Precone Vincenza, Notarangelo Angelantonio, Marceddu Giuseppe, D'Agruma Leonardo, Cannarella Rossella, Calogero Aldo E, Cristofoli Francesca, Guerri Giulia, Paolacci Stefano, Castori Marco, Bertelli Matteo
MAGI Euregio, Bolzano, Italy.
Division of Medical Genetics, IRCCS Casa Sollievo della Sofferenza Foundation, San Giovanni Rotondo, Foggia, Italy.
Minerva Endocrinol (Torino). 2022 Mar;47(1):4-10. doi: 10.23736/S2724-6507.21.03477-1. Epub 2021 May 14.
Infertility is a disorder of the male and/or female reproductive system, characterized by failure to establish a clinical pregnancy after 12 months of regular unprotected sexual intercourse. On a world basis, about one in six couples are affected by infertility during their reproductive lifespan. Despite a comprehensive diagnostic work-up, infertility in about 50% of couples remains idiopathic. In this context, a next-generation sequencing (NGS) approach has been suggested to increase diagnostic yield. Accordingly, this study aimed to evaluate the effectiveness of a custom-made NGS gene panel for the simultaneous genetic diagnosis of both partners of a large population of infertile couples.
We developed a custom-made NGS panel for 229 genes associated with male and female infertility. The panel targeted exons and their flanking regions and was used to screen 99 couples with idiopathic infertility.
NGS sequencing revealed five pathogenic variants in six couples and 17 likely pathogenic variants or variants with uncertain significance (VUS). The pathogenic variants were identified in the following genes: GNRHR, CCDC39, DNAH5, and CCDC103; likely pathogenic variants were identified in TAC3, PROKR2, and CFTR; VUS were identified in CATSPER2, FGFR1, LRRC6, DNAH5, DNAH11, TGFBR3, and DNAI1.
The panel of genes designed for this study allowed the identification of pathogenic gene mutations and the presence of VUS in 6.1% and 17.2%, respectively, of couples with idiopathic infertility. This is the first study to successfully apply an NGS-based genetic screening including 229 genes known to play a role in both male and female infertility.
不孕症是男性和/或女性生殖系统的一种疾病，其特征是在规律无保护性交12个月后未能实现临床妊娠。在全球范围内，约六分之一的夫妇在其生殖寿命期间受到不孕症的影响。尽管进行了全面的诊断检查，但约50%的夫妇的不孕症仍为特发性。在此背景下，有人提出采用下一代测序（NGS）方法来提高诊断率。因此，本研究旨在评估定制的NGS基因panel对大量不孕夫妇双方进行同时基因诊断的有效性。
我们开发了一个针对与男性和女性不孕症相关的229个基因的定制NGS panel。该panel靶向外显子及其侧翼区域，并用于筛查99对特发性不孕夫妇。
NGS测序在6对夫妇中发现了5个致病变异以及17个可能的致病变异或意义未明的变异（VUS）。在以下基因中鉴定出了致病变异：促性腺激素释放激素受体（GNRHR）、卷曲螺旋结构域蛋白39（CCDC39）、动力蛋白5（DNAH5）和卷曲螺旋结构域蛋白103（CCDC103）；在促甲状腺激素释放激素3（TAC3）、促动力蛋白受体2（PROKR2）和囊性纤维化跨膜传导调节因子（CFTR）中鉴定出了可能的致病变异；在阳离子通道精子相关蛋白2（CATSPER2）、成纤维细胞生长因子受体1（FGFR1）、富含亮氨酸重复序列蛋白6（LRRC6）、动力蛋白5（DNAH5）、动力蛋白11（DNAH11）、转化生长因子β受体3（TGFBR3）和动力蛋白轴抑制中间链1（DNAI1）中鉴定出了VUS。
本研究设计的基因panel在分别6.1%和17.2%的特发性不孕夫妇中鉴定出了致病基因突变和VUS的存在。这是第一项成功应用基于NGS的基因筛查研究，该筛查涵盖了已知在男性和女性不孕症中起作用的229个基因。
