Department of Cardiovascular Surgery, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, P.R. China.
Department of Stomatology, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, P.R. China.
Mol Med Rep. 2019 May;19(5):3831-3840. doi: 10.3892/mmr.2019.10043. Epub 2019 Mar 15.
Congenital heart disease (CHD), and cleft lip and palate (CLP) are currently the most common types of structural malformation in infants. Various methods have been used to identify the disease‑associated genes. However, targeted next‑generation sequencing (NGS) is not yet considered an option for routine use. Thus, the present study aimed to assess the safety and feasibility of using targeted NGS in patients with CHD concomitant with CLP. Between November 2015 and May 2017, a total of 17 patients with CHD concomitant with CLP, who were excluded from a diagnosis of trisomy syndrome, were selected at The Second Xiangya Hospital of Central South University (Changsha, China). Genomic DNA was extracted from peripheral blood samples of the patients. The copy number variants (CNVs) were determined by conducting a single nucleotide polymorphism (SNP) array with Illumina HumanOmni1‑Quad Beadchip, while information on other gene mutations was obtained from targeted sequencing. The functions of gene mutations were then predicted using the PolyPhen‑2, SIFT and Mutation Taster tools. Finally, Sanger sequencing was used to verify the mutations. The results identified no pathogenic mutations in CNVs analyzed by high‑throughput SNP sequencing. Targeted NGS results demonstrated that 10 patients (58.8%) carried gene mutations, including 4 (23.5%) genetically diagnosed cases and 6 (35.3%) cases with unknown etiology. The 4 known diseases were Opitz G/BBB syndrome caused by MID1 gene mutation, Loeys‑Dietz syndrome caused by TGFBR1 gene mutation, Ritscher‑Schinzel/3C syndrome caused by KIAA0196 gene mutation and CHARGE syndrome caused by CHD7 gene mutation. The remaining 6 cases were not genetically diagnosed, although they carried candidate genes. In conclusion, the present study demonstrated that targeted NGS was an effective and accurate candidate gene detection method in patients with CHD concomitant with CLP.
先天性心脏病（CHD）和唇腭裂（CLP）是目前婴儿中最常见的结构畸形类型。已经使用了各种方法来鉴定与疾病相关的基因。然而，靶向下一代测序（NGS）尚未被认为是常规使用的选择。因此，本研究旨在评估在伴有 CLP 的 CHD 患者中使用靶向 NGS 的安全性和可行性。2015 年 11 月至 2017 年 5 月，中南大学湘雅二医院（长沙，中国）共选择了 17 例排除三体综合征诊断的伴有 CLP 的 CHD 患者。从患者外周血样本中提取基因组 DNA。通过 Illumina HumanOmni1-Quad Beadchip 进行单核苷酸多态性（SNP）阵列，确定拷贝数变异（CNVs），而其他基因突变信息则通过靶向测序获得。然后使用 PolyPhen-2、SIFT 和 Mutation Taster 工具预测基因突变的功能。最后，使用 Sanger 测序验证突变。高通量 SNP 测序分析未发现 CNV 中的致病性突变。靶向 NGS 结果表明，10 例患者（58.8%）携带基因突变，包括 4 例（23.5%）基因诊断病例和 6 例（35.3%）病因不明病例。4 种已知疾病为 MID1 基因突变引起的 Opitz G/BBB 综合征、TGFBR1 基因突变引起的 Loeys-Dietz 综合征、KIAA0196 基因突变引起的 Ritscher-Schinzel/3C 综合征和 CHD7 基因突变引起的 CHARGE 综合征。其余 6 例虽然携带候选基因，但未进行基因诊断。总之，本研究表明，靶向 NGS 是伴有 CLP 的 CHD 患者有效的、准确的候选基因检测方法。
