Mutations of NKX2.5 and GATA4 genes in the development of congenital heart disease

• NKX2.5 and GATA4 mutations associated with CHD development
• NKX2.5 and GATA4 mutations affected differentiation of BMSCs into cardiomyocytes
• NKX2.5 and GATA4 affected cardiac-specific markers cTnT, Cx43, β-MHC and MLC-2
• GATA4 can interact with NKX2.5 to improve the BNP promoter activity.
• Our study provided guidance for prenatal genetic diagnosis and therapy for CHD.

ObjectiveTo investigate the mutations of NKX2.5 and GATA4 genes in the development of CHD in Chinese population.MethodsBetween December 2010 and December 2014, 185 cases of CHD patients and 210 cases of healthy people were enrolled. NKX2.5 and GATA4 gene mutations and gene expression were detected via DNA sequencing and real-time PCR (RT-PCR), respectively. BMSCs were transfected with pCMV-HA-NKX2.5 and pCMV-Myc-GATA4 plasmids. Cardiac troponin T (cTnT) and connexin 43 (Cx43) and β-myosin heavy chain (β-MHC) and myosin light chain-2 (MLC-2) expressions were detected. Co-immunoprecipitation assay was used to detect the interaction of NKX2.5 and GATA4 and luciferase to detect their effect on B-type natriuretic peptide (BNP) gene promoter.ResultsNKX2.5 and GATA4 gene mutations were found in the CHD group, but not in the normal control group, and NKX2.5 and GATA4 gene expressions were significantly lower in the case group compared with the control group (both P < 0.05). Compared to the control and empty vector groups, cTnT and Cx43 expressions were significantly higher in the pCMV-HA-NKX2.5 and pCMV-Myc-GATA4 plasmid groups; β-MHC at 1–4 weeks and MLC-2 at 2–4 weeks were higher in the pCMV-HA-NKX2.5 plasmid group; and β-MHC at 2–3 weeks and MLC-2 at 3–4 weeks were higher in the pCMV-Myc-GATA4 plasmid group (all P < 0.05). NKX2.5 and GATA4 interacted in the BMSCs and co-transfected pCMV-Myc-GATA4 and pCMV-HA-NKX2.5 significantly enhanced the fluorescence intensity of BNP.ConclusionNKX2.5 and GATA4 gene mutations might participate in the development of CHD and can promote BMSCs differentiate into cardiomyocytes.