ketendoКлиническая и экспериментальная тиреоидологияClinical and experimental thyroidology1995-54722310-3787Endocrinology Research Centre10.14341/ket9556ketendo-9556Research ArticleОригинальные исследованияOriginal StudiesМолекулярно-генетические основы дисгенезии щитовидной железыStudy of molecular basis of thyroid dysgenesishttps://orcid.org/0000-0003-0412-7140МакрецкаяНина АлексеевнаMakretskayaNina A.

научный сотрудник отделения наследственных эндокринопатий

MD, research associate

makretskayan@gmail.comhttps://orcid.org/0000-0001-9621-5732Безлепкина Ольга БорисовнаBezlepkinaOlga B.

д.м.н., профессор, заместитель директора Центра по научно-лечебной работе

MD, PhD, Professor

olgabezlepkina@mail.ruhttps://orcid.org/0000-0001-7736-5372КолодкинаАнна АлександровнаKolodkinaAnna A.

к.м.н., старший научный сотрудник отделения наследственных эндокринопатий

MD, PhD, senior research associate

anna_kolodkina@mail.ruhttps://orcid.org/0000-0002-5578-5242КияевАлексей ВасильевичKiyaevAlexey V.

д.м.н., доцент

MD, PhD, assistant professor

thyroend@mail.ruhttps://orcid.org/0000-0003-1107-362XВасильевЕвгений ВитальевичVasilyevEvgeny V.

к.б.н., старший научный сотрудник отделения наследственных эндокринопатий

PhD, senior research associate

vas-evg@yandex.ruhttps://orcid.org/0000-0002-0520-9132ПетровВасилий МихайловичPetrovVasily M.

к.б.н., старший научный сотрудник отделения наследственных эндокринопатий

PhD, senior research associate

petrov.vasiliy@gmail.comhttps://orcid.org/0000-0002-4743-4661ЧикулаеваОльга АлександровнаChikulaevaOlga A.

к.м.н., заместитель директора Института детской эндокринологии по лечебной работе

MD, PhD

chikulaeva.olga@gmail.comhttps://orcid.org/0000-0003-2599-0867МалиевскийОлег АртуровичMalievskyOleg A.

д.м.н., профессор

MD, PhD, Professor

malievsky@list.ruhttps://orcid.org/0000-0002-8175-7886ДедовИван ИвановичDedovIvan I.

д.м.н., профессор, академик РАН, директор центра

MD, PhD, Professor

dedov@endocrincentr.ruhttps://orcid.org/0000-0001-8500-4841ТюльпаковАнатолий НиколаевичTyulpakovAnatoliy N.

д.м.н., заведующий отделения наследственных эндокринопатий

MD, PhD

ant@endocrincentr.ru<p>ФГБУ "Национальный медицинский исследовательский центр эндокринологии" Минздрава России</p>Россия<p>Endocrinology Research Centre</p>Russian Federation<p>ФГБОУ ВО &laquo;Уральский Государственный Медицинский Университет&raquo; Минздрава России</p>Россия<p>Ural State Medical University</p>Russian Federation<p>ГБУЗ РБ &laquo;Республиканская детская клиническая больница&raquo;</p>Россия<p>Republican Children&rsquo;s Clinical Hospital</p>Russian Federation2018230820181426471Copyright &#x00A9; Макрецкая Н.А., Безлепкина  О.Б., Колодкина А.А., Кияев А.В., Васильев Е.В., Петров В.М., Чикулаева О.А., Малиевский О.А., Дедов И.И., Тюльпаков А.Н., 20182018Макрецкая Н.А., Безлепкина  О.Б., Колодкина А.А., Кияев А.В., Васильев Е.В., Петров В.М., Чикулаева О.А., Малиевский О.А., Дедов И.И., Тюльпаков А.Н.Makretskaya N.A., Bezlepkina O.B., Kolodkina A.A., Kiyaev A.V., Vasilyev E.V., Petrov V.M., Chikulaeva O.A., Malievsky O.A., Dedov I.I., Tyulpakov A.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.cet-endojournals.ru/jour/article/view/9556

Врожденный гипотиреоз – гетерогенная группа заболеваний, объединенных общим признаком – снижением функции щитовидной железы (ЩЖ) к моменту рождения. 80–85% случаев заболевания обусловлены различными вариантами нарушения органогенеза ЩЖ. На сегодняшний день в литературе описано 5 генов: TSHR, PAX8, FOXE1, NKX2-1, NKX2-5, задействованных в патогенезе дисгенезии щитовидной железы.

