medgenМедицинская генетикаMedical Genetics2073-7998Publishing House «Genius Media» LLC10.25557/2073-7998.2024.01.60-68medgen-2411Research ArticleКЛИНИЧЕСКИЙ СЛУЧАЙCLINICAL CASEИзучение редкого варианта гена CFTR c.1329_1350del в гомозиготном состоянии у ребенка с муковисцидозом с помощью функциональных тестовStudy of a rare variant of the CFTR c.1329_1350del gene in a homozygous state in a child with cystic fibrosis using functional testsКондратьеваЕ. И.KondratyevaE. I.

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КрасноваМ. Г.KrasnovaM. G.

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МельяновскаяЮ. Л.MelyanovskayaYu. L.

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ШерманВ. Д.ShermanV. D.

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МокроусоваД. О.MokrousovaD. O.

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ЕфремоваА. С.EfremovaA. S.

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БулатенкоН. В.BulatenkoN. V.

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БухароваТ. Б.BukharovaT. B.

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ГольдштейнД. В.GoldshteinD. V.

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ФГБНУ Медико-генетический научный центр имени академика Н.П. БочковаРоссияResearch Centre for Medical GeneticsRussian Federation2024050320242316068Copyright © Кондратьева Е.И., Краснова М.Г., Мельяновская Ю.Л., Шерман В.Д., Мокроусова Д.О., Ефремова А.С., Булатенко Н.В., Бухарова Т.Б., Гольдштейн Д.В., 20242024Кондратьева Е.И., Краснова М.Г., Мельяновская Ю.Л., Шерман В.Д., Мокроусова Д.О., Ефремова А.С., Булатенко Н.В., Бухарова Т.Б., Гольдштейн Д.В.Kondratyeva E.I., Krasnova M.G., Melyanovskaya Y.L., Sherman V.D., Mokrousova D.O., Efremova A.S., Bulatenko N.V., Bukharova T.B., Goldshtein D.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.medgen-journal.ru/jour/article/view/2411

   Изучение редких вариантов в гене CFTR является актуальным в эпоху таргетной терапии CFTR-модуляторами. В данном исследовании изучена клиническая картина муковисцидоза, а также функция CFTR-канала у гомозиготного носителя редкого патогенного варианта c.1329_1350del (p.Asp443GlufsX19), относящегося к I классу нарушений в гене CFTR. Получена полная корреляция результатов функциональных тестов с фенотипической картиной муковисцидоза и потовым тестом. Метод определения разности кишечных потенциалов продемонстрировал отсутствие функциональной активности хлорного CFTR канала в ректальном биоптате; при стимулировании форсколином органоиды пациента не ответили набуханием, что свидетельствует о полном нарушении образования функционального белка CFTR. Воздействие на кишечные органоиды всеми зарегистрированными для терапии таргетными препаратами не привело к их набуханию, в отличие от контрольной культуры, гомозиготной по F508del, таким образом, хлорный канал пациента с генотипом p.Asp443GlufsX19/p.Asp443GlufsX19 оказался нечувствителен ни к одному CFTR-модулятору.

   The study of rare variants in the CFTR gene is relevant in the era of targeted therapy with CFTR modulators. In this study, we studied the clinical picture of cystic fibrosis, as well as the function of the CFTR channel in a homozygous carrier of a rare pathogenic variant c.1329_1350del (p.Asp443GlufsX19), belonging to class I disorders in the CFTR gene. A complete correlation of the results of functional tests with the phenotypic pattern of CF and data of a sweat test was obtained. The method for determining the difference in intestinal potentials demonstrated the absence of functional activity of the chloride CFTR channel in the rectal biopsy, and when stimulated with forskolin, the patient’s organoids did not swell, which indicates a complete violation of the functional CFTR protein production. Exposure to intestinal organoids by all registered for therapy targeted drugs did not lead to their swelling, unlike the control culture, homozygous for F508del. The chloride channel of a patient with genotype p.Asp443GlufsX19/p.Asp443GlufsX19 turned out to be insensitive to any CFTR modulator.

