USH1C基因内含子新突变导致一耳聋家系的研究  

Study on a deaf family caused by new USHiC gene intron mutation

作　　者：赵涛涛 马秀丽 林垦 明澄 马静[1] ZHAO Taotao;MA Xiuli;LIN Ken;MING Cheng;MA Jing(Department of Otolaryngology Head and Neck Surgery,Kunming Children's Hospital,Key Laboratory of Prevention and Control of Child Birth Defects in Kunming,Kunming,Yunnan,650228,China)

机构地区：[1]昆明市儿童医院耳鼻咽喉头颈外科,昆明市儿童先天出生缺陷防控研究重点实验室,云南昆明650228

出　　处：《中国耳鼻咽喉头颈外科》2023年第5期300-304,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：云南省王海波专家工作站(202105AF150056);昆明市儿童先天出生缺陷防控重点实验室(2019-1-A-24465);云南省中青年学术和技术带头人后备人才培养项目(2019HB102);云南省高层次卫生健康技术人才培养专项(L-2019002);昆明医科大学2022年研究生创新基金(2022S347)。

摘　　要：目的 对一个耳聋家系,应用高通量测序技术、Minigene实验,对其听力损失病因及分子生物学致病原因进行探究。方法 首先采集先证者及家系成员的病史、绘制家系图,并进行听力学评估及耳部影像学检查。获取该家系2代3人外周血提取基因组DNA,对患儿采用高通量测序方法对406个耳聋基因编码区的全部外显子及部分内含子和启动子进行检测,筛选疑似致病突变,针对变异位点对家系成员进行Sanger测序验证,应用Minigene实验进行验证。结果 患儿表现为双侧极重度聋,高通量测序结果为USH1C纯合突变c.388-1G>A,导致氨基酸发生剪接突变,根据美国医学遗传学与基因组学学会(ACMG)指南,该变异初步判定为疑似致病性变异;Sanger测序验证其父母均为该位点的杂合携带者。Minigene实验表明该位点突变会导致m RNA异常剪接。结论 本家系中发现的USH1C基因内含子新突变为致病性突变,该突变导致先证者耳聋。OBJECTIVE To explore the causes of hearing loss and molecular biology in a family with deafness using high-throughput sequencing technology and minigene test.METHODS First,the medical history of the proband and family members of three people from two generations was collected,and the family diagram was drawn,and the audiology evaluation and ear imaging examination were performed.Genomic DNA was extracted from peripheral blood of the family.All exons,partial introns and promoters of 406 deafness gene coding regions were detected by high-throughput sequencing method for probands.Bioinformatics analysis was performed to screen suspected pathogenic mutations.Sanger sequencing verification was performed for family members according to variation sites,and minigene test was used for verification.RESULTS The clinical manifestation of the proband was bilateral severe sensorineural hearing loss.The high-throughput sequencing results showed that the homozygous mutation c.388-1G>A of USHIC resulted in a splicing mutation of amino acids.According to the ACMG guidelines,the mutation was initially identified as a suspected pathogenic mutation.Sanger sequencing showed that their parents were heterozygous phenotypeless carriers at this locus.Minigene test showed that the mutation of this site could lead to abnormal splicing of mRNA.CONCLUSION The new mutation of USHIC gene intron found in this family was pathogenic mutation,and the probands were deaf due to splicing mutation of USHIC gene intron.

关 键 词：USHER综合征 耳聋 内含子 突变 USH1C基因 微小基因 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]