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Cited by 134Cystic Fibrosis Research AdvancesGenetic and Kidney Cyst DiseasesIon Transport and Channel Regulation

Structure-guided combination therapy to potently improve the function of mutant CFTRs

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Veit, Guido, Xu, Haijin, Dreano, Elise, Avramescu, Radu G., Bagdany, Miklos, Beitel, Lenore K., Roldan, Ariel, Hancock, Mark A. +14 more·2018·Nature Medicine
DOI 10.1038/s41591-018-0200-xPMC PMC6301090
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Abstract

Available corrector drugs are unable to effectively rescue the folding defects of CFTR-ΔF508 (or CFTR-F508del), the most common disease-causing mutation of the cystic fibrosis transmembrane conductance regulator, a plasma membrane (PM) anion channel, and thus to substantially ameliorate clinical phenotypes of cystic fibrosis (CF). To overcome the corrector efficacy ceiling, here we show that compounds targeting distinct structural defects of CFTR can synergistically rescue mutant expression and function at the PM. High-throughput cell-based screens and mechanistic analysis identified three small-molecule series that target defects at nucleotide-binding domain (NBD1), NBD2 and their membrane-spanning domain (MSD) interfaces. Although individually these compounds marginally improve ΔF508-CFTR folding efficiency, function and stability, their combinations lead to ~50-100% of wild-type-level correction in immortalized and primary human airway epithelia and in mouse nasal epithelia. Likewise, corrector combinations were effective against rare missense mutations in various CFTR domains, probably acting via structural allostery, suggesting a mechanistic framework for their broad application.

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  • PM density values were normalized with cell viability measured by alamarBlue Assay and related to WT-CFTR by normalizing with CFTR mRNA abundance determined by qPCR.

    “PM density values were normalized with cell viability measured by alamarBlue Assay (Invitrogen) and related to WT-CFTR by normalizing with CFTR mRNA abundance determined by qPCR as previously (21).”

    From Allosteric folding correction of F508del and rare CFTR mutants by elexacaftor-tezacaftor-ivacaftor (Trikafta) combination (2020) · cites this paper as [21]

    The Online Methods section “PM density measurement” (page unavailable) explicitly says PM density was normalized to cell viability measured by the alamarBlue Assay. The “qPCR” section describes measuring CFTR transcript abundance, but our copy does not say that PM density was normalized by that abundance to relate it to WT-CFTR. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • Chronic treatment with 1 μM VX-770 significantly decreased functional correction by VX-809 in the presence of acute potentiator addition.

    “Chronic treatment with 1 μM VX-770, a concentration that is therapeutically reached in respiratory epithelia (34), was reported to significantly decrease the functional correction by VX-809 in the presence of acute potentiator addition (Figure 3, A and B), as observed in CF-HBE (35, 36) and CF-HNE (21).”

    From Allosteric folding correction of F508del and rare CFTR mutants by elexacaftor-tezacaftor-ivacaftor (Trikafta) combination (2020) · cites this paper as [21]

    The Results section on functional correction in bronchial and nasal epithelia (page unavailable) states that chronic VX-770 treatment decreased VX-809 corrector efficacy in CF-HNE, consistent with the reported CF-HBE response. The passages do not establish the specified 1 μM concentration, statistical significance, or that the decrease was measured in the presence of acute potentiator addition. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • VX-809 treatment increased forskolin-activated and acutely VX-770-potentiated Isc of F508del-CFTR from 9% to 25% relative to WT.

    “VX-809 treatment increased the forskolin-activated and acutely VX-770–potentiated short-circuit current (Isc) of F508del-CFTR from 9% to 25%, relative to that of the WT (Figure 3, A and B, and Supplemental Figure 3) as reported before (21, 33).”

