13例先天性双侧输精管缺如不育患者的致病基因突变检测  

Detection of pathogenic gene mutations in thirteen cases of congenital bilateral absence of vas deferens infertility patients

作　　者：汤莹 张湧波 吴丹红 林炎鸿 兰风华 TANG Ying;ZHANG Yongbo;WU Danhong;LIN Yanhong;LAN Fenghua(Fujian Provincial Key Laboratory of Transplant Biology,Fuzong Clinical Medical College of Fujian Medical University(The 900th Hospital of Joint Logistic Support Force,PLA),Fuzhou 350025,China)

机构地区：[1]福建省移植生物学重点实验室,福建医科大学福总临床医学院(第九〇〇医院),福州350025

出　　处：《北京大学学报（医学版）》2024年第5期763-774,共12页Journal of Peking University:Health Sciences

基　　金：福建省自然科学基金(2020J011149、2023J011351);第九〇〇医院院立课题(2021MS03)。

摘　　要：目的:检测先天性双侧输精管缺如(congenital bilateral absence of the vas deferens,CBAVD)患者中囊性纤维化跨膜转导调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)基因和CBAVD易感基因的突变情况,探讨它们与CBAVD发病风险的相关性。方法:对13例诊断为孤立发生的CBAVD患者的致病基因CFTR及易感基因黏附型G蛋白偶联受体G2(adhesion G protein-coupled receptor G2,ADGRG2)、上皮细胞钠离子通道β亚单位(sodium channel epithelial 1 subunit beta,SCNN1B)和碳酸酐酶12(carbonic anhydrase,CA12)和溶质载体家族9成员3(solute carrier family 9 member A3,SLC9A3)行全外显子测序及Sanger测序验证,针对CFTR基因多态性位点、内含子及侧翼序列行聚合酶链式反应(polymerase chain reaction,PCR)扩增后用Sanger测序,并运用生物信息学方法对CBAVD易感基因新发突变进行保守性分析和有害性预测。对13例CBAVD患者中1例患者的家系进行遗传学分析,评估子代遗传风险。结果:外显子测序发现13例CBAVD患者中,只有6例患者检测到CFTR基因外显子突变,有6种错义突变:c.2684G>A(p.Ser895Asn)、c.4056G>C(p.Gln1352His)、c.2812G>T(p.Val938Leu)、c.3068T>G(p.Ile1023Arg)、c.374T>C(p.Ile125Thr)、c.1666A>G(p.Ile556Val),1种无义突变:c.1657C>T(p.Arg553Ter),这6例患者中有2例患者同时还存在CFTR的纯合p.V470位点,另外7例患者未检测出CFTR基因外显子区域的突变。13例CBAVD患者中,3例患者携带纯合p.V470的多态性位点,4例患者携带5T等位基因,2例患者携带TG13等位基因,10例患者携带c.-966T>G位点。4例CBAVD患者同时携带以上CFTR基因突变位点中的2~3个位点。13例患者中CBAVD易感基因突变情况:1种ADGRG2错义突变c.2312A>G(p.Asn771Ser),2种SLC9A3错义突变c.2395T>C(p.Cys799Arg)、c.493G>A(p.Val165Ile),1种SCNN1B错义突变c.1514G>A(p.Arg505His)和1种CA12错义突变c.1061C>T(p.Ala354Val),其中,SLC9A3基因的c.493G>A(p.Val165Ile)突变位点是首次在CBAVD患者中�Objective:To detect the cystic fibrosis transmembrane transduction regulator(CFTR)gene mutations and congenital bilateral absence of vas deferens(CBAVD)susceptibility gene mutations in patients with CBAVD,and to explore their association with the risk of CBAVD.Methods:Whole-exome sequencing and Sanger sequencing validation were conducted on the pathogenic genes CFTR,adhesion G protein-coupled receptor G2(ADGRG2),sodium channel epithelial 1 subunit beta(SCNN1B),carbonic anhydrase 12(CA12),and solute carrier family 9 member A3(SLC9A3)in thirteen cases of isolated CBAVD patients.The polymorphic loci,intron and flanking sequences of CFTR gene were amplified by polymerase chain reaction(PCR)followed by Sanger sequencing.Bioinformatics methods were employed for conservative analysis and deleterious prediction of novel susceptibility gene mutations in CBAVD.Genetic analysis was performed on the pedigree of one out of thirteen patients with CBAVD to evaluate the risk of inheritance in offspring.Results:Exome sequencing revealed CFTR gene exon mutations in only six of the thirteen CBAVD patients,with six missense mutations c.2684G>A(p.Ser895Asn),c.4056G>C(p.Gln1352His),c.2812G>(p.Val938Leu),c.3068T>G(p.Ile1023Arg),c.374T>C(p.Ile125Thr),c.1666A>G(p.Ile556Val)),and one nonsense mutation(c.1657C>T(p.Arg553Ter).Among these six patients,two also had the CFTR homozygous p.V470 site,additionally,mutations in CFTR gene exon regions were not detected in the remaining seven patients.Within the thirteen CBAVD patients,three carried the homozygous p.V470 polymorphic site,four carried the 5T allele,two carried the TG13 allele,and ten carried the c.-966T>G site.Four CBAVD patients simultaneously carried 2-3 of the aforementioned CFTR gene mutation sites.Susceptibility gene mutations in CBAVD among the thirteen patients included one ADGRG2 missense mutation c.2312A>G(p.Asn771Ser),two SLC9A3 missense mutations c.2395T>C(p.Cys799Arg),c.493G>A(p.Val165Ile),one SCNN1B missense mutation c.1514G>A(p.Arg505His),and one CA12 missense mutation c

关 键 词：先天性双侧输精管缺如 囊性纤维化跨膜转导调节因子 黏附型G蛋白偶联受体G2 溶质载体家族9成员3 

分 类 号：R394.1[医药卫生—医学遗传学]