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Discovery of novel potent ?F508-CFTR correctors that target the nucleotide binding domain.

ABSTRACT: The deletion of Phe508 (?F508) in the first nucleotide binding domain (NBD1) of CFTR is the most common mutation associated with cystic fibrosis. The ?F508-CFTR mutant is recognized as improperly folded and targeted for proteasomal degradation. Based on molecular dynamics simulation results, we hypothesized that interaction between ?F508-NBD1 and housekeeping proteins prevents ?F508-CFTR delivery to the plasma membrane. Based on this assumption we applied structure-based virtual screening to identify new low-molecular-weight compounds that should bind to ?F508-NBD1 and act as protein-protein interaction inhibitors. Using different functional assays for CFTR activity, we demonstrated that in silico-selected compounds induced functional expression of ?F508-CFTR in transfected HeLa cells, human bronchial CF cells in primary culture, and in the nasal epithelium of homozygous ?F508-CFTR mice. The proposed compounds disrupt keratin8-?F508-CFTR interaction in ?F508-CFTR HeLa cells. Structural analysis of ?F508-NBD1 in the presence of these compounds suggests their binding to NBD1. We conclude that our strategy leads to the discovery of new compounds that are among the most potent correctors of ?F508-CFTR trafficking defect known to date.

SUBMITTER: Odolczyk N

PROVIDER: S-EPMC3799575 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature

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Discovery of novel potent ΔF508-CFTR correctors that target the nucleotide binding domain.

Odolczyk Norbert N Fritsch Janine J Norez Caroline C Servel Nathalie N da Cunha Melanie Faria MF Bitam Sara S Kupniewska Anna A Wiszniewski Ludovic L Colas Julien J Tarnowski Krzysztof K Tondelier Danielle D Roldan Ariel A Saussereau Emilie L EL Melin-Heschel Patricia P Wieczorek Grzegorz G Lukacs Gergely L GL Dadlez Michal M Faure Grazyna G Herrmann Harald H Ollero Mario M Becq Frédéric F Zielenkiewicz Piotr P Edelman Aleksander A

EMBO molecular medicine 20130827 10

The deletion of Phe508 (ΔF508) in the first nucleotide binding domain (NBD1) of CFTR is the most common mutation associated with cystic fibrosis. The ΔF508-CFTR mutant is recognized as improperly folded and targeted for proteasomal degradation. Based on molecular dynamics simulation results, we hypothesized that interaction between ΔF508-NBD1 and housekeeping proteins prevents ΔF508-CFTR delivery to the plasma membrane. Based on this assumption we applied structure-based virtual screening to ide ...[more]

PMID: 23982976

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Project description:Premature-termination codons (PTCs) in CFTR (cystic fibrosis [CF] transmembrane conductance regulator) result in nonfunctional CFTR protein and are the proximate cause of ∼11% of CF-causing alleles, for which no treatments exist. The CFTR corrector lumacaftor and the potentiator ivacaftor improve CFTR function with terminal PTC mutations and enhance the effect of readthrough agents. Novel correctors GLPG2222 (corrector 1 [C1]), GLPG3221 (corrector 2 [C2]), and potentiator GLPG1837 compare favorably with lumacaftor and ivacaftor in vitro. Here, we evaluated the effect of correctors C1a and C2a (derivatives of C1 and C2) and GLPG1837 alone or in combination with the readthrough compound G418 on CFTR function using heterologous Fischer rat thyroid (FRT) cells, the genetically engineered human bronchial epithelial (HBE) 16HBE14o- cell lines, and primary human cells with PTC mutations. In FRT lines pretreated with G418, GLPG1837 elicited dose-dependent increases in CFTR activity that exceeded those from ivacaftor in FRT-W1282X and FRT-R1162X cells. A three-mechanism strategy consisting of G418, GLPG1837, and two correctors (C1a + C2a) yielded the greatest functional improvements in FRT and 16HBE14o- PTC variants, noting that correction and potentiation without readthrough was sufficient to stimulate CFTR activity for W1282X cells. GLPG1837 + C1a + C2a restored substantial function in G542X/F508del HBE cells and restored even more function for W1282X/F508del cells, largely because of the corrector/potentiator effect, with no additional benefit from G418. In G542X/R553X or R1162X/R1162X organoids, enhanced forskolin-induced swelling was observed with G418 + GLPG1837 + C1a + C2a, although GLPG1837 + C1a + C2a alone was sufficient to improve forskolin-induced swelling in W1282X/W1282X organoids. Combination of CFTR correctors, potentiators, and readthrough compounds augments the functional repair of CFTR nonsense mutations, indicating the potential for novel correctors and potentiators to restore function to truncated W1282X CFTR.

Dataset Information

Discovery of novel potent ?F508-CFTR correctors that target the nucleotide binding domain.

Publications

Discovery of novel potent ΔF508-CFTR correctors that target the nucleotide binding domain.

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