Mutation analysis of the NKX2.5 gene in Iranian pediatric patients with congenital hypothyroidism
-
Mehri Khatami
,
Mohammad Mehdi Heidari
Abstract
Background:
The embryonic development of the thyroid gland is regulated by the expression of several candidate genes which are related to congenital hypothyroidism. These genes include the numerous critical thyroid transcription factors such as NKX2.1, NKX2.5, FOXE1, and PAX8. The molecular analysis of these loci will be essential to the explanation of the participation of these transcription activators in the etiology of hypothyroidism. Among them, the role of NKX2.5 is important during the early thyroid morphogenesis and in controlling thyroidal cell differentiation and migration. Importantly, NKX2.5 change nucleotides are recognized to be central to the genesis of congenital hypothyroidism.
Methods:
A case-control study was conducted in 65 unrelated patients, diagnosed with primary congenital hypothyroidism and all of them were diagnosed according to the clinical presentations of thyroid hypoplasia and without cardiovascular defects. Mutational screening of the entire NKX2–5 coding sequence was performed in a cohort of pediatric patients by PCR-SSCP and direct sequencing.
Results:
We identified two known variations 73C>T (R25C) and 63A>G (E21E) in patients with thyroid hypothyroidism. Both of them are located in conserved region of the gene and previously reported in cases with thyroid dysgenesis and congenital heart defects. There was a significance association between 63A>G variation with primary hypothyroidism (p=0.003).
Conclusions:
These SNPs are probably related to thyroid hypoplasia because the allele frequency of the 63A>G polymorphism was significantly different in patients and controls and also R25C variation not observed in healthy cases.
Acknowledgments
We thank all the patients for providing blood samples for the scientific research.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: This research was funded by Yazd University.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Kopp P. Perspective: genetic defects in the etiology of congenital hypothyroidism. Endocrinology 2002;143:2019–24.10.1210/endo.143.6.8864Search in Google Scholar
2. Rastogi MV, LaFranchi SH. Congenital hypothyroidism. Orphanet J Rare Dis 2010;5:17.10.1186/1750-1172-5-17Search in Google Scholar
3. van Engelen K, Mommersteeg MT, Baars MJ, Lam J, Ilgun A, et al. The ambiguous role of NKX2-5 mutations in thyroid dysgenesis. PLoS One 2012;7:e52685.10.1371/journal.pone.0052685Search in Google Scholar
4. Dentice M, Cordeddu V, Rosica A, Ferrara AM, Santarpia L, 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;91:1428–33.10.1210/jc.2005-1350Search in Google Scholar
5. Castanet M, Polak M, Bonaiti-Pellie C, Lyonnet S, Czernichow P, et al. Nineteen years of national screening for congenital hypothyroidism: familial cases with thyroid dysgenesis suggest the involvement of genetic factors. J Clin Endocrinol Metab 2001;86:2009–14.10.1210/jcem.86.5.7501Search in Google Scholar
6. Bubuteishvili L, Garel C, Czernichow P, Leger J. Thyroid abnormalities by ultrasonography in neonates with congenital hypothyroidism. J Pediatr 2003;143:759–64.10.1067/S0022-3476(03)00537-7Search in Google Scholar
7. Deladoey J, Vassart G, Van Vliet G. Possible non-Mendelian mechanisms of thyroid dysgenesis. Endocr Dev 2007;10:29–42.10.1159/000106818Search in Google Scholar PubMed
8. Montanelli L, Tonacchera M. Genetics and phenomics of hypothyroidism and thyroid dys- and agenesis due to PAX8 and TTF1 mutations. Mol Cell Endocrinol 2010;322:64–71.10.1016/j.mce.2010.03.009Search in Google Scholar PubMed
9. Tong YF. Mutations of NKX2.5 and GATA4 genes in the development of congenital heart disease. Gene 2016;588:86–94.10.1016/j.gene.2016.04.061Search in Google Scholar PubMed
10. Zhao L, Ni SH, Liu XY, Wei D, Yuan F, et al. Prevalence and spectrum of Nkx2.6 mutations in patients with congenital heart disease. Eur J Med Genet 2014;57:579–86.10.1016/j.ejmg.2014.08.005Search in Google Scholar PubMed
