Identification and characterization of a new potent inhibitor targeting CtBP1/BARS in melanoma cells.

Autor: Filograna A; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., De Tito S; Molecular Cell Biology of Autophagy, The Francis Crick Institute, London, UK. The Study Has Been Previously Performed at IEOS-CNR, Naples, Italy., Monte ML; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Oliva R; Department of Chemical Sciences, University of Naples Federico II, 80126, Naples, Italy., Bruzzese F; Animal Facility Unit, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, 80131, Naples, Italy., Roca MS; Experimental Pharmacology Unit, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, Naples, 80131, Italy., Zannetti A; Institute of Biostructures and Bioimaging (IBB), National Research Council (CNR), Naples, 80145, Italy., Greco A; Interdepartmental Service Center of Veterinary Radiology, University of Naples Federico II, 80137, Naples, Italy., Spano D; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Ayala I; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Liberti A; National Research Council (CNR), Piazzale Aldo Moro, 700185, Rome, Italy.; Biology and Evolution of Marine Organisms (BEOM), Stazione Zoologica Anton Dohrn, Naples, Italy., Petraccone L; Department of Chemical Sciences, University of Naples Federico II, 80126, Naples, Italy., Dathan N; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Catara G; Institute of Biochemistry and Cell Biology, National Research Council (CNR), 80131, Naples, Italy., Schembri L; National Research Council (CNR), Piazzale Aldo Moro, 700185, Rome, Italy.; Department of Pharmacy, University of Naples Federico II, 80131, Naples, Italy., Colanzi A; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Budillon A; Scientific Directorate, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, 80131, Naples, Italy., Beccari AR; EXSCALATE, Dompé Farmaceutici S.P.A, Naples, Italy., Del Vecchio P; Department of Chemical Sciences, University of Naples Federico II, 80126, Naples, Italy., Luini A; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy., Corda D; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy. daniela.corda@cnr.it., Valente C; Institute of Experimental Endocrinology and Oncology 'G. Salvatore'(IEOS), National Research Council (CNR), 80131, Naples, Italy. carmen.valente@cnr.it.; Present address: Dompé Farmaceutici S.P.A, L'Aquila, Italy. carmen.valente@cnr.it.
Jazyk: angličtina
Zdroj: Journal of experimental & clinical cancer research : CR [J Exp Clin Cancer Res] 2024 May 06; Vol. 43 (1), pp. 137. Date of Electronic Publication: 2024 May 06.
DOI: 10.1186/s13046-024-03044-5
Abstrakt: Background: The C-terminal-binding protein 1/brefeldin A ADP-ribosylation substrate (CtBP1/BARS) acts both as an oncogenic transcriptional co-repressor and as a fission inducing protein required for membrane trafficking and Golgi complex partitioning during mitosis, hence for mitotic entry. CtBP1/BARS overexpression, in multiple cancers, has pro-tumorigenic functions regulating gene networks associated with "cancer hallmarks" and malignant behavior including: increased cell survival, proliferation, migration/invasion, epithelial-mesenchymal transition (EMT). Structurally, CtBP1/BARS belongs to the hydroxyacid-dehydrogenase family and possesses a NAD(H)-binding Rossmann fold, which, depending on ligands bound, controls the oligomerization of CtBP1/BARS and, in turn, its cellular functions. Here, we proposed to target the CtBP1/BARS Rossmann fold with small molecules as selective inhibitors of mitotic entry and pro-tumoral transcriptional activities.
Methods: Structured-based screening of drug databases at different development stages was applied to discover novel ligands targeting the Rossmann fold. Among these identified ligands, N-(3,4-dichlorophenyl)-4-{[(4-nitrophenyl)carbamoyl]amino}benzenesulfonamide, called Comp.11, was selected for further analysis. Fluorescence spectroscopy, isothermal calorimetry, computational modelling and site-directed mutagenesis were employed to define the binding of Comp.11 to the Rossmann fold. Effects of Comp.11 on the oligomerization state, protein partners binding and pro-tumoral activities were evaluated by size-exclusion chromatography, pull-down, membrane transport and mitotic entry assays, Flow cytometry, quantitative real-time PCR, motility/invasion, and colony assays in A375MM and B16F10 melanoma cell lines. Effects of Comp.11 on tumor growth in vivo were analyzed in mouse tumor model.
Results: We identify Comp.11 as a new, potent and selective inhibitor of CtBP1/BARS (but not CtBP2). Comp.11 directly binds to the CtBP1/BARS Rossmann fold affecting the oligomerization state of the protein (unlike other known CtBPs inhibitors), which, in turn, hinders interactions with relevant partners, resulting in the inhibition of both CtBP1/BARS cellular functions: i) membrane fission, with block of mitotic entry and cellular secretion; and ii) transcriptional pro-tumoral effects with significantly hampered proliferation, EMT, migration/invasion, and colony-forming capabilities. The combination of these effects impairs melanoma tumor growth in mouse models.  CONCLUSIONS: This study identifies a potent and selective inhibitor of CtBP1/BARS active in cellular and melanoma animal models revealing new opportunities to study the role of CtBP1/BARS in tumor biology and to develop novel melanoma treatments.
(© 2024. The Author(s).)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje