Section of Endocrinology, Department of Medical Sciences, University of Ferrara, Via Savonarola 9, 44100, Ferrara, Italy,
Fam Cancer. 2014 Jun;13(2):273-80. doi: 10.1007/s10689-014-9702-y.
Multiple endocrine neoplasia type 1 (MEN1) syndrome is an autosomal dominant disease, characterized by parathyroid adenomas, endocrine gastroenteropancreatic tumors and pituitary adenomas, due to inactivating mutations of the MEN1 gene (chromosome 11q13). MEN1 mutations are mainly represented by nonsense, deletions/insertions, splice site or missense mutations that can be detected by direct sequencing of genomic DNA. However, MEN1 patients with large heterozygous deletions may escape classical genetic screening and may be misidentified as phenocopies, thereby hindering proper clinical surveillance. We employed a real-time polymerase chain reaction application, the TaqMan copy number variation assay, to evaluate a family in which we failed to identify an MEN1 mutation by direct sequencing, despite a clear clinical diagnosis of MEN1 syndrome. Using the TaqMan copy number variation assay we identified a large deletion of the MEN1 gene involving exons 1 and 2, in three affected family members, but not in the other nine family members that were to date clinically unaffected. The same genetic alteration was not found in a group of ten unaffected subjects, without family history of endocrine tumors. The MEN1 deletion was further confirmed by multiplex ligation-dependent probe amplification, which showed the deletion extended from exon 1 to exon 3. This new approach allowed us to correctly genetically diagnose three clinical MEN1 patients that were previously considered as MEN1 phenocopies. More importantly, we excluded the presence of genetic alterations in the unaffected family members. These results underline the importance of using a variety of available biotechnology approaches when pursuing a genetic diagnosis in a clinically suggestive setting of inherited endocrine cancer.
多发性内分泌腺瘤 1 型(MEN1)综合征是一种常染色体显性疾病,其特征为甲状旁腺瘤、内分泌胃肠胰肿瘤和垂体腺瘤,由 MEN1 基因(染色体 11q13)的失活突变引起。MEN1 突变主要表现为无义、缺失/插入、剪接位点或错义突变,可通过基因组 DNA 的直接测序检测到。然而,MEN1 患者的大片段杂合性缺失可能逃避经典的遗传筛查,可能被误诊为表型模拟,从而阻碍了适当的临床监测。我们采用了实时聚合酶链反应应用,即 TaqMan 拷贝数变异分析,来评估一个家族,尽管我们通过直接测序未能识别 MEN1 突变,但该家族有明确的 MEN1 综合征临床诊断。使用 TaqMan 拷贝数变异分析,我们在三个受影响的家族成员中鉴定出 MEN1 基因涉及外显子 1 和 2 的大片段缺失,但在其他 9 个迄今未受临床影响的家族成员中未发现。在一组 10 个无内分泌肿瘤家族史的无影响受试者中也未发现相同的遗传改变。MEN1 缺失进一步通过多重连接依赖性探针扩增证实,该缺失从外显子 1 延伸至外显子 3。这种新方法使我们能够正确地对先前被认为是 MEN1 表型模拟的三名临床 MEN1 患者进行基因诊断。更重要的是,我们排除了未受影响的家族成员存在遗传改变的可能性。这些结果强调了在具有遗传性内分泌癌临床提示的情况下,使用各种可用的生物技术方法进行遗传诊断的重要性。