Pathogenic Gene Research Based On Cases Diagnosed With Isolated Coarctation Of The Aorta

Object:Aiming to find the pathogenic gene,venous blood DNA whole-exome seq uencing has been done in one core family including two cases of isolated coar ctation of the aorta.It will contribute to deepen the understanding of the path ogenesis of aortic coarctation.Method:Patients are a pair of monozygotic twins,diagnosed with coarctation of th e aorta by aortic computerized tomography angiography.Patients did not have family history of congenital heart diseases or connective tissue diseases.Contro ls are patients' parents and brother.Venous blood DNA whole-exome sequencin g was done using illumina hiseq 2500.Sequencing methods adopt paired-end s equencing with cover depth more than 100 X on average.Data processing inclu des sequence alignment removing possible PCR repetition calling SNVs(s ingle-nucleotide variants)and In Dels(insertion-deletion).Preliminary results wer e obtained depending on mutation location and mutation type.Candidate results were obtained following de onovo mutation autosomal recessive mutation an d compound heterozygous mutation.Sanger sequencing was utilized to verify mutations.In the end,Bioinformatic analysis was done to acquire potential pat hogenic gene.Genetic conservation detrimental mutations Pathway analysis Go analysis domain function expression and publication were evaluated in the bioinformatic analysis.Result:we found 23643 SNVs and 507 In Dels based on venous blood DNA whole-exome sequencing of one core family including two cases.We keep the mutat ions judged with pass by software.After retaining mutations located in exonic and splicing,there were 23643 SNVs and 507 In Del.After keeping mutations having impact on amino acid,there were 10762 SNVs and 121 In Dels.There were not In Dels following inheritance patterns based on core family.However,there were 11 SNVs meeting the conditions.AQP12 c.T83G:p.L28 R and F BXO7 c.G212A:p.G71E/c.G212C:p.G71 A meet de novo mutation.SAMD9 L c.T866C:p.F289 S CCDC132 c.A1G:p.M1 V LAMB4 c.C700T:p.H234 Y and CHPT1 c.T1021C:p.F341 L fit autosomal recessive mutations.DNAH8 c.G7171T:p.A2391S/c.C14075T:p.P4692 L FAM194A c.G1055T:p.R352L/c.C472T:p.R158 C FAT2 c.G5647A:p.G1883R/c.G2975A:p.R992 Q GGA c.G1480A:p.E494 K c.G1747A:p.E583 K c.G1630A:p.E544 K c.G1846A:p.E616K/c.G289C:p.E97 Q c.G556C:p.E186 Q c.G439C:p.E147 Q c.G655C:p.E219 Q and SSPO c.G1703A:p.G568E/c.C2882T:p.T961 M conform compound heterozygous mut ations.Excluding AQP12 c.T83G:p.L28 R,all the genes passed validation in sanger sequencing.In the end,potential pathogenic genes were obtained via bi oinformatic analysis: gene SSPO,possibly associated with abnormal neural cres t cell development,contributing to abnormal development of the aortic wall.Conclusion:Gene SSPO maybe potential pathogenic gene contributing to isolated coarc tation of the aorta as a rare mutation.It is not reported before our research.C omplete physical examination and related auxiliary examination should be done in the patient who carries c.G1703A:p.G568 E and c.C2882T:p.T961 M at the same time to confirm whether the patient suffer from aortic coarctation.If the parents carry one of two mutations respectively,there is enhanced possibility t o develop into aortic coarctation in their offspring.