Columbia University College of Physicians & Surgeons, New York, New York.
Department of Otolaryngology-Head and Neck Surgery, Columbia University Medical Center, New York, New York.
Birth Defects Res. 2017 Oct 2;109(16):1257-1267. doi: 10.1002/bdr2.1063. Epub 2017 Jul 27.
The majority of cleft lip with or without cleft palate cases appear as an isolated, nonsyndromic entity (NSCLP). With the advent of next generation sequencing, whole exome sequencing (WES) has been used to identify single nucleotide variants and insertion/deletions which cause or increase the risk of NSCLP. However, to our knowledge, there are no published studies using WES in NSCLP to investigate copy number changes (CNCs), which are a major component of human genetic variation. Our study aimed to identify CNCs associated with NSCLP in a Honduran population using WES.
WES was performed on two to four members of 27 multiplex Honduran families. CNCs were identified using two algorithms, CoNIFER and XHMM. Priority was given to CNCs that were identified in more than one patient and had variant frequencies of less than 5% in reference data sets.
WES completion was defined as >90% of the WES target at >8 × coverage and >80% of the WES target at >20 × coverage. Twenty-four CNCs that met our inclusion criteria were identified by both CoNIFER and XHMM. These CNCs were confirmed using quantitative PCR. Pedigree analysis produced three CNCs corresponding to ADH7, AHR, and CRYZ segregating with NSCLP. Two of the three CNCs implicate genes, AHR and ADH7, whose known biological functions could plausibly play a role in NSCLP.
WES can be used to detect candidate CNCs that may be involved in the pathophysiology of NSCLP. Birth Defects Research 109:1257-1267, 2017. © 2017 Wiley Periodicals, Inc.
大多数唇裂伴或不伴腭裂的病例表现为孤立的、非综合征型实体(NSCLP)。随着下一代测序技术的出现,全外显子组测序(WES)已被用于识别导致或增加 NSCLP 风险的单核苷酸变异和插入/缺失。然而,据我们所知,尚无使用 WES 研究 NSCLP 中拷贝数变化(CNC)的出版物,而 CNC 是人类遗传变异的主要组成部分。我们的研究旨在使用 WES 鉴定与洪都拉斯人群的 NSCLP 相关的 CNC。
对 27 个多重洪都拉斯家系的两到四个成员进行 WES。使用 CoNIFER 和 XHMM 两种算法识别 CNC。优先考虑在多个患者中鉴定出的 CNC,并且在参考数据集中有少于 5%的变异频率。
WES 完成定义为> 90%的 WES 目标覆盖率> 8×,> 80%的 WES 目标覆盖率> 20×。CoNIFER 和 XHMM 均鉴定出符合我们纳入标准的 24 个 CNC。这些 CNC 通过定量 PCR 得到了确认。系谱分析产生了三个 CNC,分别对应于 ADH7、AHR 和 CRYZ,与 NSCLP 分离。三个 CNC 中有两个涉及基因 AHR 和 ADH7,其已知的生物学功能可能在 NSCLP 中发挥作用。
WES 可用于检测可能参与 NSCLP 病理生理学的候选 CNC。出生缺陷研究 109:1257-1267,2017。© 2017 Wiley Periodicals, Inc.