|Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.
authors.forename: Faridah
authors.initials: F
authors.lastname: Abas
authors.affiliation: Laboratory of Natural Products, Institute of Bioscience Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.|Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.
authors.forename: Khozirah
authors.initials: K
authors.lastname: Shaari
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Sok Ching
authors.initials: SC
authors.lastname: Cheong
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia.
authors.forename: Pei Jean
authors.initials: PJ
authors.lastname: Tan
authors.affiliation: Cancer Research Malaysia, 12A, Jalan TP5, Taman Perindustrian UEP, 47600, Subang Jaya, Malaysia. vyomesh.patel@cancerresearch.my.
authors.forename: Vyomesh
authors.initials: V
authors.lastname: Patel
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003474:Curcumin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D010641:Phenotype; D051880:Receptors, Notch / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014315:Triterpenes / Q000494:pharmacology*; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D051880:Receptors, Notch; D014315:Triterpenes; D029961:Zebrafish Proteins; D003474:Curcumin
keywords: Human oral cancer; Natural products; Notch signaling; Phenotypic assay; Zebrafish