Department of Obstetrics and Gynecology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
NHC Key Laboratory of Study on Abnormal Gametes and Reproductive Tract (Anhui Medical University), Hefei, Anhui, China.
J Med Genet. 2024 May 21;61(6):553-565. doi: 10.1136/jmg-2023-109766.
The association between the variants and human infertility remains unclear, as only one homozygous missense variant of was found to be associated with oligoasthenoteratozoospermia (OAT).
Whole-exome sequencing and Sanger sequencing were employed to identify potential pathogenic variants of in infertile men. Histology, immunofluorescence, immunoblotting and ultrastructural analyses were conducted to clarify the structural and functional abnormalities of sperm in mutated patients. -knockout mice were generated using the CRISPR-Cas9 system. Total RNA-seq and single-cell RNA-seq (scRNA-seq) analyses were used to elucidate the underlying molecular mechanisms, followed by validation through quantitative RT-PCR and immunostaining. Intracytoplasmic sperm injection (ICSI) was also used to assess the efficacy of clinical treatment.
Bi-allelic variants were identified in five unrelated Chinese individuals with OAT, including homozygous loss-of-function variants in two consanguineous families. Notably, besides reduced concentrations and impaired motility, a significant occurrence of acrosomal hypoplasia was detected in multiple spermatozoa among five patients. Using the -deficient mice, we further elucidate the pivotal role of TDRD6 in spermiogenesis and acrosome identified. In addition, the mislocalisation of crucial chromatoid body components DDX4 (MVH) and UPF1 was also observed in round spermatids from patients harbouring variants. ScRNA-seq analysis of germ cells from a patient with variants revealed that TDRD6 regulates mRNA metabolism processes involved in spermatid differentiation and cytoplasmic translation.
Our findings strongly suggest that TDRD6 plays a conserved role in spermiogenesis and confirms the causal relationship between variants and human OAT. Additionally, this study highlights the unfavourable ICSI outcomes in individuals with bi-allelic variants, providing insights for potential clinical treatment strategies.
变体与人类不育之间的关联尚不清楚，因为仅发现一个 的纯合错义变体与少精子症伴生精障碍（OAT）有关。
采用外显子组测序和 Sanger 测序鉴定不育男性中 的潜在致病性变体。进行组织学、免疫荧光、免疫印迹和超微结构分析，以阐明突变患者精子的结构和功能异常。使用 CRISPR-Cas9 系统生成 -/- 敲除小鼠。进行总 RNA-seq 和单细胞 RNA-seq（scRNA-seq）分析以阐明潜在的分子机制，并通过定量 RT-PCR 和免疫染色进行验证。还进行了胞质内精子注射（ICSI）以评估临床治疗的效果。
在 5 名 OAT 无关的中国个体中鉴定出双等位基因 变体，包括 2 个近亲家庭的纯合失活变体。值得注意的是，除了浓度降低和运动能力受损外，在 5 名患者的多个精子中还发现顶体发育不全。使用 -/- 小鼠，我们进一步阐明了 TDRD6 在精子发生和顶体中的关键作用。此外，还观察到患者携带 变体的圆形精子中关键染色质体成分 DDX4（MVH）和 UPF1 的定位错误。来自患者的生殖细胞的 scRNA-seq 分析显示，TDRD6 调节涉及精子分化和细胞质翻译的 mRNA 代谢过程。
我们的研究结果强烈表明 TDRD6 在精子发生中发挥保守作用，并证实了 变体与人类 OAT 之间的因果关系。此外，该研究突出了具有双等位基因 变体的个体在 ICSI 中的不良结局，为潜在的临床治疗策略提供了见解。
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