Li Xiaoliang, Shi Guocheng, Li Yang, Zhang Xiaoqing, Xiang Ying, Wang Teng, Li Yanxin, Chen Huiwen, Fu Qihua, Zhang Hong, Wang Bo
Pediatric Translational Medicine Institute, Shanghai Children's Medical Center Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China.
Department of Cardiothoracic Surgery, Heart Center, Shanghai Children's Medical Center Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China.
J Med Genet. 2020 May 6. doi: 10.1136/jmedgenet-2019-106608.
CNV is a vital pathogenic factor of congenital heart disease (CHD). However, few CNVs have been reported for total anomalous pulmonary venous connection (TAPVC), which is a rare form of CHD. Using case-control study, we identified 15q11.2 deletion associated with TAPVC. We then used a TAPVC trio as model to reveal possible molecular basis of 15q11.2 microdeletion.
CNVplex and Chromosomal Microarray were used to identify and validate CNVs in samples from 231 TAPVC cases and 200 healthy controls from Shanghai Children's Medical Center. In vitro cardiomyocyte differentiation of induced pluripotent stem cells from peripheral blood mononuclear cells for a TAPVC trio with paternal inherited 15q11.2 deletion was performed to characterise the effect of the deletion on cardiomyocyte differentiation and gene expression.
The 15q11.2 microdeletion was significantly enriched in patients with TAPVC compared with healthy control (13/231 in patients vs 0/200 in controls, p=5.872×10, Bonferroni adjusted) using Fisher's exact test. Induced pluripotent stem cells from the proband could not differentiate into normal cardiomyocyte. Transcriptomic analysis identified a number of differentially expressed genes in the 15q11.2 deletion carriers of the family. TAPVC disease-causing genes such as , and showed significantly higher expression in the proband compared with her healthy mother. Knockdown of TUBGCP5 could lead to abnormal cardiomyocyte differentiation.
We discovered that the 15q11.2 deletion is significantly associated with TAPVC. Gene expression profile that might arise from 15q11.2 deletion for a TAPVC family was characterised using cell experiments.
拷贝数变异（CNV）是先天性心脏病（CHD）的一个重要致病因素。然而，关于完全性肺静脉异位连接（TAPVC）（一种罕见的CHD形式）的CNV报道很少。通过病例对照研究，我们鉴定出与TAPVC相关的15q11.2缺失。然后，我们以一个TAPVC三联体为模型来揭示15q11.2微缺失可能的分子基础。
使用CNVplex和染色体微阵列技术，对上海儿童医学中心231例TAPVC病例和200例健康对照的样本进行CNV鉴定和验证。对一个父亲遗传有15q11.2缺失的TAPVC三联体的外周血单个核细胞来源的诱导多能干细胞进行体外心肌细胞分化，以表征该缺失对心肌细胞分化和基因表达的影响。
使用Fisher精确检验，与健康对照相比，15q11.2微缺失在TAPVC患者中显著富集（患者中13/231，对照中0/200，p = 5.872×10，经Bonferroni校正）。先证者的诱导多能干细胞不能分化为正常心肌细胞。转录组分析确定了该家族15q11.2缺失携带者中一些差异表达基因。与健康母亲相比，先证者中TAPVC致病基因如 、 和 表达显著更高。敲低TUBGCP5可导致心肌细胞分化异常。
我们发现15q11.2缺失与TAPVC显著相关。通过细胞实验表征了一个TAPVC家族中15q11.2缺失可能产生的基因表达谱。
