title: Targeting the DNA-binding activity of the human ERG transcription factor using new heterocyclic dithiophene diamidines.
abstract: Direct modulation of gene expression by targeting oncogenic transcription factors is a new area of research for cancer treatment. ERG, an ETS-family transcription factor, is commonly over-expressed or translocated in leukaemia and prostate carcinoma. In this work, we selected the di-(thiophene-phenyl-amidine) compound DB1255 as an ERG/DNA binding inhibitor using a screening test of synthetic inhibitors of the ERG/DNA interaction followed by electrophoretic mobility shift assays (EMSA) validation. Spectrometry, footprint and biosensor-surface plasmon resonance analyses of the DB1255/DNA interaction evidenced sequence selectivity and groove binding as dimer. Additional EMSA evidenced the precise DNA-binding sequence required for optimal DB1255/DNA binding and thus for an efficient ERG/DNA complex inhibition. We further highlighted the structure activity relationships from comparison with derivatives. In cellulo luciferase assay confirmed this modulation both with the constructed optimal sequences and the Osteopontin promoter known to be regulated by ERG and which ERG-binding site was protected from DNaseI digestion on binding of DB1255. These data showed for the first time the ERG/DNA complex modulation, both in vitro and in cells, by a heterocyclic diamidine that specifically targets a portion of the ERG DNA recognition site.
authors.affiliation: INSERM UMR837-JPARC, Team 4, Molecular and Cellular Targeting for Cancer Treatment, University of Lille North of France, IMPRT-IFR114, Institut pour la Recherche sur le Cancer de Lille, Place de Verdun, Lille F-59045, France.
authors.forename: Raja
authors.initials: R
authors.lastname: Nhili
authors.forename: Paul
authors.initials: P
authors.lastname: Peixoto
authors.forename: Sabine
authors.initials: S
authors.lastname: Depauw
authors.forename: Sébastien
authors.initials: S
authors.lastname: Flajollet
authors.forename: Xavier
authors.initials: X
authors.lastname: Dezitter
authors.forename: Manoj M
authors.initials: MM
authors.lastname: Munde
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Ismail
authors.forename: Arvind
authors.initials: A
authors.lastname: Kumar
authors.forename: Abdelbasset A
authors.initials: AA
authors.lastname: Farahat
authors.forename: Chad E
authors.initials: CE
authors.lastname: Stephens
authors.forename: Martine
authors.initials: M
authors.lastname: Duterque-Coquillaud
authors.forename: W
authors.initials: W
authors.lastname: David Wilson
authors.forename: David W
authors.initials: DW
authors.lastname: Boykin
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: David-Cordonnier
mesh_terms: D000578:Amidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001665:Binding Sites; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D006801:Humans; D013876:Thiophenes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015534:Trans-Activators / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*; D000071230:Transcriptional Regulator ERG
chemical_list: D000578:Amidines; D000970:Antineoplastic Agents; C579039:DB1255; C072312:ERG protein, human; D013876:Thiophenes; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG; D004247:DNA