title: Identification of small molecule inhibitors of the Lin28-mediated blockage of pre-let-7g processing.
abstract: The protein Lin28 and microRNA let-7 play critical roles in mammalian development and human disease. Lin28 inhibits let-7 biogenesis through direct interaction with let-7 precursors (pre-let-7). Accumulating evidence in vitro and in vivo suggests this interaction plays a dominant role in embryonic stem cell self-renewal and tumorigenesis. Thus the Lin28-let-7 interaction might be an attractive drug target, if not for the well-known difficulties in targeting protein-RNA interactions with drugs. The identification and development of suitable probe molecules to further elucidate therapeutic potential, as well as mechanistic details of this pathway will be valuable. We report the development and application of a biophysical high-throughput screening assay for the identification of small molecule inhibitors of the Lin28-pre-let-7 interaction. A library of pharmacologically active small molecules was screened and several small molecule inhibitors were identified and biochemically validated. Of these four validated inhibitors, two compounds successfully restored processing of pre-let-7g in the presence of Lin28, validating the concept. Thus, we have identified examples of small molecule inhibitors of the interaction between Lin28 and pre-let-7. This study provides a proof of concept for small molecule inhibitors that antagonise the effects of Lin28 and enhance processing of let-7 miRNA.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. sb10031@cam.ac.uk.
authors.forename: Helen L
authors.initials: HL
authors.lastname: Lightfoot
authors.affiliation: Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Rd, Cambridge, CB2 1QN, UK. eric.miska@gurdon.cam.ac.uk.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Miska
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. sb10031@cam.ac.uk and Cancer Research UK, Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge, CB2 0RE, UK.
authors.forename: Shankar
authors.initials: S
authors.lastname: Balasubramanian
mesh_terms: D001483:Base Sequence; D004353:Drug Evaluation, Preclinical; D005454:Fluorescence Polarization; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: D035683:MicroRNAs; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D054852:Small Molecule Libraries