11. Narumi S, Muroya K, Asakura Y, Adachi M, Hasegawa T. Transcription factor mutations and congenital hypothyroidism: systematic genetic screening of a population-based cohort of Japanese patients. J Clin Endocrinol Metab 2010;95:1981–5.10.1210/jc.2009-2373Search in Google Scholar PubMed
12. McElhinney DB, Geiger E, Blinder J, Benson DW, Goldmuntz E. NKX2.5 mutations in patients with congenital heart disease. J Am Coll Cardiol 2003;42:1650–5.10.1016/j.accreview.2003.12.072Search in Google Scholar
13. Jay PY, Harris BS, Maguire CT, Buerger A, Wakimoto H, et al. Nkx2-5 mutation causes anatomic hypoplasia of the cardiac conduction system. J Clin Invest 2004;113:1130–7.10.1172/JCI19846Search in Google Scholar PubMed PubMed Central
14. Biben C, Weber R, Kesteven S, Stanley E, McDonald L, et al. Cardiac septal and valvular dysmorphogenesis in mice heterozygous for mutations in the homeobox gene Nkx2-5. Circ Res 2000;87:888–95.10.1161/01.RES.87.10.888Search in Google Scholar PubMed
15. Kurian MA, Jungbluth H. Genetic disorders of thyroid metabolism and brain development. Dev Med Child Neurol 2014;56:627–34.10.1111/dmcn.12445Search in Google Scholar PubMed PubMed Central
16. De Felice M, Di Lauro R. Thyroid development and its disorders: genetics and molecular mechanisms. Endocr Rev 2004;25: 722–46.10.1210/er.2003-0028Search in Google Scholar PubMed
17. Vincentz JW, Barnes RM, Firulli BA, Conway SJ, Firulli AB. Cooperative interaction of Nkx2.5 and Mef2c transcription factors during heart development. Dev Dyn 2008;237:3809–19.10.1002/dvdy.21803Search in Google Scholar PubMed PubMed Central
18. Akazawa H, Komuro I. Cardiac transcription factor Csx/Nkx2-5: its role in cardiac development and diseases. Pharmacol Ther 2005;107:252–68.10.1016/j.pharmthera.2005.03.005Search in Google Scholar PubMed
19. Beffagna G, Cecchetto A, Dal Bianco L, Lorenzon A, Angelini A, et al. R25C mutation in the NKX2.5 gene in Italian patients affected with non-syndromic and syndromic congenital heart disease. J Cardiovasc Med (Hagerstown) 2013;14:582–6.10.2459/JCM.0b013e328356a326Search in Google Scholar PubMed
20. Cerqueira TL, Ramos Y, Strappa G, Martin DS, Jesus M, et al. The c.63A>G polymorphism in the NKX2.5 gene is associated with thyroid hypoplasia in children with thyroid dysgenesis. Arch Endocrinol Metab 2015;59:562–7.10.1590/2359-3997000000100Search in Google Scholar PubMed
21. Draus JM, Jr., Hauck MA, Goetsch M, Austin EH, 3rd, Tomita-Mitchell A, et al. Investigation of somatic NKX2-5 mutations in congenital heart disease. J Med Genet 2009;46:115–22.10.1136/jmg.2008.060277Search in Google Scholar
22. Reamon-Buettner SM, Hecker H, Spanel-Borowski K, Craatz S, Kuenzel E, et al. Novel NKX2-5 mutations in diseased heart tissues of patients with cardiac malformations. Am J Pathol 2004;164:2117–25.10.1016/S0002-9440(10)63770-4Search in Google Scholar
23. Inga A, Reamon-Buettner SM, Borlak J, Resnick MA. Functional dissection of sequence-specific NKX2-5 DNA binding domain mutations associated with human heart septation defects using a yeast-based system. Hum Mol Genet 2005;14:1965–75.10.1093/hmg/ddi202Search in Google Scholar PubMed
24. Fagman H, Andersson L, Nilsson M. The developing mouse thyroid: embryonic vessel contacts and parenchymal growth pattern during specification, budding, migration, and lobulation. Dev Dyn 2006;235:444–55.10.1002/dvdy.20653Search in Google Scholar PubMed
25. Ramos HE, Nesi-Franca S, Boldarine VT, Pereira RM, Chiamolera MI, et al. Clinical and molecular analysis of thyroid hypoplasia: a population-based approach in southern Brazil. Thyroid 2009;19:61–8.10.1089/thy.2008.0116Search in Google Scholar PubMed
26. Al Taji E, Biebermann H, Limanova Z, Hnikova O, Zikmund J, 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;156:521–9.10.1530/EJE-06-0709Search in Google Scholar PubMed
