Advertisement

Pediatric Cardiology

, Volume 40, Issue 1, pp 17–22 | Cite as

Search of Somatic Mutations of NKX2-5 and GATA4 Genes in Chinese Patients with Sporadic Congenital Heart Disease

  • Jie Yin
  • Jianhua Qian
  • Genyin Dai
  • Chunli Wang
  • Yuming Qin
  • Ting Xu
  • Zewei Li
  • Han Zhang
  • Shiwei YangEmail author
Original Article
  • 95 Downloads

Abstract

Congenital heart disease (CHD) usually occurs sporadically, with only a minority of cases associated with a known genetic mechanism. Cardiac-specific transcription factors NKX2-5 and GATA4 play key roles in the mammalian heart development, and the affected cardiac tissues of CHD patients are prone to somatic mutations which thus participate in the pathogenesis of CHD. We collected 98 patients with sporadic CHD, extracted genomic DNA from cardiac tissues and blood, and then screened NKX2-5 and GATA4 genes using PCR-direct sequence analysis. A novel heterozygous missense mutation (c.907G > A, p.V303I) of NKX2-5 gene was identified in a patient with tetralogy of Fallots. Functional assay revealed that this mutant was associated with significantly reduced transcriptional activity. In addition, we found two known single-nucleotide polymorphisms (SNPs) (rs2277923, rs3729753) in NKX2-5 and two known SNPs (rs56166237, rs3729856) in GATA4. All variations identified in cardiac tissues were consistent with those of peripheral blood, and no somatic mutations were found in cardiac tissues. Our study shows no evidence of NKX2-5 and GATA4 somatic mutations playing a role in the pathogenesis of sporadic CHD.

Keywords

Congenital heart disease Genetics Transcription factor Somatic mutation Reporter gene analysis 

Abbreviations

CHD

Congenital heart disease

TOF

Tetralogy of Fallots

ASD

Atrial septal defects

VSD

Ventricular septal defects

TAPVC

Total anomalous pulmonary venous connection

SRVOT

Stenosis of right ventricular outflow tract

DCRV

Double-chambered right ventricle

d-TGA

Transposition of the great arteries

DORV

Double outlet of right ventricle

RVOT

Right ventricular outflow tract

SNP

Single-nucleotide polymorphism.

Notes

Acknowledgements

We thank all the patients and families who participated in this study. We would like to thank all the cardiothoracic surgeons from Nanjing Children’s Hospital for their support regarding CHD heart tissues, and all the doctors of clinical laboratory.

