房间隔缺损家系致病基因筛查分析

姚梦瑜; 马丽晶; 孔瑞泽; 刘劼; 曹宇; 蒋立虹

doi:10.12259/j.issn.2095-610X.S20220327

房间隔缺损家系致病基因筛查分析

doi: 10.12259/j.issn.2095-610X.S20220327

姚梦瑜^{1, 2},
马丽晶³,
孔瑞泽⁴,
刘劼⁵,
曹宇^2, ,,
蒋立虹²

1.
昆明理工大学附属医院，云南昆明　650500
2.
云南省第一人民医院心脏大血管外科
3.
内分泌科
4.
普外一科
5.
再生医学研究中心，云南昆明　650032

基金项目: 国家自然科学基金资助项目（81960068）；云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目 [2018FE001（-181）]；云南省高层次卫生计生技术人才培养经费（YNWR-QNBJ-2020-263）

详细信息

作者简介:
姚梦瑜（1995～），女，江苏镇江人，在读硕士研究生，主要从事心脏大血管外科临床工作

通讯作者:
蒋立虹，E-mail：jlh15198763375@163.com；曹宇，E-mail：18687035472@163.com

中图分类号: R604
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出版历程
- 收稿日期: 2022-02-01
- 网络出版日期: 2022-02-18
- 刊出日期: 2022-03-22

Screening and Analysis of Pathogenic Genes in Families with Atrial Septal Defects

Mengyu YAO^{1, 2},
Lijing MA³,
Ruize KONG⁴,
Jie LIU⁵,
Yu CAO^{2
, ,},
Lihong JIANG²

1.
The Affiliated Hospital of Kunming University of Science and Technology，Kunming Yunnan 650500
2.
Dep.t of Cardiovascular Surgery
3.
Dept. of Endocrinology
4.
First Department of General Surgery，The First People’s Hospital of Yunnan Province， Kunming Yunnan 650032
5.
Regenerative Medicine Research Center，Kunming Yunnan 650032，China

摘要

摘要: 目的通过对一个房间隔缺损先证者的四代家系进行致病突变筛查，利用全外显子测序及Sanger测序等分析基因型和表型之间的关系。方法回顾性分析在云南省第一人民医院心脏大血管外科就诊的1例房间隔缺损患儿的临床资料，并采用全外显子测序技术对患儿及其弟弟、母亲进行遗传学检测，结合生物信息学分析，寻找导致该家系致病的遗传学病因。同时，对在该家系中发现的尾型同源盒基因4（Caudal Homebox Gene，CDX4）基因变异，进行散发ASD患者及健康人群的筛查。结果患儿心脏超声结果及术中所见符合ASD诊断。基因检测提示患儿存在CDX4基因c.C233T纯合变异，其母亲携带该基因的杂合变异，其弟弟携带该基因纯合变异。CDX4基因第1外显子第233号密码子由C突变为T（c.C233T），导致第78号氨基酸由脯氨酸变为亮氨酸（p.P78L）。在198例散发性房间隔缺损患者中，5例患者存在该基因变异，而265例健康人群中未发现该基因变异。采用Fischer精确检验分析发现CDX4基因c.C233T纯合变异与先天性心脏病相关（P = 0.014）。结论 CDX4基因c.C233T（p.P78L）纯合突变很可能是该家系的致病变异。
- 房间隔缺损 /
- 全外显子测序 /
- CDX4 /
- 基因变异
Abstract: Objective To screen the pathogenic mutations in the four generations of a proband with atrial septal defect, so as to analyze the relationship between genotype and phenotype by whole exon sequencing and Sanger sequencing. Methods Retrospectic analysis of the clinical data on a child with atrial septal defects treated in the Department of Cardiovascular Surgery, the First People’ s Hospital of Yunnan Province was conducted in tandem with genetic tests for the sick child, his younger brother and mother by means of whole-exome sequencing to find out the genetic causes of the family in combination with bioinformatics analysis. Meanwhile, sporadic ASD patients and healthy people were screened for the Caudal Homebox Gene 4 （CDX4） mutation found in the family. Results The cardiac ultrasound results of the sick child and the intraoperative findings conformed with ASD diagnosis. The genetic tests suggested that CDX4 gene c.C233T homozygous mutation occurred to the child, his mother carries heterozygous mutation of the gene and his younger brother homozygous mutation of it. Codon 233 of exon 1 of CDX4 gene mutated from C to T （c.C233T）, leading to conversion of amino acid 78 from proline to leucine （p.P78L）. Multi-species comparison showed that the locus was highly conserved. The three software programs PROVEAN, Mutation Taster and Mutation Assessor suggested that the mutation was harmful. Of the 198 patients with sporadic atrial septal defects, 5 suffered this gene mutation, but that was not found in the 265 healthy people. According to the Fischer’s Exact Test, CDX4 gene c.C233T homozygous mutation was associated with congenital heart diseases （P = 0.014）. Conclusion CDX4 gene c.C233T （p.P78L） homozygous mutation is the highly likely the pathogenic mutation of the family.
- Atrial Septal Defect /
- Whole exon sequencing /
- CDX4 /
- Gene mutation

HTML全文

图 1 ASD先证者及其家系遗传图谱

Figure 1. Pedigree of the family suffering from Atrial septal defect

下载: 全尺寸图片幻灯片

图 2 患儿及其弟弟、母亲家系Sanger测序验证结果

A：先证者IV1 c.C233T纯合变异；B：先证者弟弟IV2c.C233T纯合变异；C：先证者母亲Ⅲ2 c.C233T杂合变异。红线框出为基因编码突变区。

Figure 2. Sanger sequencing verification results of the children and their brother and mother families

下载: 全尺寸图片幻灯片

图 3 CDX4 c.C233T （p.P78L）多序列比对结果。用黑色框线框出

Figure 3. CDX4 C.233T (P.78L) multi-sequencealignment results. The amino acids at this site are outlined in black

下载: 全尺寸图片幻灯片

表 1 Sanger测序人群验证及变异的致病性分析结果

Table 1. Population validation and pathogenicity analysis results of Sanger sequencing

基因类型	房间隔缺损患者		健康人群		P
基因类型	n	百分比（%）	n	百分比（%）	P
CT	5	2.52	0	0	0.014
TT	193	97.48	265	100	0.014

下载: 导出CSV

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