title: The GSK3β inhibitor BIS I reverts YAP-dependent EMT signature in PDAC cell lines by decreasing SMADs expression level.
abstract: The Yes-associated protein, YAP, is a transcriptional co-activator, mediating the Epithelial to Mesenchymal Transition program in pancreatic ductal adenocarcinoma (PDAC). With the aim to identify compounds that can specifically modulate YAP functionality in PDAC cell lines, we performed a small scale, drug-based screening experiment using YAP cell localization as the read-out. We identified erlotinib as an inducer of YAP cytoplasmic localization, an inhibitor of the TEA luciferase reporter system and the expression of the bona fide YAP target gene, Connective Tissue Growth Factor CTGF. On the other hand, BIS I, an inhibitor of PKCδ and GSK3β, caused YAP accumulation into the nucleus. Activation of β-catenin reporter and interfering experiments show that inhibition of the PKCδ/GSK3β pathway triggers YAP nuclear accumulation inducing YAP/TEAD transcriptional response. Inhibition of GSK3β by BIS I reduced the expression levels of SMADs protein and reduced YAP contribution to EMT. Notably, BIS I reduced proliferation, migration and clonogenicity of PDAC cells in vitro, phenocopying YAP genetic down-regulation. As shown by chromatin immunoprecipitation experiments and YAP over-expressing rescue experiments, BIS I reverted YAP-dependent EMT program by modulating the expression of the YAP target genes E-cadherin, vimentin, CTGF and of the newly identified target, CD133. In conclusion, we identified two different molecules, erlotinib and BIS I, modulating YAP functionality although via different mechanisms of action, with the second one specifically inhibiting the YAP-dependent EMT program in PDAC cell lines.
authors.affiliation: Laboratory of Genomic Screening, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Natthakan
authors.initials: N
authors.lastname: Thongon
authors.affiliation: Laboratory of Gene Expression and Muscular Dystrophy, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Castiglioni
authors.affiliation: Laboratory of Genomic Screening, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Zucal
authors.affiliation: Laboratory of Genomic Screening, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Latorre
authors.affiliation: Laboratory of Genomic Screening, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Vito
authors.initials: V
authors.lastname: D'Agostino
authors.affiliation: Department of Internal Medicine, University of Genoa, Genoa, Italy.
authors.forename: Inga
authors.initials: I
authors.lastname: Bauer
authors.affiliation: High Throughput Screening Facility, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Michael
authors.initials: M
authors.lastname: Pancher
authors.affiliation: Department of Internal Medicine, University of Genoa, Genoa, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ballestrero
authors.affiliation: Laboratory of Pancreatic Cancer Translational Research, Clinic University of Bonn, Bonn, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Feldmann
authors.affiliation: Department of Internal Medicine, University of Genoa, Genoa, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Nencioni
authors.affiliation: Laboratory of Genomic Screening, Centre for Integrative Biology, University of Trento, Trento, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Provenzani
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D021441:Carcinoma, Pancreatic Ductal / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D000069347:Erlotinib Hydrochloride / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D008301:Maleimides / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D010750:Phosphoproteins / Q000378:metabolism; D051785:Smad Proteins / Q000096:biosynthesis; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; D007211:Indoles; D008301:Maleimides; D010750:Phosphoproteins; D051785:Smad Proteins; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; D000069347:Erlotinib Hydrochloride; D000071679:Glycogen Synthase Kinase 3 beta; C070515:bisindolylmaleimide I
keywords: CTGF; EMT; PDAC; YAP; bisindolylmaleimides