title: An epithelial marker promoter induction screen identifies histone deacetylase inhibitors to restore epithelial differentiation and abolishes anchorage independence growth in cancers.
abstract: Epithelial-mesenchymal transition (EMT), a crucial mechanism in development, mediates aggressiveness during carcinoma progression and therapeutic refractoriness. The reversibility of EMT makes it an attractive strategy in designing novel therapeutic approaches. Therefore, drug discovery pipelines for EMT reversal are in need to discover emerging classes of compounds. Here, we outline a pre-clinical drug screening platform for EMT reversal that consists of three phases of drug discovery and validation. From the Phase 1 epithelial marker promoter induction (EpI) screen on a library consisting of compounds being approved by Food and Drug Administration (FDA), Vorinostat (SAHA), a histone deacetylase inhibitor (HDACi), is identified to exert EMT reversal effects by restoring the expression of an epithelial marker, E-cadherin. An expanded screen on 41 HDACi further identifies 28 compounds, such as class I-specific HDACi Mocetinosat, Entinostat and CI994, to restore E-cadherin and ErbB3 expressions in ovarian, pancreatic and bladder carcinoma cells. Mocetinostat is the most potent HDACi to restore epithelial differentiation with the lowest concentration required for 50% induction of epithelial promoter activity (EpIC-50).The HDACi exerts paradoxical effects on EMT transcriptional factors such as SNAI and ZEB family and the effects are context-dependent in epithelial- and mesenchymal-like cells. In vitro functional studies further show that HDACi induced significant increase in anoikis and decrease in spheroid formation in ovarian and bladder carcinoma cells with mesenchymal features. This study demonstrates a robust drug screening pipeline for the discovery of compounds capable of restoring epithelial differentiation that lead to significant functional lethality.
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine , Singapore 117599, Singapore.
authors.forename: H M
authors.initials: HM
authors.lastname: Tang
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine , Singapore 117599, Singapore.
authors.forename: K T
authors.initials: KT
authors.lastname: Kuay
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine , Singapore 117599, Singapore.
authors.forename: P F
authors.initials: PF
authors.lastname: Koh
authors.affiliation: Department of Obstetrics & Gynaecology, National University Health System , Singapore 119228, Singapore.
authors.forename: M
authors.initials: M
authors.lastname: Asad
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine , Singapore 117599, Singapore.
authors.forename: T Z
authors.initials: TZ
authors.lastname: Tan
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine , Singapore 117599, Singapore.
authors.forename: V Y
authors.initials: VY
authors.lastname: Chung
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine, Singapore 117599, Singapore; National University Cancer Institute of Singapore, National University Health System, Singapore 119228, Singapore.
authors.forename: S C
authors.initials: SC
authors.lastname: Lee
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine, Singapore 117599, Singapore; Comprehensive Cancer Center, Institut Gustave Roussy, 114 Rue Edouard Vaillant, 94805 Villejuif, France; CNRS UMR 7057, Matter and Complex Systems, Université Paris Diderot, 5 rue Thomas-Mann, 75013 Paris, France; Department of Biochemistry, National University of Singapore, Singapore 117597, Singapore.
authors.forename: J P
authors.initials: JP
authors.lastname: Thiery
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine NUS Yong Loo Lin School of Medicine, Singapore 117599, Singapore; Department of Obstetrics & Gynaecology, National University Health System, Singapore 119228, Singapore; National University Cancer Institute of Singapore, National University Health System, Singapore 119228, Singapore; Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
authors.forename: Ry-J
authors.initials: RJ
authors.lastname: Huang