Цель

Цель. Оценить частоту мутаций в генах TSHR, PAX8, FOXE1, NKX2-1, NKX2-5 среди пациентов с тяжелым врожденным гипотиреозом.

Методы

Методы. В исследование включен 161 пациент с врожденным гипотиреозом (64 мальчика, 97 девочек) с уровнем тиреотропного гормона по данным скрининга или ретестирования более 90 мМЕ/л. При ультразвуковом исследовании ЩЖ у 138 обследуемых диагностированы различные варианты дисгенезии, у 23 пациентов объем железы соответствовал нормальным значениям относительно площади поверхности тела. Для молекулярно-генетического анализа применялся метод высокопроизводительного параллельного секвенирования. Секвенирование осуществлялось на полупроводниковом секвенаторе PGM (Ion Torrent, Life Technologies, США) с использованием панели праймеров “Гипотиреоз” (Custom DNA Panel). Оценка патогенности мутаций осуществлялась согласно последним международным рекомендациям (ACMG, 2015).

Результаты

Результаты. Мутации в генах, приводящие к дисгенезии щитовидной железы, были выявлены у 13 пациентов (8,1%, 13/161): TSHR, n = 6; NKX2-1, n = 3; NKX2-5, n = 1; PAX8, n = 3; FOXE1, n = 0.

Заключение

Заключение. Мутации в генах, обусловливающие развитие дисгенезии щитовидной железы, являются редкой патологией. Среди наших пациентов наибольшее количество мутаций выявлено в гене TSHR.

Congenital hypothyroidism is a heterogeneous group of diseases, which is manifested by loss of function of the thyroid gland that affects infants from birth. 80–85% of cases are due to different types of thyroid dysgenesis. 5 genes have been described that are involved in the pathogenesis of thyroid dysgenesis: TSHR, PAX8, FOXE1, NKX2-1, NKX2-5.

Aims

Aims. To evaluate the prevalence of mutations in the genes TSHR, PAX8, FOXE1, NKX2-1, NKX2-5 among patients with severe congenital hypothyroidism.

Materials and methods

Materials and methods. 161 patients (64 boys, 97 girls) with congenital hypothyroidism (TSH levels at neonatal screening or retesting greater than 90 mU/l) were included in the study. 138 subjects had different variants of thyroid dysgenesis, and 23 patients had normal volume of the gland. A next generation sequencing was used for molecular-genetic analysis. Sequencing was performed using PGM semiconductor sequencer (Ion Torrent, Life Technologies, USA) and a panel “Hypothyroidism” (Custom DNA Panel). Assessment of the pathogenicity of sequence variants were carried out according to the latest international guidelines (ACMG, 2015).

Results

Results. 13 patients had variants in thyroid dysgenesis genes (8,1%, 13/161): TSHR, n = 6; NKX2-1, n = 3; NKX2-5, n = 1; PAX8, n = 3; FOXE1, n = 0.

Conclusions

Conclusions. Mutations in thyroid dysgenesis genes are a rare pathology. The majority of variants among our patients were identified in TSHR.

