MicroRNAs secreted by human embryos could be potential biomarkers for clinical outcomes of assisted reproductive technology.
MicroRNAs secreted by human embryos could be potential biomarkers for clinical outcomes of assisted reproductive technology.
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人类胚胎分泌的 MicroRNA 可能成为辅助生殖技术临床结果的潜在生物标志物
DOI：
10.1016/j.jare.2021.01.003
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发表时间：
2021-07
影响因子：
10.7
通讯作者：
Xiang W
中科院分区：
文献类型：
--
作者：
Fang F;Li Z;Yu J;Long Y;Zhao Q;Ding X;Wu L;Shao S;Zhang L;Xiang W
MicroRNAs(MiRNAs)是包括胚胎植入和发育在内的许多生物学功能的重要调节因子。最近，有报道称，生物液中的miRNAs对生理和病理过程具有预测作用。在本研究中，我们旨在研究人类胚胎在培养液中分泌的miRNAs是否可以作为胚胎生物标志物。培养基取自生殖医学中心知情同意患者的胚胎。应用高通量miRNA测序方法检测人胚胎培养液中的miRNA图谱。在生物信息学分析和筛选差异表达的miRNAs后，随后进行定量实时聚合酶链式反应(qRT-PCR)以进一步验证混合样本的测序结果。此外，我们还进行了液滴数字聚合酶链式反应(DdPCR)，以验证单个样本水平上的目标miRNAs。对差异表达的miRNAs进行接收器工作特性(ROC)分析。与妊娠失败的胚胎相比，妊娠成功的胚胎分泌到培养液中的miRNA谱不同，推测参与了多种生物学过程。经滴状数字聚合酶链式反应(DdPCR)验证，妊娠成功的卵裂胚胎培养液中hsa-miR-26b-5p和hsa-miR-21-5p的表达显著低于妊娠失败的胚胎。此外，受试者工作特征(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，我们的发现突出了人类胚胎在培养液中分泌的miRNAs的重要预测潜力，这对辅助生殖技术中的非侵入性胚胎选择具有重要意义。
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.
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