27. Brust ES, Beltrao CB, Chammas MC, Watanabe T, Sapienza MT, et al. Absence of mutations in PAX8, NKX2.5, and TSH receptor genes in patients with thyroid dysgenesis. Arq Bras Endocrinol Metabol 2012;56:173–7.10.1590/S0004-27302012000300004Search in Google Scholar
©2017 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
- Frontmatter
- Editorial
- Endocrine aspects in cystic fibrosis
- Original Articles
- A retrospective analysis of longitudinal changes in bone mineral content in cystic fibrosis
- Cystic-fibrosis related-diabetes (CFRD) is preceded by and associated with growth failure and deteriorating lung function
- Partial clinical remission in type 1 diabetes: a comparison of the accuracy of total daily dose of insulin of <0.3 units/kg/day to the gold standard insulin-dose adjusted hemoglobin A1c of ≤9 for the detection of partial clinical remission
- Concentrations of leptin, adiponectin and other metabolic parameters in non-obese children with Down syndrome
- Parent reported nutritional risk and laboratory indices of cardiometabolic risk and in preschool-aged children
- Multinodular goiter in children: treatment controversies
- Atopy as a risk factor for subclinical hypothyroidism development in children
- Mutation analysis of the NKX2.5 gene in Iranian pediatric patients with congenital hypothyroidism
- Health-related quality of life among children with Turner syndrome: controlled cross-sectional study
- Growth and pubertal patterns in young survivors of childhood acute lymphoblastic leukemia
- Clinical features and genotyping of patients with primary carnitine deficiency identified by newborn screening
- Letter to the Editor
- Sensitivity and specificity of cystic fibrosis-related diabetes screening methods: which test should be the reference method?
- Case Reports
- Type 1 rhizomelic chondrodysplasia punctata with a homozygous PEX7 mutation
- Severe hypertriglyceridemia at new onset type 1 diabetes mellitus
- 45,X/46,XY ovotesticular disorder of sex development revisited: undifferentiated gonadal tissue may be mistaken as ovarian tissue
- MRI in medium-chain acyl-coenzyme a dehydrogenase deficiency: neuroimaging during the first month
Articles in the same Issue
- Frontmatter
- Editorial
- Endocrine aspects in cystic fibrosis
- Original Articles
- A retrospective analysis of longitudinal changes in bone mineral content in cystic fibrosis
- Cystic-fibrosis related-diabetes (CFRD) is preceded by and associated with growth failure and deteriorating lung function
- Partial clinical remission in type 1 diabetes: a comparison of the accuracy of total daily dose of insulin of <0.3 units/kg/day to the gold standard insulin-dose adjusted hemoglobin A1c of ≤9 for the detection of partial clinical remission
- Concentrations of leptin, adiponectin and other metabolic parameters in non-obese children with Down syndrome
- Parent reported nutritional risk and laboratory indices of cardiometabolic risk and in preschool-aged children
- Multinodular goiter in children: treatment controversies
- Atopy as a risk factor for subclinical hypothyroidism development in children
- Mutation analysis of the NKX2.5 gene in Iranian pediatric patients with congenital hypothyroidism
- Health-related quality of life among children with Turner syndrome: controlled cross-sectional study
- Growth and pubertal patterns in young survivors of childhood acute lymphoblastic leukemia
- Clinical features and genotyping of patients with primary carnitine deficiency identified by newborn screening
- Letter to the Editor
- Sensitivity and specificity of cystic fibrosis-related diabetes screening methods: which test should be the reference method?
- Case Reports
- Type 1 rhizomelic chondrodysplasia punctata with a homozygous PEX7 mutation
- Severe hypertriglyceridemia at new onset type 1 diabetes mellitus
- 45,X/46,XY ovotesticular disorder of sex development revisited: undifferentiated gonadal tissue may be mistaken as ovarian tissue
- MRI in medium-chain acyl-coenzyme a dehydrogenase deficiency: neuroimaging during the first month