дисгенезиявысокопроизводительное параллельное секвенированиеврожденный гипотиреозthyroid dysgenesisnext generation sequencingcongenital hypothyroidismФонд поддержки и развития филантропии “КАФ”.Charities Aid Foundation (CAF)ReferencesDevos H, Rodd C, Gagne N, et al. A search for the possible molecular mechanisms of thyroid dysgenesis: sex ratios and associated malformations. J Clin Endocrinol Metab. 1999;84(7): 2502-2506. doi: 10.1210/jcem.84.7.5831.Devos H, Rodd C, Gagne N, et al. A search for the possible molecular mechanisms of thyroid dysgenesis: sex ratios and associated malformations. J Clin Endocrinol Metab. 1999;84(7): 2502-2506. doi: 10.1210/jcem.84.7.5831.Bubuteishvili L, Garel C, Czernichow P, Léger J. Thyroid abnormalities by ultrasonography in neonates with congenital hypothyroidism. J Pediatr. 2003;143(6):759-764. doi: 10.1067/s0022-3476(03)00537-7.Bubuteishvili L, Garel C, Czernichow P, Léger J. Thyroid abnormalities by ultrasonography in neonates with congenital hypothyroidism. J Pediatr. 2003;143(6):759-764. doi: 10.1067/s0022-3476(03)00537-7.Van Vliet G, Deladoey J. Hypothyroidism in infants and children: congenital hypothyroidism. In: Braverman LE, Cooper D, editors. Werner & Ingbar’s the thyroid: a fundamental and clinical text. 10th ed. Philadelphia: Lippincott Williams and Wilkins; 2012. p. 790–802.Van Vliet G, Deladoey J. Hypothyroidism in infants and children: congenital hypothyroidism. In: Braverman LE, Cooper D, editors. Werner & Ingbar’s the thyroid: a fundamental and clinical text. 10th ed. Philadelphia: Lippincott Williams and Wilkins; 2012. p. 790–802.Rastogi MV, LaFranchi SH. Congenital hypothyroidism. Orphanet J Rare Dis. 2010;5(1):17. doi: 10.1186/1750-1172-5-17.Rastogi MV, LaFranchi SH. Congenital hypothyroidism. Orphanet J Rare Dis. 2010;5(1):17. doi: 10.1186/1750-1172-5-17.Szinnai G. Clinical genetics of congenital hypothyroidism. Endocr Dev. 2014;26:60-78. doi: 10.1159/000363156.Szinnai G. Clinical genetics of congenital hypothyroidism. Endocr Dev. 2014;26:60-78. doi: 10.1159/000363156.Lints TJ, Parsons LM, Hartley L, et al. Nkx-2.5: a novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants. Development. 1993;119(2):419-431.Lints TJ, Parsons LM, Hartley L, et al. Nkx-2.5: a novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants. Development. 1993;119(2):419-431.Dentice M, Cordeddu V, Rosica A, et al. Missense mutation in the transcription factor NKX2-5: a novel molecular event in the pathogenesis of thyroid dysgenesis. J Clin Endocrinol Metab. 2006;91(4): 1428-1433. doi: 10.1210/jc.2005-1350.Dentice M, Cordeddu V, Rosica A, et al. Missense mutation in the transcription factor NKX2-5: a novel molecular event in the pathogenesis of thyroid dysgenesis. J Clin Endocrinol Metab. 2006;91(4): 1428-1433. doi: 10.1210/jc.2005-1350.Wang K, Li M, Hakonarson H. ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res. 2010;38(16):e164. doi: 10.1093/nar/gkq603.Wang K, Li M, Hakonarson H. ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res. 2010;38(16):e164. doi: 10.1093/nar/gkq603.Richards S, Aziz N, Bale S, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015; 17(5):405-424. doi: 10.1038/gim.2015.30.Richards S, Aziz N, Bale S, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015; 17(5):405-424. doi: 10.1038/gim.2015.30.den Dunnen JT, Dalgleish R, Maglott DR, et al. HGVS Recommendations for the Description of Sequence Variants: 2016 Update. Hum Mutat. 