{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T23:44:28Z","timestamp":1788219868923,"version":"build-2803163510"},"reference-count":32,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2018,6,19]],"date-time":"2018-06-19T00:00:00Z","timestamp":1529366400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Medicina"],"abstract":"<jats:p>Background and objective: Congenital heart disease (CHD) is the most common birth abnormality in the structure or function of the heart that affects approximately 1% of all newborns. Despite its prevalence and clinical importance, the etiology of CHD remains mainly unknown. Somatic and germline mutations in cardiac specific transcription factor genes have been identified as the factors responsible for various forms of CHD, particularly ventricular septal defects (VSDs), tetralogy of Fallot (TOF), and atrial septal defects (ASDs). p. NKX2.5 is a homeodomain protein that controls many of the physiological processes in cardiac development including specification and proliferation of cardiac precursors. The aim of our study was to evaluate the NKX2.5 gene mutations in sporadic pediatric patients with clinical diagnosis of congenital heart malformations. Materials and methods: In this study, we investigated mutations of the NKX2.5 gene\u2019s coding region in 105 Iranian pediatric patients with non-familial CHD by polymerase chain reaction-single stranded conformation polymorphism (PCR-SSCP) and direct sequencing. Results: We observed a total of four mutations, of which, two were novel DNA sequence variants in the coding region of exon 1 (c. 95 A &gt; T and c. 93 A &gt; T) and two others were previously reported as single-nucleotide polymorphisms (SNPs), namely rs72554028 (c. 2357 G &gt; A) and rs3729753 (c. 606 G &gt; C) in exon 2. Further, observed mutations are completely absent in normal healthy individuals (n = 92). Conclusion: These results suggest that NKX2.5 mutations are highly rare in CHD patients. However, in silico analysis proves that c.95 A &gt; T missense mutation in NKX2.5 gene is probably pathogenic and may be contributing to the risk of sporadic CHD in the Iranian population.<\/jats:p>","DOI":"10.3390\/medicina54030046","type":"journal-article","created":{"date-parts":[[2018,6,19]],"date-time":"2018-06-19T11:20:36Z","timestamp":1529407236000},"page":"46","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Novel Point Mutations in the NKX2.5 Gene in Pediatric Patients with Non-Familial Congenital Heart Disease"],"prefix":"10.3390","volume":"54","author":[{"given":"Mehri","family":"Khatami","sequence":"first","affiliation":[{"name":"Department of Biology, Faculty of Science, Yazd University, Yazd 8915818411, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mansoureh","family":"Mazidi","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Yazd University, Yazd 8915818411, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shabnam","family":"Taher","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Yazd University, Yazd 8915818411, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Mehdi","family":"Heidari","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Yazd University, Yazd 8915818411, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehdi","family":"Hadadzadeh","sequence":"additional","affiliation":[{"name":"Department of Cardiac Surgery, Afshar Hospital, Shahid Sadoughi University of Medical Sciences, Yazd 8915818411, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,6,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e183","DOI":"10.1016\/j.carpath.2009.06.008","article-title":"Molecular mechanisms of congenital heart disease","volume":"19","author":"Huang","year":"2010","journal-title":"Cardiovasc. Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1007\/s00018-005-5532-2","article-title":"Insights into the genetic basis of congenital heart disease","volume":"63","author":"Garg","year":"2006","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s10528-016-9780-7","article-title":"Genetics of Congenital Heart Disease: Past and Present","volume":"55","author":"Muntean","year":"2017","journal-title":"Biochem. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1053\/j.semtcvs.2007.07.013","article-title":"The molecular basis of congenital heart disease","volume":"19","author":"Mitchell","year":"2007","journal-title":"Semin. Thorac. Cardiovasc. Surg."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Chung, I.M., and Rajakumar, G. (2016). Genetics of Congenital Heart Defects: The NKX2-5 Gene, a Key Player. Genes, 7.","DOI":"10.3390\/genes7020006"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.ijcard.2008.08.035","article-title":"NKX2.5 mutations in patients with non-syndromic congenital heart disease","volume":"138","author":"Pereira","year":"2010","journal-title":"Int. J. Cardiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1515\/jpem-2017-0084","article-title":"Mutation analysis of the NKX2.5 gene in Iranian pediatric patients with congenital hypothyroidism","volume":"30","author":"Khatami","year":"2017","journal-title":"J. Pediatr. Endocrinol. Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1126\/science.281.5373.108","article-title":"Congenital heart disease caused by mutations in the transcription factor NKX2-5","volume":"281","author":"Schott","year":"1998","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1089\/gtmb.2015.0112","article-title":"Investigation of NKX2.5 gene mutations in congenital heart defects in an Indian population","volume":"19","author":"Ketharnathan","year":"2015","journal-title":"Genet. Test Mol. Biomark."