A Targeted, Next-Generation Genetic Sequencing Study on Tetralogy of Fallot, Combined With Cleft Lip and Palate.
A Targeted, Next-Generation Genetic Sequencing Study on Tetralogy of Fallot, Combined With Cleft Lip and Palate.
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法洛四联症合并唇腭裂的靶向下一代基因测序研究
DOI：
10.1097/scs.0000000000003598
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发表时间：
2017-06
期刊：
影响因子：
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通讯作者：
Zhao T
中科院分区：
文献类型：
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作者：
Liu L;Bu H;Yang Y;Tan Z;Zhang F;Hu S;Zhao T
背景：先天性心脏病（CHD）和唇腭裂（CLP）是目前婴幼儿最常见的结构畸形。许多基因已被调查参与CHD与CLP。下一代靶向测序可以快速分析大量遗传信息，从而解决这个问题。方法：作者为先前与CHD或CLP有关的455个基因设计了一个靶向的下一代测序基因组。单一受试者患者作为遗传源。影响蛋白质编码区的变异被归类到计算机中，并通过数据库进行筛选，如单核苷酸多态性数据库，炎黄，外显子组测序计划和1000个基因组计划。然后，作者通过PolyPhen-2，SIFT和Mutation Taster预测了基因突变的功能。为确定相关的疾病基因，作者查阅了PubMed上的相关文献。最后，通过桑格测序验证该变体。结果如下：通过下一代靶向测序，在法洛四联症合并CLP患者中成功发现了总共1520个突变。其中ZNF 528、PVRL 2、亚甲基四氢叶酸还原酶[MTHFR]、EVC 2、DAND5、CCDC 39等6个基因突变在单核苷酸多态性数据库、炎黄工程、外显子组测序计划和千基因组计划中均未发现。通过SIFT、PolyPhen-2和Mutation Taster，四个基因（ZNF 528、PVRL 2、EVC 2、CCDC 39）均被预测为"耐受的"、"良性的"或"多态的"。DAND5基因分别被PolyPhen-2和Mutation Taster预测为“可能具有破坏性”和“致病性”，但SIFT程序预测这种突变是“可耐受的”。同样，MTHFR基因突变分别被SIFT、PolyPhen-2和Mutation Taster预测为"破坏性"、"可能破坏性"和"致病性"。PubMed上未见MTHFR基因突变（c.G586A，p.G196S）的相关报道。总结：使用靶向的下一代测序技术，作者首次确定了MTHFR基因中的突变（c.G586A，p.G196S）可能是患者TOF和CLP的原因。
Background: Congenital heart disease (CHD), plus cleft lip and palate (CLP) are currently the most common types of structural malformation in infants. Many genes have been investigated for their involvement in CHD with CLP. Targeted next-generation sequencing can analyze large amounts of genetic information rapidly, and thus address this question. Methods: The authors designed a targeted, next-generation sequencing gene panel for 455 genes previously implicated in CHD or CLP. A single-subject patient served as a genetic source. Variants that affect protein-coding regions were classified into silico and filtered through databases, such as the Single-Nucleotide Polymorphism Database, Yan Huang, the Exome Sequencing Project, and the 1000 Genomes Project. The authors then predicted the function of gene mutations by PolyPhen-2, SIFT, and Mutation Taster. To confirm the related disease genes, the authors surveyed relevant literature on PubMed. Finally, the variant was verified by Sanger sequencing. Results: A total of 1520 mutations were successfully found in a patient using combined tetralogy of Fallot and CLP by the targeted next-generation sequencing. However, there were 6 gene mutations (ZNF528, PVRL2, methylenetetrahydrofolate reductase [MTHFR], EVC2, DAND5, CCDC39) that were not found on Single-Nucleotide Polymorphism Database, Yan Huang, Exome Sequencing Project, and 1000 Genomes Project. Four genes (ZNF528, PVRL2, EVC2, CCDC39) were all predicted to be “tolerated,” “benign,” or “polymorphic” by SIFT, PolyPhen-2, and Mutation Taster. The DAND5 gene was predicted to be “possibly damaging” and “disease causing” respectively by PolyPhen-2 and Mutation Taster, but the SIFT program predicted this mutation to be “tolerated.” Likewise, the MTHFR gene mutation was predicted to be “damaging,” “possibly damaging,” and “disease causing” respectively by SIFT, PolyPhen-2, and Mutation Taster. There is no relevant report about MTHFR gene mutation (c.G586A, p.G196S) on PubMed. Conclusion: Using targeted, next-generation sequencing technology, the authors identified for the first time a mutation (c.G586A, p.G196S) in the MTHFR gene as a possible cause of TOF and CLP in a patient.