DNA methylation status of NKX2-5, GATA4 and HAND1in patients with tetralogy of fallot

Abstract

BackgroundNKX2-5, GATA4 and HAND1 are essential for heart development, however, little is known regarding their epigenetic regulation in the pathogenesis of tetralogy of fallot (TOF).MethodsMethylation levels were measured in three regions of NKX2-5 (M1: -1596 bp ~ -1374 bp, M2: -159 bp ~ 217 bp and M3: 1058 bp ~ 1524 bp), one region of GATA4 (M: -392 bp ~ 107 bp) and three regions of HAND1 (M1: -887 bp ~ -414 bp, M2: -436 bp ~ 2 bp and M3: 37 bp ~ 398 bp) using the Sequenom MassARRAY platform. QRT-PCR was used to analyze NKX2-5 and HAND1 mRNA levels in the right ventricular myocardium of TOF patients.ResultsTOF patients had a significantly higher NKX2-5_M3 median methylation level than controls (41.65% vs. 22.18%; p = 0.0074; interquartile range [IQR]: 30.46%–53.35%, N = 30 and 20.07%–24.31%, N = 5; respectively). The HAND1_M1 median methylation level was also significantly higher in TOF patients than controls (30.05% vs. 17.54%; p = 0.0054; IQR: 20.77%–40.89%, N = 30 and IQR: 14.69%–20.64%; N = 6; respectively). The methylation statuses of NKX2-5_M1, NKX2-5_M2, GATA4_M, HAND1_M2 or HAND1_M3 were not significantly different in TOF patients compared to controls. The methylation values for NKX2-5_M3 were negatively correlated with mRNA levels (r = - 0.463, p = 0.010, N = 30) and there was a significant association between HAND1_M1 methylation status and mRNA levels (r = - 0.524, p = 0.003, N = 30) in TOF patients.ConclusionsAberrant methylation statuses of the NKX2-5 gene body and HAND1 promoter regions are associated with the regulation of gene transcription in TOF patients and may play an important role in the pathogenesis of TOF.

An epigenetic view of developmental diseases: new targets, new therapies

“…In sporadic tetralogy of Fallot (TOF), GATA-4, NKX2-5 and HAND1displayed aberrant DNA methylation status at the promoter region and gene body. [42] The global DNA hypermethylation has also been found in CHD children and their mothers, suggesting a role of DNA methylation in CHD. [43,44] The inactivation of Ezh2, a subunit of Polycomb repressive complex 2 (PRC2), causes lethal congenital heart malformations.…”

Existence of mutations in the homeodomain-encoding region of NKX2.5 gene in Iranian patients with tetralogy of Fallot

“…Some genes are responsible for TOF, including mutations in NKX2.5,[15222627] GATA4 interacts physically with NKX2.5,[28] GATA6,[293031] JAG1,[11323334] JAG5,[35] TBX20,[36] BVES,[37] mitochondrial ATP8 gene,[38] epigenetic changes of some genes such as NKX2.5,[39] HAND1,[39] VANGL2,[40] and single nucleotide polymorphisms of some genes such as PTPN11[41] and MTHFR. [42] In addition, TOF has been observed to be concomitant with some syndromes and associations such as Down, Alagille, DiGeorge, and CHARGE syndromes, and VACTERL association.…”

CpG island shore methylation of ZFPM2 is identified in tetralogy of fallot samples

“…The decreased LINE-1 methylation levels may be caused by the lower expression of DNMT1 and DNMT3B (21). In the present study, we focus on the DNA methylation changes within the (22) and our previous study (23). The methylation levels for the promoter region of ZFPM2 gene were initially explored in the cardiac tissues of 10 TOF patients and 6 age-matched controls, and a significant difference in the methylation levels of CpG island shore of ZFPM2 gene (ZFPM2_R1) was observed (P = 0.0256).…”