Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan 430030, China.
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan 430022, China.
J Adv Res. 2021 Jan 11;31:25-34. doi: 10.1016/j.jare.2021.01.003. eCollection 2021 Jul.
MicroRNAs (miRNAs) are important regulators of many biological functions, including embryo implantation and development. Recently, it has been reported that miRNAs in biofluids are predictive for physiological and pathological processes.
In this study, we aim to investigate whether the miRNAs secreted by human embryos in culture medium can be used as embryonic biomarkers.
The culture media were prospectively collected from embryos of patients at reproductive medicine center with informed consent. A high-throughput miRNA sequencing method was applied to detect the miRNA profiles in the human embryo culture media. After bioinformatics analysis and screening of differentially expressed miRNAs, quantitative real-time polymerase chain reaction (qRT-PCR) assay was subsequently performed to further confirm the sequencing results with mixed samples. Furthermore, we performed droplet digital PCR (ddPCR) to verify the target miRNAs at single sample level. Receiver operating characteristic (ROC) analyses were performed for differentially expressed miRNAs.
Compared with embryos with failed pregnancy, the embryos with successful pregnancy secreted different miRNA profiles into the culture media, which were predicted to be involved in multiple biological processes. Validated by droplet digital polymerase chain reaction (ddPCR), the expression of hsa-miR-26b-5p and hsa-miR-21-5p in the culture media of cleavage embryos with successful pregnancy was significantly lower than that of embryos with failed pregnancy. Moreover, the Receiver Operating Characteristic (ROC) curve analysis indicated that hsa-miR-26b-5p and hsa-miR-21-5p could serve as potential biomarkers for reproductive outcomes.
Together, our findings highlight the important predictive potential of miRNAs secreted by human embryos in culture media, which is meaningful for non-invasive embryo selection in assisted reproductive technology.
微小 RNA（miRNAs）是许多生物学功能的重要调节剂，包括胚胎着床和发育。最近有报道称，生物体液中的 miRNAs 可预测生理和病理过程。
本研究旨在探讨胚胎在培养介质中分泌的 miRNAs 是否可用作胚胎生物标志物。
本前瞻性研究收集了生殖医学中心患者胚胎的培养上清液，并获得知情同意。采用高通量 miRNA 测序方法检测人胚胎培养上清液中的 miRNA 谱。经过生物信息学分析和差异表达 miRNA 的筛选，随后用混合样本进行定量实时聚合酶链反应（qRT-PCR）检测以进一步验证测序结果。此外，我们还进行了液滴数字 PCR（ddPCR）以在单个样本水平验证靶 miRNAs。对差异表达的 miRNAs 进行了接收器工作特征（ROC）分析。
与妊娠失败的胚胎相比，妊娠成功的胚胎向培养上清液中分泌了不同的 miRNA 谱，这些 miRNA 被预测参与了多种生物学过程。经液滴数字 PCR（ddPCR）验证，成功妊娠卵裂胚胎培养上清液中 hsa-miR-26b-5p 和 hsa-miR-21-5p 的表达明显低于妊娠失败的胚胎。此外，ROC 曲线分析表明 hsa-miR-26b-5p 和 hsa-miR-21-5p 可作为生殖结局的潜在生物标志物。
总之，我们的研究结果强调了胚胎在培养介质中分泌的 miRNAs 具有重要的预测潜力，这对于辅助生殖技术中的非侵入性胚胎选择具有重要意义。
Zotero 插件