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2389","DOI":"10.1016\/j.jacc.2016.08.064","article-title":"Cardiac Phenotype and Long-Term Follow-Up of Patients With Mutations in NKX2-5 Gene","volume":"68","author":"Maury","year":"2016","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1016\/j.cell.2007.01.042","article-title":"An Nkx2-5\/Bmp2\/Smad1 negative feedback loop controls heart progenitor specification and proliferation","volume":"128","author":"Prall","year":"2007","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4187","DOI":"10.1242\/dev.126.19.4187","article-title":"Building the heart piece by piece: Modularity of cis-elements regulating Nkx2-5 transcription","volume":"126","author":"Schwartz","year":"1999","journal-title":"Development"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1006\/geno.1995.1027","article-title":"Assignment of cardiac homeobox gene CSX to human chromosome 5q34","volume":"27","author":"Shiojima","year":"1995","journal-title":"Genomics"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1111\/j.1399-0004.2010.01422.x","article-title":"Mutational spectrum in the cardiac transcription factor gene NKX2.5 (CSX) associated with congenital heart disease","volume":"78","author":"Stallmeyer","year":"2010","journal-title":"Clin. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.ccep.2017.02.004","article-title":"Ventricular Arrhythmias in Adult Congenital Heart Disease: Mechanisms, Diagnosis, and Clinical Aspects","volume":"9","author":"Sathananthan","year":"2017","journal-title":"Card. Electrophysiol. Clin."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2117","DOI":"10.1016\/S0002-9440(10)63770-4","article-title":"Novel NKX2-5 mutations in diseased heart tissues of patients with cardiac malformations","volume":"164","author":"Hecker","year":"2004","journal-title":"Am. J. Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1136\/jmg.2008.060277","article-title":"Investigation of somatic NKX2-5 mutations in congenital heart disease","volume":"46","author":"Draus","year":"2009","journal-title":"J. Med. Genet."},{"key":"ref_18","first-page":"47","article-title":"Phenotypes with GATA4 or NKX2.5 mutations in familial atrial septal defect","volume":"135","author":"Kamisago","year":"2005","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1016\/j.jacc.2003.05.004","article-title":"NKX2.5 mutations in patients with congenital heart disease","volume":"42","author":"McElhinney","year":"2003","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1111\/j.1747-0803.2011.00573.x","article-title":"Combined mutation screening of NKX2-5, GATA4, and TBX5 in congenital heart disease: Multiple heterozygosity and novel mutations","volume":"7","author":"Pope","year":"2012","journal-title":"Congenit. Heart Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1016\/j.gene.2013.04.043","article-title":"Two novel and functional DNA sequence variants within an upstream enhancer of the human NKX2-5 gene in ventricular septal defects","volume":"524","author":"Huang","year":"2013","journal-title":"Gene"},{"key":"ref_22","first-page":"29","article-title":"Nkx2-5 Mutations in Patients with Non-Syndrome Congenital Heart Disease","volume":"15","author":"Soheili","year":"2015","journal-title":"Zahedan J. Res. Med. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"24","DOI":"10.4103\/1735-1995.179893","article-title":"Existence of mutations in the homeodomain-encoding region of NKX2.5 gene in Iranian patients with tetralogy of Fallot","volume":"21","author":"Kheirollahi","year":"2016","journal-title":"J. Res. Med. Sci."},{"key":"ref_24","first-page":"250","article-title":"Gene mutation in secundum atrial septal defect: Analysis of a Chinese family with 3 patients","volume":"88","author":"Tian","year":"2008","journal-title":"Zhonghua Yi Xue Za Zhi"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1172\/JCI9860","article-title":"Loss of function and inhibitory effects of human CSX\/NKX2.5 homeoprotein mutations associated with congenital heart disease","volume":"106","author":"Kasahara","year":"2000","journal-title":"J. Clin. Investig."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2565","DOI":"10.1161\/hc4601.098427","article-title":"NKX2.5 mutations in patients with tetralogy of fallot","volume":"104","author":"Goldmuntz","year":"2001","journal-title":"Circulation"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Reamon-Buettner, S.M., Sattlegger, E., Ciribilli, Y., Inga, A., Wessel, A., and Borlak, J. (2013). Transcriptional defect of an inherited NKX2-5 haplotype comprising a SNP, a nonsynonymous and a synonymous mutation, associated with human congenital heart disease. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0083295"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6307","DOI":"10.1073\/pnas.96.11.6307","article-title":"A single nucleotide in the SMN gene regulates splicing and is responsible for spinal muscular atrophy","volume":"96","author":"Lorson","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1038\/83762","article-title":"A mechanism for exon skipping caused by nonsense or missense mutations in BRCA1 and other genes","volume":"27","author":"Liu","year":"2001","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1016\/j.clineuro.2009.04.004","article-title":"Mutations in the NKX2-5 gene in patients with stroke and patent foramen ovale","volume":"111","author":"Tizzano","year":"2009","journal-title":"Clin. Neurol. Neurosurg."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.gene.2015.08.033","article-title":"Genetic variations of NKX2-5 in sporadic atrial septal defect and ventricular septal defect in Chinese Yunnan population","volume":"575","author":"Cao","year":"2016","journal-title":"Gene"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1089\/gtmb.2008.0044","article-title":"Screening NKX2.5 mutation in a sample of 230 Han Chinese children with congenital heart diseases","volume":"13","author":"Zhang","year":"2009","journal-title":"Genet. Test Mol. Biomark."}],"container-title":["Medicina"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1648-9144\/54\/3\/46\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:09:14Z","timestamp":1760195354000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1648-9144\/54\/3\/46"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,6,19]]},"references-count":32,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2018,7]]}},"alternative-id":["medicina54030046"],"URL":"https:\/\/doi.org\/10.3390\/medicina54030046","relation":{},"ISSN":["1648-9144"],"issn-type":[{"value":"1648-9144","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,6,19]]}}}