新疆维吾尔族非综合征型耳聋家系致病基因研究

When studying on Xinjiang Uyghur deaf people at early stage, we find the common deafness gene of over 20 proposituses with family history is detected to be negative and collect and sort the deaf family information. This project intends to select 10 non-syndromic deaf families for pathogenic gene research and have preliminary screening on the commonest mutation site of deafness gene in family members by deafness gene chip; the family without mutation after inspection is further subject to full sequencing test of hot-site deafness gene GJB2; then, for the family with test result of being negative, the DNA sequence of exon area in the whole genome is captured by high-throughput exon sequencing technology in order to sequence concentratedly and analyze and seek the mutation site, thereby finding the candidate genes, finally determine the deafness pathogenic gene of the family and expect to find new mutation gene or site. This study is favorable for further determining the hot-site mutation of Uyghur nationality, providing theoretical basis for developing screening and preventive strategy of Uyghur deaf people and establishing basis for deafness gene function and individual treatment study later.

我们在前期对新疆耳聋人群的研究中，发现维吾尔族耳聋人群常见耳聋基因突变检出率低，收集的20余个维吾尔族耳聋家系，其先证者常见耳聋基因突变筛查为阴性。本项目拟选取其中10个非综合征型耳聋小家系进行致病基因研究，利用耳聋基因芯片初步筛查家系成员常见的耳聋基因突变，未检测出突变的家系进一步行耳聋热点基因GJB2全序列测序，测序结果为阴性的家系，然后应用高通量外显子测序技术，将全基因组外显子区域的DNA序列捕获下来，进行集中测序分析寻找突变位点，从而找到候选致病基因，最终确定家系耳聋致病基因。本研究有助于进一步丰富维吾尔族耳聋人群热点基因与热点突变，为下一步制定更适合新疆维吾尔族耳聋人群的耳聋基因筛查方案和预防策略提供理论依据，并为后期耳聋基因功能与个体性治疗研究奠定基础。

耳聋是耳鼻咽喉科的常见疾病，约60%的先天性耳聋是由遗传因素导致的，基因突变是导致遗传性耳聋的原因，前期研究表明新疆维吾尔族耳聋热点基因和热点突变与国内耳聋人群不完全一致，有一些维吾尔族耳聋患者的致病原因仍然不明确。 .耳聋家系为遗传性耳聋致病基因的研究提供了有利资源，此课题选取J01、J02、J03、J04、J05、J06、J07、J08、J09、J10、J11、J12、J13、J14、J15、J16和J17共17个遗传性耳聋家系进行突变基因的研究，期望明确耳聋的致病基因和在人群中的发病情况。.研究先利用天昊耳聋基因芯片筛查家系成员最常见的GJB2 、12SrRNA和SLC26A4三个耳聋基因突变位点，筛查到以下突变，分别为：J01 mtDNA基因的 1555A>G(M/M)突变，J04 SLC26A4：1174A>T突变，J06 GJB2 235delC/257C>G突变，J12 SLC26A4基因170C>A（M/M）突变，J14 SLC26A4基因2027T>A（M/M）突变；对于检测结果为阴性的家系，进入高通量测序，通过高通量测序，检测到以下突变，J02家系MYO15A:6892C>T突变，J03家系MYO7A:1258A>T突变，J07家系MYO15A:c.9514C>T/7894G>T突变，J08家系PCDH15基因4658delT突变，J09家系TMC1:773G>A突变，J10家系GIPC3:788C>A，J11家系TMC1:1247T>G/1312G>A突变，J13家系PCDH15：4658delT突变，J16家系 MYO15A:9514C>T突变。本研究中，J17家系在补样验证时失访，J05和J15家系未检测到有意义的突变。此研究明确了J01、J02、J03、J04、J06、J07、J08、J09、J10、J11、J12、J13、J14和J16耳聋家系的耳聋致病基因和位点，其中6例是新发现突变，分别为MYO15A基因6892C>T和 9514C>T突变，MYO7A基因1258A>T，PCDH15基因4658delT突变，TMC1基因773G>A突变。.本研究明确了维吾尔族耳聋家系的耳聋原因，丰富了维吾尔族耳聋基因谱，为将来维吾尔族耳聋基因临床筛查提供了有意义的参考价值，并为后期耳聋基因功能和个体性治疗研究奠定基础。