Screening and functional analysis of GATA-6 mutations in patients with nonsyndromic conotruncal defects / 中华实用儿科临床杂志

Objective@#To explore the molecular and genetic mechanism of transcription factor GATA-6 in nonsyndromic conotruncal defect (CTD) in order to provide evidence for early prevention and inheritance consultation of CTD.@*Methods@#A total of 32 cases of patients with nonsyndromic CTD and 100 healthy individuals were enrolled in the study.A total of 7 exons and bilateral partial intron-exon boundaries of GATA-6 were amplified by means of polymerase chain reaction (PCR). The PCR products were purified and directly sequenced by using an ABI Genetic Analyzer 3100 Automatic DNA sequence equipment.The acquired GATA-6 gene sequence was compared with standard gene sequence published in National Center for Biotechnology Information database, as well as the healthy control group to observe the GATA-6 gene mutations.The mutations were introduced into pcDNA3.1(+ ) by site-directed mutagenesis PCR on the basis of pcDNA3.1(+ )-GATA-6 in order to generate the GATA6-G245R mutant constructs.Wild type GATA-6, GATA-6-G245R and atrial natriuretic factor-luciferase(ANF-luciferase) were cotransfected into HEK 293T cells in vitro, and the CMV-LacZ were cotransfected as internal reference.Luciferase and galactosidase activity were measured by using luminometer 24 h after transfection and detected in the downstream ANF-luciferase reporter gene.@*Results@#A heterozygous missense mutation in the GATA-6 gene was identified in a patient with double outlets of the right ventricle.The mutation was located in Gly245Arg(G245R) in exon 2 of GATA-6.The mutation of pcDNA3.1(+ )-GATA-6 expression vectors were successfully constructed.Through the detection of luciferase reporter gene activity, it was found that GATA-6-G245R and wild-type GATA-6 decreased by 41.3%, and the comparison between them was statistically significant (P<0.001).@*Conclusions@#Transcription factor GATA-6 gene mutation is associated with the occurrence of nonsyndromic CTD.Transcription factor GATA-6 gene may be susceptible gene in human nonsyndromic CTD.

The role of HIRA gene in the pathogenesis of congenital heart disease / 国际儿科学杂志

Conotruncal defects(CTD) is a kind of severe cardiovascular abnormality, which is the major cause of death in rinatal pelriod. Some researches suggest that CTD and 22q11 microdeletion had close relationship. In addition, HIRA gene is located in 22q11, which was speculated as a candidate gene in CTD.

Chromosome 22q11 Deletion in Patients with Infundibular Ventricular Septal Defect

PURPOSE: This study was undertaken to determine the incidence of chromosome 22q11 deletion in patients with infundibular ventricular septal defect(VSD). METHODS: Sixty-two children with infundibular VSD were included in this study from January 1999 to December 2000. Chromosome 22q11 deletion was confirmed by FISH, using LSI DiGeorge/VCFS region dual color probe(Vysis, USA). RESULTS: Thirty-two patients had conotruncal cardiac defects:tetralogy of Fallot (TOF) in 15; TOF with absent pulmonary valve in 1; VSD with pulmonary atresia in 7; truncus arteriosus in 3; double outlet right ventricle in 2; interrupted aortic arch in 2; transposition of the great arteries in 2. Thirty patients had isolated infundibular VSD without conotruncal cardiac defect:perimembranous infundibular VSD in 15; subarterial infundibular VSD in 9; muscular infundibular VSD in 6. Chromosome 22q11 deletion was observed in 8 patients(male 5, female 3):TOF 2; VSD with pulmonary atresia 4; truncus arteriosus 1; perimembranous infundibular VSD 1. All of the patients with chromosome 21q11 deletion showed typical facial appearance. Low incidence was found of chromosome 22q11 deletion in patients with infundibular VSD without conotruncal cardiac defect than in those with conotruncal cardiac defect(3.3% vs 21.9%). CONCLUSION: These data indicate that a small proportion of isolated infundibular VSD is pathogenetically related to deletion of chromosome region 22q11.

Congenital Heart Anomalies in Patients with Clefts of the Lip and/or Palate

PURPOSE: The prevalence of congenital heart anomalies is known to be higher in patients with clefts of the lip and/or palate(CL/P). The purpose of this study was to determine the prevalence and type of congenital heart anomalies in patients with CL/P. METHODS: We investigated congenital heart anomalies in 756 patients presented with CL/P from January 1986 to December 1997 by reviewing their clinical records. RESULTS: The prevalence rate of congenital heart anomalies in patients with CL/P was 4.2% (32 of 756). Congenital heart anomalies in those were ventricular septal defect (15 of 32), atrial septal defect (4 of 32), tetralogy of Fallot (3 of 32), patent ductus arteriosus (2 of 32), double outlet right ventricle(2 of 32), pulmonary stenosis (1 of 32), transposition of the great arteries (1 of 32), pulmonary atresia (1 of 32), coarctation of aorta (1 of 32), anomalous systemic venous drainage (1 of 32), and aortic aneurysm with patent ductus arteriosus (1 of 32). It was significant that the prevalence rate of congenital heart anomalies in cleft palate with or without cleft lip (CP+/-L) was 6.8% (30 of 442), because the prevalence rate of congenital heart anomalies in cleft lip alone was not higher than in normal population (0.6%; 2 of 314). Of the 30 patients with congenital heart anomalies, 12 patients (40 %) had conotruncal defects. CONCLUSION: The prevalence of congenital heart anomalies in patients with CP+/-L was much higher than normal population. Cardiac defects were predominantly conotruncal. Predominance of conotruncal defects among congenital heart anomalies in those was associated with abnormalities of neural crest cell proliferation and migration developing into conotruncus and palate.