муковисцидозредкие варианты гена CFTRкишечные органоидыCFTR-модуляторыcystic fibrosisrare variants of the CFTR geneintestinal organoidstargeted therapyCFTR-modulatorsРабота выполнена в рамках государственного задания ФГБНУ «Медико-генетический научный центр имени академика Н. П. Бочкова»The work was carried out under the state assignment for the Research Centre for Medical GeneticsReferencesEgan M.E. Genetics of Cystic Fibrosis. Clin. Chest Med. 2016; 37; 9–16Egan M.E. Genetics of Cystic Fibrosis. Clin. Chest Med. 2016; 37; 9–16Marson F.A.L., Bertuzzo C.S., Ribeiro J.D. Classification of CFTR mutation classes. Lancet Respir. Med. 2016; 4; e37–8Marson F.A.L., Bertuzzo C.S., Ribeiro J.D. Classification of CFTR mutation classes. Lancet Respir. Med. 2016; 4; e37–8Pereira S.V.-N., Ribeiro J.D. Ribeiro A.F., et al. Novel, rare and common pathogenic variants in the CFTR gene screened by high-throughput sequencing technology and predicted by in silico tools. Sci. Rep. 2019; 9; 6234Pereira S.V.-N., Ribeiro J.D. Ribeiro A.F., et al. Novel, rare and common pathogenic variants in the CFTR gene screened by high-throughput sequencing technology and predicted by in silico tools. Sci. Rep. 2019; 9; 6234Noordhoek J., Gulmans V., Van Der Ent K., et al. Intestinal organoids and personalized medicine in cystic fibrosis: A successful patient-oriented research collaboration. Curr. Opin. Pulm. Med. 2016; 22; 610–6Noordhoek J., Gulmans V., Van Der Ent K., et al. Intestinal organoids and personalized medicine in cystic fibrosis: A successful patient-oriented research collaboration. Curr. Opin. Pulm. Med. 2016; 22; 610–6Boj S.F., Vonk A.M., Statia M., et al. Forskolin-induced swelling in intestinal organoids: An in vitro assay for assessing drug response in cystic fibrosis patients. J. Vis. Exp. 2017; 2017; 1–12Boj S.F., Vonk A.M., Statia M., et al. Forskolin-induced swelling in intestinal organoids: An in vitro assay for assessing drug response in cystic fibrosis patients. J. Vis. Exp. 2017; 2017; 1–12Muilwijk D., de Poel E., van Mourik P., et al. Forskolin-induced organoid swelling is associated with long-term cystic fibrosis disease progression. Eur. Respir. J. 2022; 60Muilwijk D., de Poel E., van Mourik P., et al. Forskolin-induced organoid swelling is associated with long-term cystic fibrosis disease progression. Eur. Respir. J. 2022; 60Регистр пациентов с муковисцидозом в Российской Федерации. 2020 год. Под редакцией Е.И. Кондратьевой, С.А. Красовского, М.А. Стариновой, А.Ю. Воронковой, Е.Л. Амелиной, Н.Ю. Каширской, С.Н. Авдеева, С.И. Куцева.: ИД «Медпрактика-М». 2022: 68 с.Registr patsiyentov s mukovistsidozom v Rossiyskoy Federatsii. 2020 god. Pod redaktsiyey Ye.I. Kondrat’yevoy, S.A. Krasovskogo, M.A. Starinovoy, A.Yu. Voronkovoy, Ye.L. Amelinoy, N.Yu. Kashirskoy, S.N. Avdeyeva, S.I. Kutseva [Register of patients with cystic fibrosis in the Russian Federation. 2020 Edited by E.I. Kondratyeva, S.A. Krasovsky, M.A. Starinova, A.Yu. Voronkova, E.L. Amelina, N.Yu. Kashirskaya, S.N. Avdeev, S.I. Kutsev]. ID «Medpraktika-M» [Publishing House “Medpraktika-M”]. 2022 ; 68 p. (In Russ.)Castellani C., Duff A.J.A., Bell S.C., et al. ECFS best practice guidelines: the 2018 revision. J. Cyst. Fibros. 2018; 17; 153–78Castellani C., Duff A.J.A., Bell S.C., et al. ECFS best practice guidelines: the 2018 revision. J. Cyst. Fibros. 2018; 17; 153–78Клинические рекомендации. Кистозный Фиброз (Муковисцидоз). 2021 https://cr.minzdrav.gov.ru/schema/372_2Klinicheskiye rekomendatsii. Kistoznyy Fibroz (Mukovistsidoz). 2021 [Clinical recommendations. Cystic Fibrosis (Cystic Fibrosis). 2021] https://cr.minzdrav.gov.ru/schema/372_2 (In Russ.)Dekkers J.F., van der Ent C.K., Beekman J.M. Novel opportunities for CFTR-targeting drug development using organoids. Rare Dis. 2013; 1; 1–6Dekkers J.F., van der Ent C.K., Beekman J.M. Novel opportunities for CFTR-targeting drug development using organoids. Rare Dis. 2013; 1; 1–6Vonk A.M., van Mourik P., Ramalho A.S., et al. Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids. STAR Protoc. 2020; 1; 1–31Vonk A.M., van Mourik P., Ramalho A.S., et al. Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids. STAR Protoc. 2020; 1; 1–31Dekkers J.F., Berkers G., Kruisselbrink E., et al. Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis. Sci. Transl. Med. 2016; 8Dekkers J.F., Berkers G., Kruisselbrink E., et al. Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis. Sci. Transl. Med. 2016; 8Ramalho A.S., Förstová E., Vonk A.M., et al. Correction of CFTR function in intestinal organoids to guide treatment of cystic fibrosis. Eur. Respir. J. 2021; 57Ramalho A.S., Förstová E., Vonk A.M., et al. Correction of CFTR function in intestinal organoids to guide treatment of cystic fibrosis. Eur. Respir. J. 2021; 57Melyanovskaya J.L., Kondratieva E.I., Kutsev S.I. Determination of reference values for the method of intestinal current measurement in the Russian Federation. Med. News North Caucasus 2020; 15Melyanovskaya J.L., Kondratieva E.I., Kutsev S.I. Determination of reference values for the method of intestinal current measurement in the Russian Federation. Med. News North Caucasus 2020; 15Guimbellot J., Sharma J., Rowe S.M. Toward inclusive therapy with CFTR modulators: Progress and challenges. Pediatr. Pulmonol. 2017; 52; S4–14Guimbellot J., Sharma J., Rowe S.M. Toward inclusive therapy with CFTR modulators: Progress and challenges. Pediatr. Pulmonol. 2017; 52; S4–14Derichs N., Sanz J., Von Kanel T., et al. Intestinal current measurement for diagnostic classification of patients with questionable cystic fibrosis: Validation and reference data. Thorax 2010; 65; 594–9Derichs N., Sanz J., Von Kanel T., et al. Intestinal current measurement for diagnostic classification of patients with questionable cystic fibrosis: Validation and reference data. Thorax 2010; 65; 594–9Clancy J.P., Szczesniak R.D., Ashlock M.A., et al. Multicenter Intestinal Current Measurements in Rectal Biopsies from CF and Non-CF Subjects to Monitor CFTR Function. PLoS One 2013; 8; 1–13Clancy J.P., Szczesniak R.D., Ashlock M.A., et al. Multicenter Intestinal Current Measurements in Rectal Biopsies from CF and Non-CF Subjects to Monitor CFTR Function. PLoS One 2013; 8; 1–13Kondratyeva E., Melyanovskaya Y., Efremova A., et al. Clinical and Genetic Characteristics of a Patient with Cystic Fibrosis with a Complex Allele [E217G;G509D] and Functional Evaluation of the CFTR Channel. Genes (Basel). 2023; 14; 1705Kondratyeva E., Melyanovskaya Y., Efremova A., et al. Clinical and Genetic Characteristics of a Patient with Cystic Fibrosis with a Complex Allele [E217G;G509D] and Functional Evaluation of the CFTR Channel. Genes (Basel). 2023; 14; 1705Kondratyeva E., Melyanovskaya Y., Bulatenko N., et al. Clinical and Functional Characteristics of the E92K CFTR Gene Variant in the Russian and Turkish Population of People with Cystic Fibrosis. Int. J. Mol. Sci. 2023; 24; 6351Kondratyeva E., Melyanovskaya Y., Bulatenko N., et al. Clinical and Functional Characteristics of the E92K CFTR Gene Variant in the Russian and Turkish Population of People with Cystic Fibrosis. Int. J. Mol. Sci. 2023; 24; 6351Kondratyeva E., Bulatenko N. Melyanovskaya Y., et al. Personalized Selection of a CFTR Modulator for a Patient with a Complex Allele [L467F;F508del]. Curr. Issues Mol. Biol. 2022; 44; 5126–38Kondratyeva E., Bulatenko N. Melyanovskaya Y., et al. Personalized Selection of a CFTR Modulator for a Patient with a Complex Allele [L467F;F508del]. Curr. Issues Mol. Biol. 2022; 44; 5126–38Kondratyeva E., Bukharova T., Efremova A., et al. 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. Genes (Basel). 2021; 12.Kondratyeva E., Bukharova T., Efremova A., et al. 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. Genes (Basel). 2021; 12

The authors declare that there are no conflicts of interest present.