新发突变单基因病家系产前诊断结果及复发性新发突变患儿的亲本来源分析  

Prenatal diagnosis of families with de novo monogenic diseases and origin analysis of recurrent de novo mutations

作　　者：谢瑶君 陈松长 张丽 刘聪 傅元政 徐晨明 Xie Yaojun;Chen Songchang;Zhang Li;Liu Cong;Fu Yuanzheng;Xu Chenming(Genetics Center of Obstetrics and Gynecology,Obstetrics&Gynecology Hospital of Fudan University,Shanghai 200011,China)

机构地区：[1]复旦大学附属妇产科医院妇产科遗传中心,上海200011

出　　处：《中华医学杂志》2025年第6期452-458,共7页National Medical Journal of China

基　　金：国家重点研发计划(2023YFC2705600,2023YFC2705601,2023YFC2705603);上海申康医院发展中心“市级医院新兴前沿技术联合攻关项目”(SHDC12023120)。

摘　　要：目的分析新发突变(DNMs)单基因病家系产前诊断结果及复发性DNMs家庭患儿的突变亲本来源。方法本研究为横断面研究。纳入2021年1月至2023年12月于复旦大学附属妇产科医院行生殖遗传咨询和产前诊断的DNMs患儿生育史家系41个。收集产前诊断结果及其他临床资料,采集患儿父母外周血、先证者样本(外周血或引产胎儿组织)以及羊水或绒毛样本。对复发性DNMs患儿生育史家系,额外收集患儿父母双方唾液及精液。行靶向高通量测序,评估患儿父母体细胞及男方生殖腺嵌合情况。对于精液中未检出致病变异的家系,利用Sanger测序寻找变异上下游单核苷酸多态性(SNP)位点并结合TA克隆技术明确变异亲本来源。结果共纳入41个家系,41例男性年龄(34.1±3.9)岁,41例女性年龄(33.0±3.9)岁,涉及致病基因32个,神经发育障碍类疾病占53.7%(22/41),遗传性肌病占12.2%(5/41),遗传性骨病占7.3%(3/41),遗传性眼病占4.9%(2/41),心血管遗传性疾病占2.4%(1/41),其他多系统疾病占19.5%(8/41)。在37个再次妊娠行产前诊断家系中,1个检出DNMs。对4个有复发性DNMs患儿生育史的家系行DNMs亲本来源分析,其中2个家系在父源精液中检出低比例嵌合,变异等位基因分数(VAF)分别为3.7%和12.8%,2个家系通过Sanger测序结合TA克隆提示为母源变异。结论DNMs具有一定复发风险。利用“靶向高通量测序+Sanger测序+TA克隆验证”的分析流程有助于寻找变异亲本来源。ObjectiveTo analyze the prenatal diagnostic results in families with de novo monogenic diseases and the mutation origins in affected children from families with reproductive history of children with recurrent de novo mutations(DNMs).MethodsThis study was a cross-sectional study.A total of 41 cases with adverse pregnancy history of de novo monogenic diseases who underwent genetic counseling and prenatal diagnosis from January 2021 to December 2023 at the Obstetrics and Gynecology Hospital of Fudan University were included.Prenatal diagnosis and other clinical data were reviewed,and peripheral blood of the parents,peripheral blood or tissue of the probands,and amniotic fluid or chorionic villus were collected.For families with reproductive history of children with recurrent DNMs,additional saliva and semen were collected from all the parents.Targeted high-throughput sequencing was performed to assess parental somatic mosaicism and male germline mosaicism.As for the cases in which the mutation was undetected in the semen,Sanger sequencing was utilized to search for single nucleotide polymorphism(SNP)sites upstream and downstream of the mutation site and clarify the mutation origins in combination with TA cloning.ResultsA total of 41 families were included,with male age of(34.1±3.9)years(41 cases)and female age of(33.0±3.9)years(41 cases).Moreover,32 causative genes were involved,with neurodevelopmental disorders,hereditary myopathies,hereditary bone diseases,hereditary ophthalmopathies,hereditary cardiovascular diseases and other multisystem diseases accounting for 53.7%(22/41),12.2%(5/41),7.3%(3/41),4.9%(2/41),2.4%(1/41),and 19.5%(8/41),respectively.One DNMs was detected in 37 families who underwent prenatal diagnosis during the second trimester.Four families with reproductive history of children with recurrent DNMs were analyzed for the mutation origins,of which two families had a low proportion of mosaicism detected in paternal semen,with variant allele fraction(VAF)of 3.7%and 12.8%,respectively,and the origins

关 键 词：产前诊断 新发突变 单基因病 靶向高通量测序 生殖腺嵌合 TA克隆 

分 类 号：R725[医药卫生—儿科]