Preimplantation Genetic Diagnosis of Multiple Endocrine Neoplasia Type 2A Using Informative Markers Identified by Targeted Sequencing
Preimplantation Genetic Diagnosis of Multiple Endocrine Neoplasia Type 2A Using Informative Markers Identified by Targeted Sequencing
复制标题
使用靶向测序鉴定的信息标记对 2A 型多发性内分泌肿瘤进行植入前遗传学诊断
DOI：
10.1089/thy.2017.0200
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发表时间：
2018-03-01
期刊：
影响因子：
6.6
通讯作者：
Xu, Chenming
中科院分区：
文献类型：
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作者：
Chen, Songchang;Li, Shuyuan;Xu, Chenming
背景：修订后的甲状腺髓样癌治疗指南建议对所有育龄RET突变携带者考虑生殖选择的遗传咨询，包括植入前遗传学诊断（PGD），以避免多发性内分泌肿瘤2型（MEN2）的传播。然而，PGD的高复杂性和高成本阻碍了其广泛应用。因此，有必要建立一种简单且相对廉价的方法来促进MEN2的PGD。患者和方法：设计定制的Nimblegen EZ序列捕获阵列，捕获目标区域，包括RET基因，RET基因每侧1Mb范围。对一个MEN2A家族的三个成员（这对夫妇和父亲）进行了靶向的、基于捕获的下一代测序，以确定信息标记。胚胎的诊断是通过基于信息标记和致病突变的单倍型分析实现的。结果：根据测序结果，共检测到173个信息性标记，可供后续PGD使用。选择7个信息性标记和致病突变（RETC634Y），进行Sanger测序。通过单倍型分析，4个未遗传RET基因突变单倍型的胚胎被诊断为未受影响。一个未受影响的胚胎被移植，一个健康的婴儿在38孕周出生。结论：靶向的、基于捕获的新一代测序与Sanger测序联合用于信息标记的鉴定是一种简单有效的方法，用于单基因疾病如MEN2的PGD。
Background: The revised guidelines for the management of medullary thyroid carcinoma recommend that genetic counseling regarding reproductive options, including preimplantation genetic diagnosis (PGD), be considered for all RET mutation carriers of reproductive age to avoid the transmission of multiple endocrine neoplasia type 2 (MEN2). However, the high complexity and cost of PGD have hindered its widespread use. Thus, it is necessary to establish a simple and relatively inexpensive method to facilitate the PGD of MEN2.Patients and Methods: A customized Nimblegen EZ sequence capture array was designed to capture the targeted regions, including the RET gene, and 1Mb range on each side of the RET gene. Targeted, capture-based next-generation sequencing of three members of one family with MEN2A (the couple and the paternal father) was conducted to identify the informative markers. The diagnosis of the embryos was achieved through haplotype analysis based on informative markers and causative mutation.Results: Based on the sequencing results, 173 informative markers were detected, which were sufficient for the subsequent use for PGD. Seven informative markers and the causative mutation (RETC634Y) were selected and subjected to Sanger sequencing. Through haplotype analysis, four embryos without inheritance of the mutation haplotype of the RET gene were diagnosed as unaffected. One unaffected embryo was transferred, with one healthy baby born at 38 gestational weeks.Conclusions: Targeted, capture-based next-generation sequencing for identification of informative markers together with Sanger sequencing is an easy and efficient method for the PGD of monogenic diseases such as MEN2.