Cystic fibrosis: modern possibilities of diagnosis and treatment based on molecular pathogenesis

Background. Cystic fibrosis is an inherited autosomal recessive disease. Modern methods of DNA diagnosis open up the possibility to develop new effective methods of cystic fibrosis treatment, which are based on the impact on the molecular pathogenesis of the disease. A personalized approach to medical care for patients with cystic fibrosis, taking into account the genetic profile with the use of new DNA diagnostic technologies has raised the provision of medical and genetic counseling for burdened families to a new level and more effective treatment measures. Aim. Implementation of a program of modern molecular genetic diagnosis of inherited diseases using targeted therapy for cystic fibrosis. Methods. The data for the entire period of neonatal screening for cystic fibrosis in the Republic of Bashkortostan were analyzed. In 2021 there were 94 patients on the cystic fibrosis registry in the Republic of Bashkortostan. The majority of patients required DNA diagnosis to verify mutations in the CFTR gene with subsequent prescription of tragent therapy. In the first phase of the study, 94 patients with cystic fibrosis as well as their relatives were analyzed using multiplex ligase-dependent probe amplification (SALSA MLPA Probemix P091 CFTR, MRC Holland). In the next step, all exons of the CFTR gene were sequenced using next-generation sequencing (NGS) and Sanger methods. Results. A search for mutations was performed in 94 probands and 36 variants of changes in the CFTR gene with pathogenic significance for the development of cystic fibrosis were identified. No mutations were found on 2.6% of chromosomes, and mutations were identified on 97.3% of chromosomes. In 89 patients both mutations were identified (83.66%), in 5 probands only one heterozygous mutation was found. One patient was diagnosed as having сystic fibrosis. Conclusion. The development and implementation of the latest sequencing technologies have significantly expanded knowledge about the molecular mechanisms of cystic fibrosis, which contributes to the development of new approaches to therapy and new research.

cystic fibrosis, newborn screening, DNA diagnostics, CFTR, targeted therapy

1. Регистр больных муковисцидозом в Российской Федерации. 2018 год. Под редакцией Е.Л. Амелиной, Н.Ю. Каширской, Е.И. Кондратьевой, С.А. Красовского, М.А. Стариновой, А.Ю. Воронковой.- М.: ИД «МЕДПРАКТИКА-М», 2020, 68 с.

2. Butnariu L.I., Țarcă E., Cojocaru E., Rusu C., Moisă Ș.M., Leon Constantin M.M., Gorduza E.V., Trandafir L.M. Genetic Modifying Factors of Cystic Fibrosis Phenotype: A Challenge for Modern Medicine J. Clin. Med. 2021, 10(24), 5821; https://doi.org/10.3390/jcm10245821

3. Муковисцидоз. Издание 2-е., переработанное и дополненное (под редакцией Н.Ю.Каширской, Н.И.Капранова и Е.И.Кондратьевой). - М.: ИД «МЕДПРАКТИКА-М», 2021, 680 с.

4. Капранов Н.И., Каширская Н.Ю., Ашерова И.К., Кондратьева Е.И., Шерман В.Д. Исторические и современные аспекты муковисцидоза в России. Педиатрическая фармакология. 2013;10(6):53-60. https://doi.org/10.15690/pf.v10i6.896

5. Национальный консенсус (2 -е издание) «Муковисцидоз: определение, диагностические критерии, терапия»2018/ Под редакцией Е.И. Кондратьевой, Н.Ю. Каширской, Н.И. Капранова - М.: ООО «Компания БОРГЕС»., 2018, 356 с.

6. Капранов Н.И., Каширская Н.Ю. Муковисцидоз. Современные достижения и актуальные проблемы. Методические рекомендации. Издание 3-е, переработанное и дополненное. М.: ООО «4ТЕ Арт»; 2008. 124 с.

7. Толстова В.Д., Каширская Н.Ю., Капранов Н.И. Массовый скрининг новорожденных на муковисцидоз в России. Фарматека. 2008;1:38-43

8. Wainwright C.E., Elborn J.S., Ramsey B.W., Marigowda G., Huang X., Cipolli M., Colombo C., Davies J.C., De Boeck K., Flume P.A., Konstan M.W., McColley S.A., McCoy K., McKone E.F., Munck A., Ratjen F., Rowe S.M., Waltz D., Boyle M.P.; TRAFFIC Study Group; TRANSPORT Study Group. Lumacaftor-Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del CFTR. N Engl J Med. 2015;373(3):220-31. doi: 10.1056/NEJMoa1409547.

9. Shaw M., Khan U., Clancy J.P., Donaldson S.H., Sagel S.D., Rowe S.M., Ratjen F. Changes in LCI in F508del/F508del patients treated with lumacaftor/ivacaftor: Results from the prospect study. J Cyst Fibros. 2020;19(6):931-933. doi: 10.1016/j.jcf.2020.05.010.

10. Chevalier B., Hinzpeter A. The influence of CFTR complex alleles on precision therapy of cystic fibrosis. J Cyst Fibros. 2020;19(Suppl 1):S15-S18. DOI:10.1016/j.jcf.2019.12.008

11. Baatallah N., Bitam S., Martin N., et al. Cis variants identified in F508del complex alleles modulate CFTR channel rescue by small molecules. Hum Mutat. 2018; 39:506-14. DOI: 10.1002/humu.23389

12. Кондратьева Е.И., Амелина Е.Л., Чернуха М.Ю., и др. Обзор клинических рекомендаций «Кистозный фиброз (муковисцидоз)» 2020. Пульмонология. 2021; 31(2):135-146. DOI: 10.18093/0869-0189-2021-31-2-135-146

13. Амелина Е.Л., Ефремова А.С., Мельяновская Ю.Л., и др. Использование функциональных тестов для оценки остаточной активности канала CFTR и индивидуального подбора эффективных CFTR-модуляторов для лечения пациентов с муковисцидозом с «мягким» и «тяжёлым» генетическими вариантами. Пульмонология. 2021;31(2):167-176

14. Ефремова А.С., Мельяновская Ю.Л., Булатенко Н.В., и др. Описание редких аллелей гена CFTR при муковисцидозе с помощью функциональных тестов и форсколинового теста на ректальных органоидах. Пульмонология. 2021;31(2):178-188

15. Куцев С.И., Ижевская В.Л., Кондратьева Е.И. Таргетная терапия при муковисцидозе. Пульмонология. 2021; 31(2): 226-236. DOI:10.18093/0869-0189-2021-31-2-226-236