Targeted Sequencing Of 307 Genes Associated With Hereditary Deafness In Hemifacial Microsomia Patients Complicated With Or Without Cervical Deformity

Objective:To determine whether mutations in genes associated with hereditary deafness cause microtia.Methods:1)Thirty-two microtia patients were selected and 2 ml of fasting venous blood was collected from each patient.Genomic DNA was extracted by BioChain whole blood genomic DNA extraction kit and quantified and tested by agarose gel electrophoresis.Samples were selected according to the requirements to prepare the DNA library.2)Selected the hereditary deafness genes of BGI and utilized targeted genomic capturing combined with next-generation sequencing to screen for mutations of 307 genes associated with hereditary deafness in 32 microtia patients.Related tools,softwares and methods in bioinformatics were applied in sequence alignment,data collection and variation analysis;3)Patients were classified according to clinical phenotypes and then acquire the genetic of each kind of microtia.Results:We found 37 heterozygous mutations of 28 genes in 20 patients were most associated with microtia patients.Among these,14 of 24 isolated unilateral microtia patients were identified to carry 22 mutations in GPR98,DIAPH1,FGFR2,HOXA1,EDNRB,MY07A,TRIOBP,USH1C,MYO1A,LRP2,GPSM2,COL11A1,LHFPL5,OTOG,MY03A,OPA1,WFS1 and DIAPH3 genes,4 of 5 isolated bilateral microtia patients were identified to carry 12 mutations in ALMS1,STRC,BSND,COL4A4,GPR98,LOXHD1,OTOG,COL4A5,USH2A,OTOF and PCDH15 genes and 2 of 3 isolated unilateral microtia patients complicated with hemifacial microsomia were identified to carry 3 mutations in PCDH1,COL11A1 and EYA1 genes.Mutations in LRP2,MY07A,GPR98,LOXHD1,PCDH15,WFS1,OPA1,OTOG and COL11A1 genes were relatively more frequent(≥2 patients).Conclusion:Some hereditary deafness genes may play a critical role in the pahtogenesis of microtia.Different type of microtia may have different genetic factors.To explore the genetic of microtia according to different kind of microtia patients may help to identify susceptibility genes.