人类卵母细胞的单细胞 RNA 测序揭示了与琼脂样透明带相关的差异转录组图谱。
Single-cell RNA sequencing of human oocytes reveals a differential transcriptomic profile associated with agar-like zona pellucida.
机构信息
Reproductive Medical Center, Henan Province Key Laboratory for Reproduction and Genetics, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, People's Republic of China.
出版信息
J Ovarian Res. 2024 Jun 26;17(1):132. doi: 10.1186/s13048-024-01463-8.
BACKGROUND
Agar-like zona pellucida (ZP) is the most common type of abnormal ZP, and is one of the causes of low fertility or infertility. However, the molecular mechanism of agar-like ZP is unclear. Single-cell RNA-sequencing (scRNA-seq) analysis was used to assess the cellular and molecular landscape of oocytes with agar-like ZP.
METHODS
Human metaphase I (MI) oocytes were collected from four patients with agar-like ZP and four healthy donors. Total RNA was isolated, cDNA was synthesized, and libraries were generated and subsequently sequenced on a HiSeq 2500 instrument. The scRNA-seq data were analyzed with R software.
RESULTS
We identified 1320 genes that were differentially expressed between agar-like ZP oocytes and healthy donor oocytes. Gene Ontology term enrichment results showed that the genes downregulated in agar-like ZP oocytes were significantly related to extracellular matrix organization, while the genes upregulated in agar-like ZP oocytes were significantly related to the regulation of response to DNA damage stimulus. The Kyoto Encyclopedia of Genes and Genomes enrichment results showed that genes were enriched in the ECM-receptor interaction pathway and focal adhesion pathway. Other signaling pathways important in oocyte development were also enriched, such as PI3K-Akt. Differential expression analysis identified UBC, TLR4, RELA, ANXA5, CAV1, KPNA2, CCNA2, ACTA2, FYN and ITGB3 as genetic markers of oocytes with agar-like ZP.
CONCLUSIONS
Our findings suggest that agar-like ZP oocytes exhibit significant downregulation of genes involved in the ECM-receptor interaction signaling pathway and focal adhesion pathway, which could lead to aberrant ZP formation, while the upregulated genes were significantly related to regulation of the response to DNA damage stimulus. Agar-like ZP formation may interfere with the normal exchange of signals between oocytes and perivitelline granulosa cells, thereby preventing cumulus cells from participating in oocyte DNA damage repair and causing MI arrest.
背景
胶样透明带（ZP）是最常见的异常 ZP 类型之一，也是导致低生育力或不孕的原因之一。然而，胶样 ZP 的分子机制尚不清楚。单细胞 RNA 测序（scRNA-seq）分析用于评估具有胶样 ZP 的卵母细胞的细胞和分子特征。
方法
从四名胶样 ZP 患者和四名健康供体中收集人中期 I（MI）卵母细胞。分离总 RNA，合成 cDNA，生成文库，然后在 HiSeq 2500 仪器上进行测序。使用 R 软件分析 scRNA-seq 数据。
结果
我们鉴定出 1320 个在胶样 ZP 卵母细胞和健康供体卵母细胞之间差异表达的基因。GO 术语富集结果表明，胶样 ZP 卵母细胞中下调的基因与细胞外基质组织显著相关，而上调的基因与 DNA 损伤刺激反应的调控显著相关。KEGG 富集结果表明，基因在 ECM-受体相互作用通路和焦点黏附通路中富集。卵母细胞发育中其他重要的信号通路也被富集，如 PI3K-Akt。差异表达分析鉴定出 UBC、TLR4、RELA、ANXA5、CAV1、KPNA2、CCNA2、ACTA2、FYN 和 ITGB3 作为胶样 ZP 卵母细胞的遗传标志物。
结论
我们的研究结果表明，胶样 ZP 卵母细胞中 ECM-受体相互作用信号通路和焦点黏附通路相关基因显著下调，可能导致异常 ZP 形成，而上调的基因与 DNA 损伤刺激反应的调控显著相关。胶样 ZP 的形成可能干扰卵母细胞与周围颗粒细胞之间的信号正常交换，从而阻止卵丘细胞参与卵母细胞 DNA 损伤修复，并导致 MI 阻滞。
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