全外显子家系测序在WDR35基因复合杂合变异导致SRPS-5胎儿产前诊断中的价值

doi:10.3969/j.issn.1673-8640.2022.010.004

检验医学 ›› 2022, Vol. 37 ›› Issue (10): 928-933.DOI: 10.3969/j.issn.1673-8640.2022.010.004

全外显子家系测序在WDR35基因复合杂合变异导致SRPS-5胎儿产前诊断中的价值

赵旭亮¹, 田瑞霞², 施友文², 俞敏², 焦鎏鎏³, 朱复希¹()

1.安徽医科大学第一附属医院妇产科生殖医学中心，安徽合肥 230022
2.中国人民解放军联勤保障部队第901医院妇产科，安徽合肥 230031
3.中国人民解放军联勤保障部队第901医院影像科，安徽合肥 230031

收稿日期:2021-05-01 修回日期:2021-08-11 出版日期:2022-10-30 发布日期:2022-11-14
通讯作者: 朱复希
作者简介:朱复希，E-mail：fxzhu@163.com。
赵旭亮，男，1987年生，学士，主管技师，主要从事遗传缺陷及分子诊断研究。
基金资助:
安徽省自然科学基金资助项目(1908085J28)

Role of the prenatal diagnosis of SRPS-5 neonate caused by compound heterozygous mutation of WDR35 by Trio-WES

ZHAO Xuliang¹, TIAN Ruixia², SHI Youwen², YU Min², JIAO Liuliu³, ZHU Fuxi¹()

1. Reproductive Medical Center，Department of Obstetrics and Gynecology，the First Affiliated Hospital of Anhui Medical University，Hefei 230022，Anhui，China
2. Department of Obstetrics and Gynecology，the No. 901 Hospital of the Joint Service of the People's Liberation Army，Hefei 230031，Anhui，China
3. Department of Radiology，the No. 901 Hospital of the Joint Service of the People's Liberation Army，Hefei 230031，Anhui，China

Received:2021-05-01 Revised:2021-08-11 Online:2022-10-30 Published:2022-11-14
Contact: ZHU Fuxi

摘要/Abstract

摘要：

目的探讨全外显子家系测序（Trio-WES）在诊断WDR35基因复合杂合变异导致短肋-多指综合征5型（SRPS-5）中的应用价值。方法收集1例引产儿产前诊断的临床资料，采用Trio-WES对引产儿皮肤组织及其父母外周血进行基因测序，采用SIFT、PolyPhen-2及Mutation Taster软件对筛选出的可疑变异进行生物危害性分析，并建立模型预测变异可能导致的蛋白结构变化。结果引产儿母亲孕22⁺³周时的超声检查结果提示胎儿四肢长骨发育水平明显滞后，伴双侧肱骨、股骨弯曲，胸围小且胸腔狭窄，肋骨短且双肺受压变小。基因测序提示胎儿WDR35基因存在复合杂合变异，分别为父亲携带的c.799G>A/p.Val267Met杂合变异与母亲携带的chr2：20151176-20151245杂合缺失变异。检索Pubmed和HGMD数据库均未发现该变异相关报道，属WDR35基因的新变异，其中变异位点c.799G>A/p.Val267Met在多物种间高度保守，蛋白结构预测提示可能改变局部空间稳定性。结论 WDR35基因缺陷可能是患儿长骨明显短且弯曲，短肋伴肺发育不良等症状的致病因素。Trio-WES可精确诊断胎儿期SRPS-5。

关键词: WDR35基因, 短肋-多指综合征, 短肋-多指综合征5型, 全外显子家系测序, 产前诊断

Abstract:

Objective To investigate the role of prenatal diagnosis of a neonate of short-rib polydactyly syndrome-5（SRPS-5） induced by a compound heterozygous mutation of WDR35 by trio-whole exome sequencing （trio-WES）. Methods The clinical data of prenatal diagnosis of a neonate were collected. The genes of induced labor tissues and parents were determined by trio-WES. The biohazard analysis of the suspicious variant was carried out by using SIFT，PolyPhen-2 and Mutation Taster software，and the model was established to predict the protein structure changes caused by the mutation. Results At 22⁺³ weeks of gestation，ultrasound examination showed that fetal limb long bone had developmental delay，with bilateral humerus and femur bending，narrow chest cavity and reduced chest circumference，shortened ribs and compressed lungs. The gene test showed that there were compound heterozygous mutations in WDR35 in fetus，which were heterozygous mutation in father（c.799G>A/p.Val267Met） and loss of heterozygous mutation in chr2：20151176-2015124 in mother. The 2 mutations searched from PubMed and HGMD databases had not been reported. There were new mutations of WDR35. The c.799G>A/p.Val267Met was highly conserved among species，and the prediction of protein structure suggested that it might change the local spatial stability. Conclusions The defect of WDR35 may be the pathogenic factor of the fetal long bone shortening with bending，short rib with lung dysplasia and other symptoms，and the accurate diagnosis of SRPS-5 can be made by trio-WES.

Key words: WDR35, Short rib-polydactyly syndrome, Short-rib thoracic dysplasia 5, Trio-whole exome sequencing, Prenatal diagnosis

赵旭亮, 田瑞霞, 施友文, 俞敏, 焦鎏鎏, 朱复希. 全外显子家系测序在WDR35基因复合杂合变异导致SRPS-5胎儿产前诊断中的价值[J]. 检验医学, 2022, 37(10): 928-933.

ZHAO Xuliang, TIAN Ruixia, SHI Youwen, YU Min, JIAO Liuliu, ZHU Fuxi. Role of the prenatal diagnosis of SRPS-5 neonate caused by compound heterozygous mutation of WDR35 by Trio-WES[J]. Laboratory Medicine, 2022, 37(10): 928-933.

图/表 5

图1 本例胎儿引产前、后的影像学检查结果注：（a）产前超声检查示四肢管状骨明显较短、弯曲，尤以股骨、肱骨明显，呈“电话听筒”状，干骺端粗大，不规则扩张，呈“喇叭口”状；（b）产前超声检查示胸腔狭窄，胸围小，心胸比值大，未达到孕中期>60%的诊断标准；（c）产前超声检查示胎儿皮肤增厚，水肿，浆膜腔积液，宫内姿势和运动异常，羊水过多；（d）引产儿CT三维重建示肢骨、中肱骨、尺桡骨、股骨、胫腓骨短小并有弯曲，以肱骨、股骨弯曲明显，肋骨短且胸腔狭窄，椎骨大小正常，但椎体稍侧弯。

图2 引产儿家系图

图3 先证者（引产儿）及其父母WDR35基因变异分析注：（a）c.799G>A/p.Val267Met变异测序图；（b）chr2：20151176-20151245杂合缺失的RT-qPCR结果。

图4 不同物种之间WDR35蛋白的同源性分析注：WDR35蛋白靠近N'-端有7个β-螺旋桨状结构紧密相连组成WD40样重复结构域，c.799G>A/p.Val267Met变异发生在WD40结构域，Val267在不同物种间高度保守；chr2：20151176-20151245杂合缺失发生在WD40样重复结构域，LOF变异可能导致蛋白表达量降低。

图5 蛋白结构预测分析结果注：（a）野生型；（b）突变型；突变c.799G>A/p.Val267Met与Val286、Gln287增加了额外氢键（绿色虚线），可能导致空间结构改变。

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全外显子家系测序在WDR35基因复合杂合变异导致SRPS-5胎儿产前诊断中的价值

Role of the prenatal diagnosis of SRPS-5 neonate caused by compound heterozygous mutation of WDR35 by Trio-WES

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