Institute of Pediatrics, Children's Hospital of Fudan University, the Shanghai Key Laboratory of Medical Epigenetics, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, 200032, China.
Human Phenome Institute, Fudan University, Shanghai, 200438, China.
Genome Biol. 2023 Apr 6;24(1):68. doi: 10.1186/s13059-023-02894-0.
Oocyte maturation arrest and early embryonic arrest are important reproductive phenotypes resulting in female infertility and cause the recurrent failure of assisted reproductive technology (ART). However, the genetic etiologies of these female infertility-related phenotypes are poorly understood. Previous studies have mainly focused on inherited mutations based on large pedigrees or consanguineous patients. However, the role of de novo mutations (DNMs) in these phenotypes remains to be elucidated.
To decipher the role of DNMs in ART failure and female infertility with oocyte and embryo defects, we explore the landscape of DNMs in 473 infertile parent-child trios and identify a set of 481 confident DNMs distributed in 474 genes. Gene ontology analysis reveals that the identified genes with DNMs are enriched in signaling pathways associated with female reproductive processes such as meiosis, embryonic development, and reproductive structure development. We perform functional assays on the effects of DNMs in a representative gene Tubulin Alpha 4a (TUBA4A), which shows the most significant enrichment of DNMs in the infertile parent-child trios. DNMs in TUBA4A disrupt the normal assembly of the microtubule network in HeLa cells, and microinjection of DNM TUBA4A cRNAs causes abnormalities in mouse oocyte maturation or embryo development, suggesting the pathogenic role of these DNMs in TUBA4A.
Our findings suggest novel genetic insights that DNMs contribute to female infertility with oocyte and embryo defects. This study also provides potential genetic markers and facilitates the genetic diagnosis of recurrent ART failure and female infertility.
卵母细胞成熟阻滞和早期胚胎阻滞是导致女性不孕的重要生殖表型，也是辅助生殖技术（ART）反复失败的原因。然而，这些与女性不孕相关表型的遗传病因尚不清楚。以前的研究主要集中在基于大谱系或近亲患者的遗传突变上。然而，新生突变（DNMs）在这些表型中的作用仍有待阐明。
为了解析 DNMs 在卵母细胞和胚胎缺陷导致的 ART 失败和女性不孕中的作用，我们探索了 473 个不孕亲子三体型中的 DNMs 图谱，并鉴定出了一组分布在 474 个基因中的 481 个可信的 DNMs。基因本体论分析表明，具有 DNMs 的鉴定基因富集在与减数分裂、胚胎发育和生殖结构发育等女性生殖过程相关的信号通路中。我们对 Tubulin Alpha 4a（TUBA4A）中具有代表性的基因的 DNMs 功能进行了检测，该基因在不孕亲子三体型中具有最显著的 DNMs 富集。TUBA4A 中的 DNMs 破坏了 HeLa 细胞中微管网络的正常组装，并且 DNM TUBA4A cRNAs 的微注射导致小鼠卵母细胞成熟或胚胎发育异常，这表明这些 DNMs 在 TUBA4A 中具有致病作用。
我们的研究结果提供了新的遗传见解，即 DNMs 导致卵母细胞和胚胎缺陷的女性不孕。本研究还提供了潜在的遗传标志物，有助于反复发生的 ART 失败和女性不孕的遗传诊断。
