title: Identification and characterization of small molecules that inhibit nonsense-mediated RNA decay and suppress nonsense p53 mutations.
abstract: Many of the gene mutations found in genetic disorders, including cancer, result in premature termination codons (PTC) and the rapid degradation of their mRNAs by nonsense-mediated RNA decay (NMD). We used virtual library screening, targeting a pocket in the SMG7 protein, a key component of the NMD mechanism, to identify compounds that disrupt the SMG7-UPF1 complex and inhibit NMD. Several of these compounds upregulated NMD-targeted mRNAs at nanomolar concentrations, with minimal toxicity in cell-based assays. As expected, pharmacologic NMD inhibition disrupted SMG7-UPF1 interactions. When used in cells with PTC-mutated p53, pharmacologic NMD inhibition combined with a PTC "read-through" drug led to restoration of full-length p53 protein, upregulation of p53 downstream transcripts, and cell death. These studies serve as proof-of-concept that pharmacologic NMD inhibitors can restore mRNA integrity in the presence of PTC and can be used as part of a strategy to restore full-length protein in a variety of genetic diseases.
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Leenus
authors.initials: L
authors.lastname: Martin
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Arsen
authors.initials: A
authors.lastname: Grigoryan
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Ding
authors.initials: D
authors.lastname: Wang
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Wang
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Laura
authors.initials: L
authors.lastname: Breda
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Stefano
authors.initials: S
authors.lastname: Rivella
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York.
authors.forename: Timothy
authors.initials: T
authors.lastname: Cardozo
authors.affiliation: Authors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New YorkAuthors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New YorkAuthors' Affiliations: Departments of Medicine, Biochemistry and Molecular Pharmacology; The NYU Cancer Institute, New York University School of Medicine; and Departments of Pediatrics and Cell and Biology Development, Weill Cornell School of Medicine, New York, New York Lawrence.gardner@nyumc.org.
authors.forename: Lawrence B
authors.initials: LB
authors.lastname: Gardner
mesh_terms: D002352:Carrier Proteins / Q000235:genetics; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D018389:Codon, Nonsense; D045325:HCT116 Cells; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D059365:Nonsense Mediated mRNA Decay / Q000187:drug effects* / Q000235:genetics; D020365:RNA Helicases; D012333:RNA, Messenger / Q000235:genetics; D054852:Small Molecule Libraries / Q000494:pharmacology*; D015534:Trans-Activators / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D002352:Carrier Proteins; D018389:Codon, Nonsense; D012333:RNA, Messenger; C494578:SMG7 protein, human; D054852:Small Molecule Libraries; C495901:TP53 protein, human; D015534:Trans-Activators; D016159:Tumor Suppressor Protein p53; D020365:RNA Helicases; C101828:UPF1 protein, human