Additive Potentiation of R334W-CFTR Function by Novel Small Molecules
The R334W (c.1000C>T, p.Arg334Trp) is a rare cystic fibrosis (CF)-causing mutation for which no causal therapy is currently approved. This mutation leads to a significant reduction of CF transmembrane conductance regulator (CFTR) channel conductance that still allows for residual function. Potent...
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MDPI AG
2023-01-01
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| Series: | Journal of Personalized Medicine |
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| Online Access: | https://www.mdpi.com/2075-4426/13/1/102 |
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| author | Mafalda Bacalhau Filipa C. Ferreira Iris A. L. Silva Camilla D. Buarque Margarida D. Amaral Miquéias Lopes-Pacheco |
| author_facet | Mafalda Bacalhau Filipa C. Ferreira Iris A. L. Silva Camilla D. Buarque Margarida D. Amaral Miquéias Lopes-Pacheco |
| author_sort | Mafalda Bacalhau |
| collection | DOAJ |
| description | The R334W (c.1000C>T, p.Arg334Trp) is a rare cystic fibrosis (CF)-causing mutation for which no causal therapy is currently approved. This mutation leads to a significant reduction of CF transmembrane conductance regulator (CFTR) channel conductance that still allows for residual function. Potentiators are small molecules that interact with CFTR protein at the plasma membrane to enhance CFTR-dependent chloride secretion, representing thus pharmacotherapies targeting the root cause of the disease. Here, we generated a new CF bronchial epithelial (CFBE) cell line to screen a collection of compounds and identify novel potentiators for R334W-CFTR. The active compounds were then validated by electrophysiological assays and their additive effects in combination with VX-770, genistein, or VX-445 were exploited in this cell line and further confirmed in intestinal organoids. Four compounds (LSO-24, LSO-25, LSO-38, and LSO-77) were active in the functional primary screen and their ability to enhance R334W-CFTR-dependent chloride secretion was confirmed using electrophysiological measurements. <i>In silico</i> ADME analyses demonstrated that these compounds follow Lipinski’s rule of five and are thus suggested to be orally bioavailable. Dose–response relationships revealed nevertheless suboptimal efficacy and weak potency exerted by these compounds. VX-770 and genistein also displayed a small potentiation of R334W-CFTR function, while VX-445 demonstrated no potentiator activity for this mutation. In the R334W-expressing cell line, CFTR function was further enhanced by the combination of LSO-24, LSO-25, LSO-38, or LSO-77 with VX-770, but not with genistein. The efficacy of potentiator VX-770 combined with active LSO compounds was further confirmed in intestinal organoids (R334W/R334W genotype). Taken together, these molecules were demonstrated to potentiate R334W-CFTR function by a different mechanism than that of VX-770. They may provide a feasible starting point for the design of analogs with improved CFTR-potentiator activity. |
| first_indexed | 2024-03-09T12:03:00Z |
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| id | doaj.art-b778e67c58964fc984daaa27ad692895 |
| institution | Directory Open Access Journal |
| issn | 2075-4426 |
| language | English |
| last_indexed | 2024-03-09T12:03:00Z |
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| series | Journal of Personalized Medicine |
| spelling | doaj.art-b778e67c58964fc984daaa27ad6928952023-11-30T23:02:09ZengMDPI AGJournal of Personalized Medicine2075-44262023-01-0113110210.3390/jpm13010102Additive Potentiation of R334W-CFTR Function by Novel Small MoleculesMafalda Bacalhau0Filipa C. Ferreira1Iris A. L. Silva2Camilla D. Buarque3Margarida D. Amaral4Miquéias Lopes-Pacheco5Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon, PortugalBiosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon, PortugalBiosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon, PortugalDepartment of Chemistry, Pontifical Catholic University of Rio de Janeiro, Rio de Janeiro 22541-041, BrazilBiosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon, PortugalBiosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon, PortugalThe R334W (c.1000C>T, p.Arg334Trp) is a rare cystic fibrosis (CF)-causing mutation for which no causal therapy is currently approved. This mutation leads to a significant reduction of CF transmembrane conductance regulator (CFTR) channel conductance that still allows for residual function. Potentiators are small molecules that interact with CFTR protein at the plasma membrane to enhance CFTR-dependent chloride secretion, representing thus pharmacotherapies targeting the root cause of the disease. Here, we generated a new CF bronchial epithelial (CFBE) cell line to screen a collection of compounds and identify novel potentiators for R334W-CFTR. The active compounds were then validated by electrophysiological assays and their additive effects in combination with VX-770, genistein, or VX-445 were exploited in this cell line and further confirmed in intestinal organoids. Four compounds (LSO-24, LSO-25, LSO-38, and LSO-77) were active in the functional primary screen and their ability to enhance R334W-CFTR-dependent chloride secretion was confirmed using electrophysiological measurements. <i>In silico</i> ADME analyses demonstrated that these compounds follow Lipinski’s rule of five and are thus suggested to be orally bioavailable. Dose–response relationships revealed nevertheless suboptimal efficacy and weak potency exerted by these compounds. VX-770 and genistein also displayed a small potentiation of R334W-CFTR function, while VX-445 demonstrated no potentiator activity for this mutation. In the R334W-expressing cell line, CFTR function was further enhanced by the combination of LSO-24, LSO-25, LSO-38, or LSO-77 with VX-770, but not with genistein. The efficacy of potentiator VX-770 combined with active LSO compounds was further confirmed in intestinal organoids (R334W/R334W genotype). Taken together, these molecules were demonstrated to potentiate R334W-CFTR function by a different mechanism than that of VX-770. They may provide a feasible starting point for the design of analogs with improved CFTR-potentiator activity.https://www.mdpi.com/2075-4426/13/1/102CFTR modulatorco-potentiationcystic fibrosisdrug discoveryintestinal organoidspotentiator |
| spellingShingle | Mafalda Bacalhau Filipa C. Ferreira Iris A. L. Silva Camilla D. Buarque Margarida D. Amaral Miquéias Lopes-Pacheco Additive Potentiation of R334W-CFTR Function by Novel Small Molecules Journal of Personalized Medicine CFTR modulator co-potentiation cystic fibrosis drug discovery intestinal organoids potentiator |
| title | Additive Potentiation of R334W-CFTR Function by Novel Small Molecules |
| title_full | Additive Potentiation of R334W-CFTR Function by Novel Small Molecules |
| title_fullStr | Additive Potentiation of R334W-CFTR Function by Novel Small Molecules |
| title_full_unstemmed | Additive Potentiation of R334W-CFTR Function by Novel Small Molecules |
| title_short | Additive Potentiation of R334W-CFTR Function by Novel Small Molecules |
| title_sort | additive potentiation of r334w cftr function by novel small molecules |
| topic | CFTR modulator co-potentiation cystic fibrosis drug discovery intestinal organoids potentiator |
| url | https://www.mdpi.com/2075-4426/13/1/102 |
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