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Genetic and Functional Analysis of the NKX2-5 Gene Promoter in Patients With Ventricular Septal Defects

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Abstract

The ventricular septal defect (VSD) is the most common type of congenital heart disease (CHD). The morbidity and mortality of CHD patients are significantly higher due to late cardiac complications, likely caused by genetic defects. Mutations in cardiac transcription factor genes such as GATA-4, TBX5, and NKX2-5 have been implicated in CHD cases. The NKX2-5 gene, a homeobox gene, is expressed in the developing heart and the adult heart. Because NKX2-5 is a dosage-sensitive regulator during embryonic development, the authors hypothesized that the expression levels of the NKX2-5 gene rather than the mutant protein may play important roles in CHD. In this study, the promoter regions and exon regions of the NKX2-5 gene were bidirectionally sequenced in large cohorts of VSD patients and healthy control subjects. The results showed that a novel sequence variant (g.4574c>deletion), found only in one VSD patient, and a single nucleotide polymorphism (rs118026695), the frequency of which was significantly higher in VSD patients, were identified within the promoter region. Functional analysis confirmed that these sequence variants significantly enhanced the transcriptional activities of the NKX2-5 gene promoter, altering the expression of the NKX2-5 gene and the cardiac gene regulatory network. In addition, a synonymous mutation in the second exon of the NKX2-5 gene was identified in one VSD patient, which may affect the translation process. Therefore, the authors’ data provide supportive evidence that mutations in the coding region of the NKX2-5 gene and sequence variants within its promoter region may be among the contributors to the CHD etiology.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 81070173) and the Shandong Provincial Natural Science Foundation (ZR2010HL030 and ZR2010HM111).

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Cardiac Disease Diagnosis and Treatment, Jining Medical University Affiliated Hospital, Jining Medical University, 79 Guhuai Road, Jining, 272029, Shandong, People’s Republic of China

    Shuchao Pang, Jiping Shan, Yanli Qiao, Liming Ma, Xianyun Qin, Hongxin Wanyan, Guanghua Wu & Bo Yan

  2. Division of Pediatric Surgery, Jining Medical University Affiliated Hospital, Jining Medical University, Jining, 272029, Shandong, China

    Qining Xing

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  1. Shuchao Pang
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  2. Jiping Shan
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Correspondence to Bo Yan.

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Shuchao Pang, Jiping Shan, and Yanli Qiao have contributed equally to this work.

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Pang, S., Shan, J., Qiao, Y. et al. Genetic and Functional Analysis of the NKX2-5 Gene Promoter in Patients With Ventricular Septal Defects. Pediatr Cardiol 33, 1355–1361 (2012). https://doi.org/10.1007/s00246-012-0346-0

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  • DOI: https://doi.org/10.1007/s00246-012-0346-0

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