Funding

This study was funded by the National Natural Science Foundation of China (Grant Nos. 81670284, 81000076), Nanjing Science and Technology Project (Grant No. 201715057), and Nanjing Medical Science and Technique Development Foundation.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    van der Linde D, Konings EE, Slager MA et al (2011) Birth prevalence of congenital heart disease worldwide: a systematic review and meta-analysis. J Am Coll Cardiol 58:2241–2247CrossRefGoogle Scholar
  2. 2.
    Wessels MW, Willems PJ (2010) Genetic factors in non-syndromic congenital heart malformations. Clin Genet 78:103–123CrossRefGoogle Scholar
  3. 3.
    Reamon-Buettner SM, Borlak J (2004) Somatic NKX2-5 mutations as a novel mechanism of disease in complex congenital heart disease. J Med Genet 41:684–690CrossRefGoogle Scholar
  4. 4.
    Reamon-Buettner SM, Hecker H, Spanel-Borowski K, Craatz S, Kuenzel E, Borlak J (2004) Novel NKX2-5 mutations in diseased heart tissues of patients with cardiac malformations. Am J Pathol 164:2117–2125CrossRefGoogle Scholar
  5. 5.
    Reamon-Buettner SM, Borlak J (2005) GATA4 zinc finger mutations as a molecular rationale for septation defects of the human heart. J Med Genet 42:e32CrossRefGoogle Scholar
  6. 6.
    Reamon-Buettner SM, Borlak J (2006) HEY2 mutations in malformed hearts. Hum Mutat 27:118CrossRefGoogle Scholar
  7. 7.
    Reamon-Buettner SM, Borlak J (2004) TBX5 mutations in non-Holt-Oram syndrome (HOS) malformed hearts. Hum Mutat 24:104CrossRefGoogle Scholar
  8. 8.
    Reamon-Buettner SM, Ciribilli Y, Traverso I, Kuhls B, Inga A, Borlak J (2009) A functional genetic study identifies HAND1 mutations in septation defects of the human heart. Hum Mol Genet 18:3567–3578CrossRefGoogle Scholar
  9. 9.
    Orjuela Quintero DC, Nunez F, Caicedo V, Pachon S, Salazar Salazar M (2014) Mutations in the GATA4 gen in patients with non-syndromic congenital heart disease. Invest Clin 55:207–216Google Scholar
  10. 10.
    Zheng J, Li F, Liu J et al (2015) Investigation of somatic NKX2-5 mutations in Chinese children with congenital heart disease. Int J Med Sci 12:538–543CrossRefGoogle Scholar
  11. 11.
    Sabina S, Pulignani S, Rizzo M et al (2013) Germline hereditary, somatic mutations and microRNAs targeting-SNPs in congenital heart defects. J Mol Cell Cardiol 60:84–89CrossRefGoogle Scholar
  12. 12.
    Esposito G, Butler TL, Blue GM et al (2011) Somatic mutations in NKX2-5, GATA4, and HAND1 are not a common cause of tetralogy of Fallot or hypoplastic left heart. Am J Med Genet A 155A:2416–2421CrossRefGoogle Scholar
  13. 13.
    Salazar M, Consoli F, Villegas V et al (2011) Search of somatic GATA4 and NKX2.5 gene mutations in sporadic septal heart defects. Eur J Med Genet 54:306–309CrossRefGoogle Scholar
  14. 14.
    Majumdar R, Yagubyan M, Sarkar G, Bolander ME, Sundt TM 3rd (2006) Bicuspid aortic valve and ascending aortic aneurysm are not associated with germline or somatic homeobox NKX2-5 gene polymorphism in 19 patients. J Thorac Cardiovasc Surg 131:1301–1305CrossRefGoogle Scholar
  15. 15.
    Draus JM Jr, Hauck MA, Goetsch M, Austin EH 3rd (2009) Tomita-Mitchell A and Mitchell ME: investigation of somatic NKX2-5 mutations in congenital heart disease. J Med Genet 46:115–122CrossRefGoogle Scholar
  16. 16.
    Wang J, Lu Y, Chen H, Yin M, Yu T, Fu Q (2011) Investigation of somatic NKX2-5, GATA4 and HAND1 mutations in patients with tetralogy of Fallot. Pathology 43:322–326CrossRefGoogle Scholar
  17. 17.
    Elliott DA, Kirk EP, Yeoh T et al (2003) Cardiac homeobox gene NKX2-5 mutations and congenital heart disease: associations with atrial septal defect and hypoplastic left heart syndrome. J Am Coll Cardiol 41:2072–2076CrossRefGoogle Scholar
  18. 18.
    McElhinney DB, Geiger E, Blinder J, Benson DW, Goldmuntz E (2003) NKX2.5 mutations in patients with congenital heart disease. J Am Coll Cardiol 42:1650–1655CrossRefGoogle Scholar
  19. 19.
    Nemer G, Fadlalah F, Usta J et al (2006) A novel mutation in the GATA4 gene in patients with Tetralogy of Fallot. Hum Mutat 27:293–294CrossRefGoogle Scholar
  20. 20.
    Schluterman MK, Krysiak AE, Kathiriya IS et al (2007) Screening and biochemical analysis of GATA4 sequence variations identified in patients with congenital heart disease. Am J Med Genet A 143A:817–823CrossRefGoogle Scholar
  21. 21.
    Erickson RP (2014) Recent advances in the study of somatic mosaicism and diseases other than cancer. Curr Opin Genet Dev 26:73–78CrossRefGoogle Scholar
  22. 22.
    Priest JR, Gawad C, Kahlig KM et al (2016) Early somatic mosaicism is a rare cause of long-QT syndrome. Proc Natl Acad Sci USA 113:11555–11560CrossRefGoogle Scholar
  23. 23.
    Luks VL, Kamitaki N, Vivero MP et al (2015) Lymphatic and other vascular malformative/overgrowth disorders are caused by somatic mutations in PIK3CA. J Pediatr 166:1048–1054 (e1041–1045)CrossRefGoogle Scholar
  24. 24.
    Gollob MH, Jones DL, Krahn AD et al (2006) Somatic mutations in the connexin 40 gene (GJA5) in atrial fibrillation. N Engl J Med 354:2677–2688CrossRefGoogle Scholar
  25. 25.
    Thibodeau IL, Xu J, Li Q et al (2010) Paradigm of genetic mosaicism and lone atrial fibrillation: physiological characterization of a connexin 43-deletion mutant identified from atrial tissue. Circulation 122:236–244CrossRefGoogle Scholar
  26. 26.
    Coura R, Prolla JC, Meurer L, Ashton-Prolla P (2005) An alternative protocol for DNA extraction from formalin fixed and paraffin wax embedded tissue. J Clin Pathol 58:894–895CrossRefGoogle Scholar
  27. 27.
    Quach N, Goodman MF, Shibata D (2004) In vitro mutation artifacts after formalin fixation and error prone translesion synthesis during PCR. BMC Clin Pathol 4:1CrossRefGoogle Scholar
  28. 28.
    Erickson RP (2010) Somatic gene mutation and human disease other than cancer: an update. Mutat Res 705:96–106CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of CardiologyChildren’s Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.Jiangsu Key Laboratory of PediatricsNanjing Medical UniversityNanjingChina
  3. 3.Nanjing Key Laboratory of Pediatrics, Nanjing Children’s HospitalAffiliated to Nanjing Medical UniversityNanjingChina

Personalised recommendations