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Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis

CFTR modulators have revolutionized the treatment of individuals with cystic fibrosis (CF) by improving the function of existing protein. Unfortunately, almost half of the disease-causing variants in CFTR are predicted to introduce premature termination codons (PTC) thereby causing absence of full-l...

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Autores principales: Sharma, Neeraj, Evans, Taylor A., Pellicore, Matthew J., Davis, Emily, Aksit, Melis A., McCague, Allison F., Joynt, Anya T., Lu, Zhongzhu, Han, Sangwoo T., Anzmann, Arianna F., Lam, Anh-Thu N., Thaxton, Abigail, West, Natalie, Merlo, Christian, Gottschalk, Laura B., Raraigh, Karen S., Sosnay, Patrick R., Cotton, Calvin U., Cutting, Garry R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267994/
https://www.ncbi.nlm.nih.gov/pubmed/30444886
http://dx.doi.org/10.1371/journal.pgen.1007723
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author Sharma, Neeraj
Evans, Taylor A.
Pellicore, Matthew J.
Davis, Emily
Aksit, Melis A.
McCague, Allison F.
Joynt, Anya T.
Lu, Zhongzhu
Han, Sangwoo T.
Anzmann, Arianna F.
Lam, Anh-Thu N.
Thaxton, Abigail
West, Natalie
Merlo, Christian
Gottschalk, Laura B.
Raraigh, Karen S.
Sosnay, Patrick R.
Cotton, Calvin U.
Cutting, Garry R.
author_facet Sharma, Neeraj
Evans, Taylor A.
Pellicore, Matthew J.
Davis, Emily
Aksit, Melis A.
McCague, Allison F.
Joynt, Anya T.
Lu, Zhongzhu
Han, Sangwoo T.
Anzmann, Arianna F.
Lam, Anh-Thu N.
Thaxton, Abigail
West, Natalie
Merlo, Christian
Gottschalk, Laura B.
Raraigh, Karen S.
Sosnay, Patrick R.
Cotton, Calvin U.
Cutting, Garry R.
author_sort Sharma, Neeraj
collection PubMed
description CFTR modulators have revolutionized the treatment of individuals with cystic fibrosis (CF) by improving the function of existing protein. Unfortunately, almost half of the disease-causing variants in CFTR are predicted to introduce premature termination codons (PTC) thereby causing absence of full-length CFTR protein. We hypothesized that a subset of nonsense and frameshift variants in CFTR allow expression of truncated protein that might respond to FDA-approved CFTR modulators. To address this concept, we selected 26 PTC-generating variants from four regions of CFTR and determined their consequences on CFTR mRNA, protein and function using intron-containing minigenes expressed in 3 cell lines (HEK293, MDCK and CFBE41o-) and patient-derived conditionally reprogrammed primary nasal epithelial cells. The PTC-generating variants fell into five groups based on RNA and protein effects. Group A (reduced mRNA, immature (core glycosylated) protein, function <1% (n = 5)) and Group B (normal mRNA, immature protein, function <1% (n = 10)) variants were unresponsive to modulator treatment. However, Group C (normal mRNA, mature (fully glycosylated) protein, function >1% (n = 5)), Group D (reduced mRNA, mature protein, function >1% (n = 5)) and Group E (aberrant RNA splicing, mature protein, function > 1% (n = 1)) variants responded to modulators. Increasing mRNA level by inhibition of NMD led to a significant amplification of modulator effect upon a Group D variant while response of a Group A variant was unaltered. Our work shows that PTC-generating variants should not be generalized as genetic ‘nulls’ as some may allow generation of protein that can be targeted to achieve clinical benefit.
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spelling pubmed-62679942018-12-19 Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis Sharma, Neeraj Evans, Taylor A. Pellicore, Matthew J. Davis, Emily Aksit, Melis A. McCague, Allison F. Joynt, Anya T. Lu, Zhongzhu Han, Sangwoo T. Anzmann, Arianna F. Lam, Anh-Thu N. Thaxton, Abigail West, Natalie Merlo, Christian Gottschalk, Laura B. Raraigh, Karen S. Sosnay, Patrick R. Cotton, Calvin U. Cutting, Garry R. PLoS Genet Research Article CFTR modulators have revolutionized the treatment of individuals with cystic fibrosis (CF) by improving the function of existing protein. Unfortunately, almost half of the disease-causing variants in CFTR are predicted to introduce premature termination codons (PTC) thereby causing absence of full-length CFTR protein. We hypothesized that a subset of nonsense and frameshift variants in CFTR allow expression of truncated protein that might respond to FDA-approved CFTR modulators. To address this concept, we selected 26 PTC-generating variants from four regions of CFTR and determined their consequences on CFTR mRNA, protein and function using intron-containing minigenes expressed in 3 cell lines (HEK293, MDCK and CFBE41o-) and patient-derived conditionally reprogrammed primary nasal epithelial cells. The PTC-generating variants fell into five groups based on RNA and protein effects. Group A (reduced mRNA, immature (core glycosylated) protein, function <1% (n = 5)) and Group B (normal mRNA, immature protein, function <1% (n = 10)) variants were unresponsive to modulator treatment. However, Group C (normal mRNA, mature (fully glycosylated) protein, function >1% (n = 5)), Group D (reduced mRNA, mature protein, function >1% (n = 5)) and Group E (aberrant RNA splicing, mature protein, function > 1% (n = 1)) variants responded to modulators. Increasing mRNA level by inhibition of NMD led to a significant amplification of modulator effect upon a Group D variant while response of a Group A variant was unaltered. Our work shows that PTC-generating variants should not be generalized as genetic ‘nulls’ as some may allow generation of protein that can be targeted to achieve clinical benefit. Public Library of Science 2018-11-16 /pmc/articles/PMC6267994/ /pubmed/30444886 http://dx.doi.org/10.1371/journal.pgen.1007723 Text en © 2018 Sharma et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sharma, Neeraj
Evans, Taylor A.
Pellicore, Matthew J.
Davis, Emily
Aksit, Melis A.
McCague, Allison F.
Joynt, Anya T.
Lu, Zhongzhu
Han, Sangwoo T.
Anzmann, Arianna F.
Lam, Anh-Thu N.
Thaxton, Abigail
West, Natalie
Merlo, Christian
Gottschalk, Laura B.
Raraigh, Karen S.
Sosnay, Patrick R.
Cotton, Calvin U.
Cutting, Garry R.
Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title_full Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title_fullStr Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title_full_unstemmed Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title_short Capitalizing on the heterogeneous effects of CFTR nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
title_sort capitalizing on the heterogeneous effects of cftr nonsense and frameshift variants to inform therapeutic strategy for cystic fibrosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267994/
https://www.ncbi.nlm.nih.gov/pubmed/30444886
http://dx.doi.org/10.1371/journal.pgen.1007723
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