title: High-Throughput Screening for Readthrough Modulators of CFTR PTC Mutations.
abstract: Cystic fibrosis (CF) is a hereditary disease caused by mutations in the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR). A large number of nearly 2000 reported mutations, including the premature termination codon (PTC) mutations, urgently require new and personalized medicines. We have developed cell-based assays for readthrough modulators of CFTR PTC mutations (or nonsense mutation suppressors), based on the trafficking and surface expression of CFTR. Approximately 85,000 compounds have been screened for two PTC mutations (Y122X and W1282X). The hit rates at the threshold of 50% greater than vehicle response are 2% and 1.4% for CFTR Y122X and CFTR W1282X, respectively. The overlap of the two hit sets at this stringent hit threshold is relatively small. Only ~28% of the hits from the W1282X screen were also hits in the Y122X screen. The overlap increases to ~50% if compounds are included that in the second screen achieve only a less stringent hit criterion, that is, horseradish peroxidase (HRP) activity greater than three standard deviations above the mean of the vehicle. Our data suggest that personalization may not need to address individual genotypes, but that patients with different CFTR PTC mutations could benefit from the same medicines.
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Liang
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Haibo
authors.initials: H
authors.lastname: Shang
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Nikole J
authors.initials: NJ
authors.lastname: Jordan
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Wong
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Dayna
authors.initials: D
authors.lastname: Mercadante
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Josef
authors.initials: J
authors.lastname: Saltz
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Jerome
authors.initials: J
authors.lastname: Mahiou
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Hermann J
authors.initials: HJ
authors.lastname: Bihler
authors.affiliation: 1 CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Mense
mesh_terms: D018389:Codon, Nonsense; D003550:Cystic Fibrosis / Q000188:drug therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D003584:Cytological Techniques / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: C505032:CFTR protein, human; D018389:Codon, Nonsense; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFTR; HTS; PTC; nonsense suppression; readthrough