2016;37(6):564-569. doi: 10.1002/humu.22981.den Dunnen JT, Dalgleish R, Maglott DR, et al. HGVS Recommendations for the Description of Sequence Variants: 2016 Update. Hum Mutat. 2016;37(6):564-569. doi: 10.1002/humu.22981.Sunthornthepvarakui T, Gottschalk ME, Hayashi Y, Refetoff S. Brief report: resistance to thyrotropin caused by mutations in the thyrotropin-receptor gene. N Engl J Med. 1995;332(3):155-160. doi: 10.1056/NEJM199501193320305.Sunthornthepvarakui T, Gottschalk ME, Hayashi Y, Refetoff S. Brief report: resistance to thyrotropin caused by mutations in the thyrotropin-receptor gene. N Engl J Med. 1995;332(3):155-160. doi: 10.1056/NEJM199501193320305.Макрецкая НА, Калинченко НЮ, Васильев ЕВ, и др. Клинический случай врожденного гипотиреоза, обусловленного дефектом гена NKX2-1. // Проблемы эндокринологии. – 2016. – Т. 62. – №3. – С. 21–24. [Makretskaya NA, Kalinchenko NY, Vasiliev EV, et al. Case of congenital hypothyroidism related to NKX2-1. Problems of Endocrinology. 2016;62(3):21-24. (In Russ.)] doi: 10.14341/probl201662321-24.Макрецкая НА, Калинченко НЮ, Васильев ЕВ, и др. Клинический случай врожденного гипотиреоза, обусловленного дефектом гена NKX2-1. // Проблемы эндокринологии. – 2016. – Т. 62. – №3. – С. 21–24. [Makretskaya NA, Kalinchenko NY, Vasiliev EV, et al. Case of congenital hypothyroidism related to NKX2-1. Problems of Endocrinology. 2016;62(3):21-24. (In Russ.)] doi: 10.14341/probl201662321-24.Gras D, Jonard L, Roze E, et al. Benign hereditary chorea: phenotype, prognosis, therapeutic outcome and long term follow-up in a large series with new mutations in the TITF1/NKX2-1 gene. J Neurol Neurosurg Psychiatry. 2012;83(10):956-962. doi: 10.1136/jnnp-2012-302505.Gras D, Jonard L, Roze E, et al. Benign hereditary chorea: phenotype, prognosis, therapeutic outcome and long term follow-up in a large series with new mutations in the TITF1/NKX2-1 gene. J Neurol Neurosurg Psychiatry. 2012;83(10):956-962. doi: 10.1136/jnnp-2012-302505.Montanelli L, Tonacchera M. Genetics and phenomics of hypothyroidism and thyroid dys- and agenesis due to PAX8 and TTF1 mutations. Mol Cell Endocrinol. 2010;322(1-2):64-71. doi: 10.1016/j.mce.2010.03.009.Montanelli L, Tonacchera M. Genetics and phenomics of hypothyroidism and thyroid dys- and agenesis due to PAX8 and TTF1 mutations. Mol Cell Endocrinol. 2010;322(1-2):64-71. doi: 10.1016/j.mce.2010.03.009.Narumi S, Muroya K, Abe Y, et al. TSHR mutations as a cause of congenital hypothyroidism in Japan: a population-based genetic epidemiology study. J Clin Endocrinol Metab. 2009;94(4):1317-1323. doi: 10.1210/jc.2008-1767.Narumi S, Muroya K, Abe Y, et al. TSHR mutations as a cause of congenital hypothyroidism in Japan: a population-based genetic epidemiology study. J Clin Endocrinol Metab. 2009;94(4):1317-1323. doi: 10.1210/jc.2008-1767.Al Taji E, Biebermann H, Limanova Z, et al. Screening for mutations in transcription factors in a Czech cohort of 170 patients with congenital and early-onset hypothyroidism: identification of a novel PAX8 mutation in dominantly inherited early-onset non-autoimmune hypothyroidism. Eur J Endocrinol. 2007;156(5):521-529. doi: 10.1530/EJE-06-0709.Al Taji E, Biebermann H, Limanova Z, et al. Screening for mutations in transcription factors in a Czech cohort of 170 patients with congenital and early-onset hypothyroidism: identification of a novel PAX8 mutation in dominantly inherited early-onset non-autoimmune hypothyroidism. Eur J Endocrinol. 