title: Novel small molecules potentiate premature termination codon readthrough by aminoglycosides.
abstract: Nonsense mutations introduce premature termination codons and underlie 11% of genetic disease cases. High concentrations of aminoglycosides can restore gene function by eliciting premature termination codon readthrough but with low efficiency. Using a high-throughput screen, we identified compounds that potentiate readthrough by aminoglycosides at multiple nonsense alleles in yeast. Chemical optimization generated phthalimide derivative CDX5-1 with activity in human cells. Alone, CDX5-1 did not induce readthrough or increase TP53 mRNA levels in HDQ-P1 cancer cells with a homozygous TP53 nonsense mutation. However, in combination with aminoglycoside G418, it enhanced readthrough up to 180-fold over G418 alone. The combination also increased readthrough at all three nonsense codons in cancer cells with other TP53 nonsense mutations, as well as in cells from rare genetic disease patients with nonsense mutations in the CLN2, SMARCAL1 and DMD genes. These findings open up the possibility of treating patients across a spectrum of genetic diseases caused by nonsense mutations.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Alireza
authors.initials: A
authors.lastname: Baradaran-Heravi
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Aruna D
authors.initials: AD
authors.lastname: Balgi
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Carla
authors.initials: C
authors.lastname: Zimmerman
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Kunho
authors.initials: K
authors.lastname: Choi
authors.affiliation: The Centre for Drug Research and Development, 2405 Wesbrook Mall, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Fahimeh S
authors.initials: FS
authors.lastname: Shidmoossavee
authors.affiliation: The Centre for Drug Research and Development, 2405 Wesbrook Mall, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Tan
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Célia
authors.initials: C
authors.lastname: Bergeaud
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Krause
authors.affiliation: Department of Zoology and Michael Smith Laboratories, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Flibotte
authors.affiliation: The Centre for Drug Research and Development, 2405 Wesbrook Mall, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Shimizu
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Hilary J
authors.initials: HJ
authors.lastname: Anderson
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, INSERM UMRS974, CNRS FRE3617, Center for Research in Myology, 47 Boulevard de l'hôpital, 75013 Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Mouly
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Eric
authors.initials: E
authors.lastname: Jan
authors.affiliation: The Centre for Drug Research and Development, 2405 Wesbrook Mall, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Tom
authors.initials: T
authors.lastname: Pfeifer
authors.affiliation: The Centre for Drug Research and Development, 2405 Wesbrook Mall, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: James B
authors.initials: JB
authors.lastname: Jaquith
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada michelr@mail.ubc.ca.
authors.forename: Michel
authors.initials: M
authors.lastname: Roberge
mesh_terms: D000483:Alleles; D000617:Aminoglycosides / Q000737:chemistry / Q000494:pharmacology*; D018389:Codon, Nonsense / Q000235:genetics*; D030342:Genetic Diseases, Inborn / Q000235:genetics; D045325:HCT116 Cells; D006720:Homozygote; D006801:Humans; D010303:Paromomycin / Q000494:pharmacology; D010797:Phthalimides / Q000737:chemistry / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013997:Time Factors; D000091346:Tripeptidyl-Peptidase 1; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D000617:Aminoglycosides; D018389:Codon, Nonsense; D010797:Phthalimides; D012333:RNA, Messenger; D054852:Small Molecule Libraries; D000091346:Tripeptidyl-Peptidase 1; D016159:Tumor Suppressor Protein p53; D010303:Paromomycin; C000717267:TPP1 protein, human