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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%...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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.
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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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