Mutation Analysis Of CITED2 In Patients With Congenital Heart Disease

Objective:To explore gene mutation of CITED2 coding strand in different patients with CHD.To explain the relationship between CITED2 mutant and CHD phenotype providing rationale and empirical study mode for the further research on CHD.Methods:DNA was abstracted from the blood samples of 120 nonhomologous and various CHD patients and 100 healthy children. The coding regions of CITED2 was amplified by PCR and compared to the GeneBank after sequencing to identify the mutations.The family of the samples who have CITED2 mutations were investigated as well. ClustalW software was amplified for conservative analysis of the altered amino acids.Results :1. 3 new mutations of CITED2 coding strand were found in 4 CHD patients.2. 1 point mutation was first identified in a patient with mirror image dextrocardia and tetralogy of Fallot(c. 550 G>A) resulting change of CITED2 amino acid sequence(p.Gly184Ser).3. 1 point mutation was first identified in a patient with aortic stenosis(c.574 A>G)resulting change of CITED2 amino acid sequence(p. Ser192Gly).4. Apart from this, the same deletion(c.573-578del6)was first detected in other two patients,one patient with ventricular septal defect and atrial septal defect,another with aortic stenosis and pμlmonary stenosis resulting the protein changes( p. Ser192fs).5. All the changes were not detected in the control group.6. Family survey showed that the CITED2 sequence of the families with mutation cases were normal.7. Conservative analysis of the altered amino acids makes clear that the three mutant sequences have certain conservatism.Conclusions:1. This study shows for the first time that there are 3 brand-new gene mutations by sequencing of CITED2 in Chinese CHD patients with a broad phenotype spectrum. There are not obviously correlativity between mutant type and phenotype.2. All the three mutant types are in Serine-glycine rich junction (SGJ),indacating SGJ is the hot spot of CITED2 mutation. 3. Family survey shows CITED2 sequence change are the result of mutation while not heredity. Conservative analysis of the altered amino acids makes clear that the three mutant sequences have certain conservatism.4. CITED2 mutations may be one of the causative impact in the development of CHD in human.