    From Allosteric folding correction of F508del and rare CFTR mutants by elexacaftor-tezacaftor-ivacaftor (Trikafta) combination (2020) · cites this paper as [21]

    The Results section, “Functional correction of human CFTRΔF508/ΔF508 bronchial and nasal epithelia” (page unavailable), confirms that VX-809 increased forskolin-activated, potentiated F508del-CFTR short-circuit current relative to WT. It reports an increase from 6% to 27%, however, not the claimed 9% to 25%; the 9% figure appears in a separate result for forskolin-activated current. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Combination of VX-445 with the type II corrector 3151 increased F508del-CFTR PM density to ~25% of WT.

    “Combination of VX-445 with the investigational type II corrector 3151 (21) also increased the F508del-CFTR PM density to ~25% of the WT (Figure 1B).”

    From Allosteric folding correction of F508del and rare CFTR mutants by elexacaftor-tezacaftor-ivacaftor (Trikafta) combination (2020) · cites this paper as [21]

    The Results (Clustering corrector hits based on mechanism of action; page unavailable) identify 3151 as a type II corrector. The Results (Cooperative rescue of biogenesis, PM expression and stability by corrector combinations; page unavailable) report increased F508del-CFTR PM density with combinations containing 3151, but the combinations described use VX-809 and 4172, not VX-445. Our copy does not establish the claimed VX-445–3151 result or its ~25%-of-WT value. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • VX-661 plus VX-445 permits allosteric correction of the global conformation defect elicited by other missense CF mutation located in the MSD1, MSD2, NBD1, or NBD2.

    “The VX-661 plus VX-445 also permits allosteric correction of the global conformation defect elicited by other missense CF mutation located in the MSD1, MSD2, NBD1, or NBD2, in agreement with our previous results (21).”

    From Allosteric folding correction of F508del and rare CFTR mutants by elexacaftor-tezacaftor-ivacaftor (Trikafta) combination (2020) · cites this paper as [21]

    The Discussion (page unavailable) says that corrector combinations can allosterically correct conformational defects caused by missense mutations in MSD1, MSD2, NBD1, or NBD2. The Results identify the combination tested for these mutants as VX-809, 3151, and 4172, not VX-661 plus VX-445. The cited paper therefore supports the allosteric-correction principle but not its attribution to that specific drug pair. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Cites

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Cited by

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Authors
Veit, G., Xu, H., Dreano, E., Avramescu, R. G., Bagdany, M., Beitel, L. K., Roldan, A., Hancock, M. A., Lay, C., Li, W., Morin, K., Gao, S., Mak, P. A., Ainscow, E., Orth, A. P., McNamara, P., Edelman, A., Frenkiel, S., Matouk, E., Sermet-Gaudelus, I., Barnes, W. G., & Lukacs, G. L.
Affiliations
  • McGill University (CA)
  • Genomics Institute of the Novartis Research Foundation (US)
  • Inserm (FR)
  • Montreal Chest Institute (CA)
Corresponding author
Veit, Guido
ORCID
Venue
Nature Medicine
Publisher
Springer Science and Business Media LLC
Published
2018-10-08
Volume
24
Issue
11
Pages
1732-1742
ISSN
  • 1078-8956 (ISSN-L)
  • 1546-170X
DOI
10.1038/s41591-018-0200-x
PMC
PMC6301090
OpenAlex
W2893878927
Cited by
134
References
1
Citations by year
2026: 5 · 2025: 11 · 2024: 13 · 2023: 8 · 2022: 25 · 2021: 30 · 2020: 24 · 2019: 16 · 2018: 1 · 2017: 1
FWCI
13.37
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK075302)
Topics
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
Keywords
CFTR, cystic fibrosis, combination therapy, protein folding defects, nucleotide-binding domain, transmembrane domain, small molecules, missense mutations, airway epithelium
MeSH terms
Protein Domains, Allosteric Regulation, Bronchi, Cell Membrane, Cystic Fibrosis, Epithelial Cells, Gene Expression Regulation, Humans, Mutation, Nasal Mucosa, Structure-Activity Relationship, Protein Folding, Cystic Fibrosis Transmembrane Conductance Regulator, Small Molecule Libraries

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