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In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study

BACKGROUND: Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, which acts as a chloride channel activated by cyclic AMP (cAMP). The most frequent mutation found in 70% of CF patients is F508del, while premature sto...

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Autores principales: Sermet-Gaudelus, Isabelle, Renouil, Michel, Fajac, Anne, Bidou, Laure, Parbaille, Bastien, Pierrot, Sébastien, Davy, Nolwen, Bismuth, Elise, Reinert, Philippe, Lenoir, Gérard, Lesure, Jean François, Rousset, Jean Pierre, Edelman, Aleksander
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1852113/
https://www.ncbi.nlm.nih.gov/pubmed/17394637
http://dx.doi.org/10.1186/1741-7015-5-5
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author Sermet-Gaudelus, Isabelle
Renouil, Michel
Fajac, Anne
Bidou, Laure
Parbaille, Bastien
Pierrot, Sébastien
Davy, Nolwen
Bismuth, Elise
Reinert, Philippe
Lenoir, Gérard
Lesure, Jean François
Rousset, Jean Pierre
Edelman, Aleksander
author_facet Sermet-Gaudelus, Isabelle
Renouil, Michel
Fajac, Anne
Bidou, Laure
Parbaille, Bastien
Pierrot, Sébastien
Davy, Nolwen
Bismuth, Elise
Reinert, Philippe
Lenoir, Gérard
Lesure, Jean François
Rousset, Jean Pierre
Edelman, Aleksander
author_sort Sermet-Gaudelus, Isabelle
collection PubMed
description BACKGROUND: Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, which acts as a chloride channel activated by cyclic AMP (cAMP). The most frequent mutation found in 70% of CF patients is F508del, while premature stop mutations are found in about 10% of patients. In vitro aminoglycoside antibiotics (e.g. gentamicin) suppress nonsense mutations located in CFTR permitting translation to continue to the natural termination codon. Pharmacologic suppression of stop mutations within the CFTR may be of benefit to a significant number of patients. Our pilot study was conducted to determine whether intravenous gentamicin suppresses stop codons in CF patients and whether it has clinical benefits. METHODS: A dual gene reporter system was used to determine the gentamicin-induced readthrough level of the most frequent stop mutations within the CFTR in the French population. We investigated readthrough efficiency in response to 10 mg/kg once-daily intravenous gentamicin perfusions in patients with and without stop mutations. Respiratory function, sweat chloride concentration, nasal potential difference (NPD) and CFTR expression in nasal epithelial cells were measured at baseline and after 15 days of treatment. RESULTS: After in vitro gentamicin incubation, the readthrough efficiency for the Y122X mutation was at least five times higher than that for G542X, R1162X, and W1282X. In six of the nine patients with the Y122X mutation, CFTR immunodetection showed protein at the membrane of the nasal epithelial cells and the CFTR-dependent Cl(- )secretion in NPD measurements increased significantly. Respiratory status also improved in these patients, irrespective of the gentamicin sensitivity of the bacteria present in the sputum. Mean sweat chloride concentration decreased significantly and normalised in two patients. Clinical status, NPD and sweat Cl(- )values did not change in the Y122X patients with no protein expression, in patients with the other stop mutations investigated in vitro and those without stop mutations. CONCLUSION: Suppression of stop mutations in the CFTR gene with parenteral gentamicin can be predicted in vitro and is associated with clinical benefit and significant modification of the CFTR-mediated Cl(- )transport in nasal and sweat gland epithelium.
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spelling pubmed-18521132007-04-14 In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study Sermet-Gaudelus, Isabelle Renouil, Michel Fajac, Anne Bidou, Laure Parbaille, Bastien Pierrot, Sébastien Davy, Nolwen Bismuth, Elise Reinert, Philippe Lenoir, Gérard Lesure, Jean François Rousset, Jean Pierre Edelman, Aleksander BMC Med Research Article BACKGROUND: Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, which acts as a chloride channel activated by cyclic AMP (cAMP). The most frequent mutation found in 70% of CF patients is F508del, while premature stop mutations are found in about 10% of patients. In vitro aminoglycoside antibiotics (e.g. gentamicin) suppress nonsense mutations located in CFTR permitting translation to continue to the natural termination codon. Pharmacologic suppression of stop mutations within the CFTR may be of benefit to a significant number of patients. Our pilot study was conducted to determine whether intravenous gentamicin suppresses stop codons in CF patients and whether it has clinical benefits. METHODS: A dual gene reporter system was used to determine the gentamicin-induced readthrough level of the most frequent stop mutations within the CFTR in the French population. We investigated readthrough efficiency in response to 10 mg/kg once-daily intravenous gentamicin perfusions in patients with and without stop mutations. Respiratory function, sweat chloride concentration, nasal potential difference (NPD) and CFTR expression in nasal epithelial cells were measured at baseline and after 15 days of treatment. RESULTS: After in vitro gentamicin incubation, the readthrough efficiency for the Y122X mutation was at least five times higher than that for G542X, R1162X, and W1282X. In six of the nine patients with the Y122X mutation, CFTR immunodetection showed protein at the membrane of the nasal epithelial cells and the CFTR-dependent Cl(- )secretion in NPD measurements increased significantly. Respiratory status also improved in these patients, irrespective of the gentamicin sensitivity of the bacteria present in the sputum. Mean sweat chloride concentration decreased significantly and normalised in two patients. Clinical status, NPD and sweat Cl(- )values did not change in the Y122X patients with no protein expression, in patients with the other stop mutations investigated in vitro and those without stop mutations. CONCLUSION: Suppression of stop mutations in the CFTR gene with parenteral gentamicin can be predicted in vitro and is associated with clinical benefit and significant modification of the CFTR-mediated Cl(- )transport in nasal and sweat gland epithelium. BioMed Central 2007-03-29 /pmc/articles/PMC1852113/ /pubmed/17394637 http://dx.doi.org/10.1186/1741-7015-5-5 Text en Copyright © 2007 Sermet-Gaudelus et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Sermet-Gaudelus, Isabelle
Renouil, Michel
Fajac, Anne
Bidou, Laure
Parbaille, Bastien
Pierrot, Sébastien
Davy, Nolwen
Bismuth, Elise
Reinert, Philippe
Lenoir, Gérard
Lesure, Jean François
Rousset, Jean Pierre
Edelman, Aleksander
In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title_full In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title_fullStr In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title_full_unstemmed In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title_short In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
title_sort in vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1852113/
https://www.ncbi.nlm.nih.gov/pubmed/17394637
http://dx.doi.org/10.1186/1741-7015-5-5
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