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Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant
Cystic fibrosis (CF) is the most common monogenic autosomal recessive disease, associated with pathogenic variants in the CFTR gene. The splicing variant c.3140-16T>A (3272-16T>A) has been described previously and, according to the Russian CF Patients Registry, occurs with a frequency of 0.34%...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229552/ https://www.ncbi.nlm.nih.gov/pubmed/34071719 http://dx.doi.org/10.3390/genes12060837 |
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author | Kondratyeva, Elena Bukharova, Tatyana Efremova, Anna Melyanovskaya, Yuliya Bulatenko, Natalia Davydenko, Ksenia Filatova, Alexandra Skoblov, Mikhail Krasovsky, Stanislav Petrova, Nika Polyakov, Alexander Adyan, Tagui Amelina, Elena Shadrina, Vera Zhekaite, Elena Zodbinova, Aysa Chernyak, Alexander Zinchenko, Rena Kutsev, Sergei Goldshtein, Dmitry |
author_facet | Kondratyeva, Elena Bukharova, Tatyana Efremova, Anna Melyanovskaya, Yuliya Bulatenko, Natalia Davydenko, Ksenia Filatova, Alexandra Skoblov, Mikhail Krasovsky, Stanislav Petrova, Nika Polyakov, Alexander Adyan, Tagui Amelina, Elena Shadrina, Vera Zhekaite, Elena Zodbinova, Aysa Chernyak, Alexander Zinchenko, Rena Kutsev, Sergei Goldshtein, Dmitry |
author_sort | Kondratyeva, Elena |
collection | PubMed |
description | Cystic fibrosis (CF) is the most common monogenic autosomal recessive disease, associated with pathogenic variants in the CFTR gene. The splicing variant c.3140-16T>A (3272-16T>A) has been described previously and, according to the Russian CF Patients Registry, occurs with a frequency of 0.34%. The phenotypic features of CF patients with the c.3140-16T>A variant were compared with those of patients with the genotype F508del/F508del. Patients with the allele c.3140-16T>A had higher average age and age at diagnosis, and the allele was present in a greater proportion of adults. Patients carrying the c.3140-16T>A allele were characterised by better physical development indicators, both in adults and in children, had preserved pancreatic function, as well as the absence of a number of complications, and required pancreatic enzyme replacement therapy less often than patients with the F508del/F508del genotype. Sweat test values also were lower in patients with the c.3140-16T>A genotype. According to the results of clinical and laboratory studies, the phenotype of patients with the genetic variant c.3140-16T>A can be considered “mild”. Functional CFTR protein activity in the presence of c.3140-16T>A was evaluated using intestinal current measurements (ICM) and the forskolin-induced swelling assay on organoids obtained from patients’ rectal biopsies. c.3140-16T>A had high residual CFTR channel activity and was amenable to effective pharmacological correction with thea VX-770 potentiator. To evaluate the effect of the variant on CFTR pre-mRNA splicing we performed a minigene assay, as well as RT-PCR analysis of RNA isolated from the nasal epithelium and rectal biopsy of patients. We showed that the c.3140-16T>A variant creates a novel acceptor AG dinucleotide within CFTR intron 19, resulting in a 14-nucleotide extension of exon 20. This frameshift produces a premature termination codon and triggers mRNA degradation by the nonsense-mediated decay (NMD) mechanism. Moreover, we observed that the c.3140-16T>A allele could produce a residual amount of normally spliced transcript, thus explaining the patient’s mild phenotype. |
format | Online Article Text |
id | pubmed-8229552 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-82295522021-06-26 Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant Kondratyeva, Elena Bukharova, Tatyana Efremova, Anna Melyanovskaya, Yuliya Bulatenko, Natalia Davydenko, Ksenia Filatova, Alexandra Skoblov, Mikhail Krasovsky, Stanislav Petrova, Nika Polyakov, Alexander Adyan, Tagui Amelina, Elena Shadrina, Vera Zhekaite, Elena Zodbinova, Aysa Chernyak, Alexander Zinchenko, Rena Kutsev, Sergei Goldshtein, Dmitry Genes (Basel) Article Cystic fibrosis (CF) is the most common monogenic autosomal recessive disease, associated with pathogenic variants in the CFTR gene. The splicing variant c.3140-16T>A (3272-16T>A) has been described previously and, according to the Russian CF Patients Registry, occurs with a frequency of 0.34%. The phenotypic features of CF patients with the c.3140-16T>A variant were compared with those of patients with the genotype F508del/F508del. Patients with the allele c.3140-16T>A had higher average age and age at diagnosis, and the allele was present in a greater proportion of adults. Patients carrying the c.3140-16T>A allele were characterised by better physical development indicators, both in adults and in children, had preserved pancreatic function, as well as the absence of a number of complications, and required pancreatic enzyme replacement therapy less often than patients with the F508del/F508del genotype. Sweat test values also were lower in patients with the c.3140-16T>A genotype. According to the results of clinical and laboratory studies, the phenotype of patients with the genetic variant c.3140-16T>A can be considered “mild”. Functional CFTR protein activity in the presence of c.3140-16T>A was evaluated using intestinal current measurements (ICM) and the forskolin-induced swelling assay on organoids obtained from patients’ rectal biopsies. c.3140-16T>A had high residual CFTR channel activity and was amenable to effective pharmacological correction with thea VX-770 potentiator. To evaluate the effect of the variant on CFTR pre-mRNA splicing we performed a minigene assay, as well as RT-PCR analysis of RNA isolated from the nasal epithelium and rectal biopsy of patients. We showed that the c.3140-16T>A variant creates a novel acceptor AG dinucleotide within CFTR intron 19, resulting in a 14-nucleotide extension of exon 20. This frameshift produces a premature termination codon and triggers mRNA degradation by the nonsense-mediated decay (NMD) mechanism. Moreover, we observed that the c.3140-16T>A allele could produce a residual amount of normally spliced transcript, thus explaining the patient’s mild phenotype. MDPI 2021-05-28 /pmc/articles/PMC8229552/ /pubmed/34071719 http://dx.doi.org/10.3390/genes12060837 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kondratyeva, Elena Bukharova, Tatyana Efremova, Anna Melyanovskaya, Yuliya Bulatenko, Natalia Davydenko, Ksenia Filatova, Alexandra Skoblov, Mikhail Krasovsky, Stanislav Petrova, Nika Polyakov, Alexander Adyan, Tagui Amelina, Elena Shadrina, Vera Zhekaite, Elena Zodbinova, Aysa Chernyak, Alexander Zinchenko, Rena Kutsev, Sergei Goldshtein, Dmitry Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title | Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title_full | Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title_fullStr | Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title_full_unstemmed | Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title_short | Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T>A and Functional Analysis of this Variant |
title_sort | health characteristics of patients with cystic fibrosis whose genotype includes a variant of the nucleotide sequence c.3140-16t>a and functional analysis of this variant |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229552/ https://www.ncbi.nlm.nih.gov/pubmed/34071719 http://dx.doi.org/10.3390/genes12060837 |
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