Marić Tihana, Castillo-Madeen Helen, Klarić Monika Logara, Barišić Antun, Trgovec-Greif Lovro, Murphy Mark W, Juchnewitsch Anna-Grete, Lillepea Kristiina, Dutta Avirup, Žunić Lucija, Stendahl Alexandra M, Punab Margus, Pomm Kristjan, Mendoza Daniel M, Lopes Alexandra M, Šorgić Ana Merkler, Vugrek Oliver, Gonçalves Joao, Almstrup Kristian, Aston Kenneth I, Belužić Robert, Ježek Davor, Bertoša Branimir, Laan Maris, Bojanac Ana Katušić, Conrad Donald F, Barbalić Maja
Scientific Centre of Excellence for Reproductive and Regenerative Medicine, University of Zagreb School of Medicine, Šalata 2, Zagreb 10000, Croatia.
Department of Medical Biology, University of Zagreb School of Medicine, Šalata 2, Zagreb 10000, Croatia.
Hum Mol Genet. 2025 Mar 7;34(6):481-491. doi: 10.1093/hmg/ddae197.
The most severe form of male infertility is idiopathic non-obstructive azoospermia (NOA), a complete sperm absence in the ejaculate. We performed exome sequencing in the Croatian infertile brothers with NOA and found a variant in DMRT1 (Doublesex and mab-3 related transcription factor 1) gene that was further assessed by the EMSA assay and molecular dynamic simulations. We additionally screened for DMRT1 mutations in 1940 infertile men diagnosed with spermatogenic failure, 644 normozoospermic controls, and 105 females with primary ovarian insufficiency (POI) recruited to the GEnetics of Male INfertility Initiative (GEMINI) or Estonian Andrology (ESTAND) cohorts. DMRT1 p.Pro74Leu (chr9:g.842059C > T) variant was detected in infertile brothers in the highly conserved position within the DNA binding DM domain of the protein. EMSA assay showed reduced DNA binding of DMRT1P74L and molecular dynamic simulations showed differences in structural and dynamical properties between the wild type protein and DMRT1P74L. Plausible disease-causing DMRT1 variants were only identified in infertile men (13/1940; 0.67%), and none in 639 fertile controls. Burden testing showed an excess of rare deleterious DM domain mutations in the infertility cohort compared to gnomAD v.4.0 population-based controls (Fisher's exact test, p = 1.44 x 10-5). Three rare deleterious variants in DMRT1 were found in 104 cases of POI. The findings of this study strengthen the evidence of DMRT1 variants being a causal factor for male infertility and provide the distribution of likely pathogenic variants across the gene. This is also the first study to suggest that DMRT1 variants may also be linked to POI.
男性不育最严重的形式是特发性非梗阻性无精子症（NOA），即射精中完全没有精子。我们对克罗地亚患有无精子症的不育兄弟进行了外显子组测序，在双性和 mab-3 相关转录因子 1（DMRT1）基因中发现了一个变异，该变异通过电泳迁移率变动分析（EMSA）试验和分子动力学模拟进行了进一步评估。我们还在参与男性不育遗传学倡议（GEMINI）或爱沙尼亚男科学（ESTAND）队列的 1940 名被诊断为生精功能衰竭的不育男性、644 名正常精子症对照以及 105 名原发性卵巢功能不全（POI）女性中筛查了 DMRT1 突变。在不育兄弟中检测到 DMRT1 p.Pro74Leu（chr9:g.842059C>T）变异，该变异位于该蛋白质 DNA 结合 DM 结构域内的高度保守位置。EMSA 试验显示 DMRT1P74L 与 DNA 的结合减少，分子动力学模拟显示野生型蛋白质与 DMRT1P74L 在结构和动力学特性上存在差异。仅在不育男性中鉴定出可能致病的 DMRT1 变异（13/1940；0.67%），在 639 名生育对照中未发现。与基于 gnomAD v.4.0 人群的对照相比，负荷测试显示不育队列中罕见有害 DM 结构域突变过多（Fisher 精确检验，p = 1.44×10-5）。在 104 例 POI 病例中发现了 DMRT1 的三个罕见有害变异。本研究结果加强了 DMRT1 变异是男性不育致病因素的证据，并提供了整个基因中可能致病变异的分布情况。这也是第一项表明 DMRT1 变异可能也与 POI 有关的研究。