2007;156(5):521-529. doi: 10.1530/EJE-06-0709.Bamforth JS, Hughes IA, Lazarus JH, et al. Congenital hypothyroidism, spiky hair, and cleft palate. J Med Genet. 1989;26(1):49-51. doi: 10.1136/jmg.26.1.49.Bamforth JS, Hughes IA, Lazarus JH, et al. Congenital hypothyroidism, spiky hair, and cleft palate. J Med Genet. 1989;26(1):49-51. doi: 10.1136/jmg.26.1.49.Cassio A, Nicoletti A, Rizzello A, et al. Current loss-of-function mutations in the thyrotropin receptor gene: when to investigate, clinical effects, and treatment. J Clin Res Pediatr Endocrinol. 2013;5 Suppl 1:29-39. doi: 10.4274/jcrpe.864.Cassio A, Nicoletti A, Rizzello A, et al. Current loss-of-function mutations in the thyrotropin receptor gene: when to investigate, clinical effects, and treatment. J Clin Res Pediatr Endocrinol. 2013;5 Suppl 1:29-39. doi: 10.4274/jcrpe.864.Plachov D, Chowdhury K, Walther C, et al. PAX8, a murine paired box gene expressed in the developing excretory system and thyroid gland. Development. 1990;110(2):643-651.Plachov D, Chowdhury K, Walther C, et al. PAX8, a murine paired box gene expressed in the developing excretory system and thyroid gland. Development. 1990;110(2):643-651.de Sanctis L, Corrias A, Romagnolo D, et al. Familial PAX8 small deletion (c.989_992delACCC) associated with extreme phenotype variability. J Clin Endocrinol Metab. 2004;89(11):5669-5674. doi: 10.1210/jc.2004-0398.de Sanctis L, Corrias A, Romagnolo D, et al. Familial PAX8 small deletion (c.989_992delACCC) associated with extreme phenotype variability. J Clin Endocrinol Metab. 2004;89(11):5669-5674. doi: 10.1210/jc.2004-0398.Bamforth JS, Hughes IA, Lazarus JH, et al. Congenital hypothyroidism, spiky hair, and cleft palate. J Med Genet. 1989;26(1):49-51. doi: 10.1136/jmg.26.1.49.Bamforth JS, Hughes IA, Lazarus JH, et al. Congenital hypothyroidism, spiky hair, and cleft palate. J Med Genet. 1989;26(1):49-51. doi: 10.1136/jmg.26.1.49.Clifton-Bligh RJ, Wentworth JM, Heinz P, et al. Mutation of the gene encoding human TTF-2 associated with thyroid agenesis, cleft palate and choanal atresia. Nat Genet. 1998;19(4):399-401. doi: 10.1038/1294.Clifton-Bligh RJ, Wentworth JM, Heinz P, et al. Mutation of the gene encoding human TTF-2 associated with thyroid agenesis, cleft palate and choanal atresia. Nat Genet. 1998;19(4):399-401. doi: 10.1038/1294.Stenson PD, Mort M, Ball EV, et al. The Human Gene Mutation Database: towards a comprehensive repository of inherited mutation data for medical research, genetic diagnosis and next-generation sequencing studies. Hum Genet. 2017;136(6):665-677. doi: 10.1007/s00439-017-1779-6.Stenson PD, Mort M, Ball EV, et al. The Human Gene Mutation Database: towards a comprehensive repository of inherited mutation data for medical research, genetic diagnosis and next-generation sequencing studies. Hum Genet. 2017;136(6):665-677. doi: 10.1007/s00439-017-1779-6.Lek M, Karczewski KJ, Minikel EV, et al. Analysis of protein-coding genetic variation in 60,706 humans. Nature. 2016;536(7616): 285-291. doi: 10.1038/nature19057.Lek M, Karczewski KJ, Minikel EV, et al. Analysis of protein-coding genetic variation in 60,706 humans. Nature. 2016;536(7616): 285-291. doi: 10.1038/nature19057.

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