Differential expression of tsRNAs and miRNAs in embryo culture medium: potential impact on embryo implantation.

Purpose Can small RNA derived from embryos in conditioned embryo culture medium (ECM) influence embryo implantation?

We employed small RNA sequencing to investigate the expression profiles of transfer RNA-derived small RNA (tsRNA) and microRNA (miRNA) in ECM from high-quality and low-quality embryos. Quantitative real-time PCR was employed to validate the findings of small RNA sequencing. Additionally, we conducted bioinformatics analysis to predict the potential functions of these small RNAs in embryo implantation. To establish the role of tiRNA-1:35-Leu-TAG-2 in embryonic trophoblast cell adhesion, we utilized co-culture systems involving JAR and Ishikawa cells.

Our analysis revealed upregulation of nine tsRNAs and four miRNAs in ECM derived from high-quality embryos, whereas 37 tsRNAs and 12 miRNAs exhibited upregulation in ECM from low-quality embryos. The bioinformatics analysis of tsRNA, miRNA, and mRNA pathways indicated that their respective target genes may play pivotal roles in both embryo development and endometrial receptivity. Utilizing tiRNA mimics, we demonstrated that the prominently expressed tiRNA-1:35-Leu-TAG-2 in the low-quality ECM group can be internalized by Ishikawa cells. Notably, transfection of tiRNA-1:35-Leu-TAG-2 into Ishikawa cells reduced the attachment rate of JAR spheroids.

Our investigation uncovers significant variation in the expression profiles of tsRNAs and miRNAs between ECM derived from high- and low-quality embryos. Intriguingly, the release of tiRNA-1:35-Leu-TAG-2 by low-quality embryos detrimentally affects embryo implantation and endometrial receptivity. These findings provide fresh insights into understanding the molecular foundations of embryo-endometrial communication. Similar content being viewed by others Data availability Additional data are available from the corresponding author on reasonable request.

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We thank Figdraw (www.figdraw.com) for expert assistance in the pattern drawing. Funding This work is supported by the Scientific Research Project of Hubei Provincial Health Commission (No. WJ2021M162), the Innovation and Cultivation Fund of Zhongnan Hospital (No. CXPY2022010), and the Basic and Clinical Medical Research Joint Fund of Zhongnan Hospital, Wuhan University (No. ZNLH202206). Author information Authors and Affiliations Contributions Designed the study and edited the final text: Yao Xiong, Ling Ma. Performed the experiments: Lei Shi, Zihan Wang, Zhidan Hong, Yanhong Mao. Wrote the manuscript and prepared the figures: Ling Ma, Ming Zhang. Contributed to manuscript revision and critical discussion: Ma Ling, Chun Zhou, Yao Xiong. Corresponding author Ethics declarations Consent for publication All authors consent for publication. Competing interests The authors declare no competing interests. Additional information Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Yao Xiong and Lei Shi were the co-first authors. Supplementary Information Below is the link to the electronic supplementary material. 10815_2024_3034_MOESM1_ESM.tif (download TIF ) Supplementary file1 RT-qPCR validation of miRNAs. qRT-PCR validation of miR-302 and miR-223 family members’ expression in high- and low-quality embryo culture medium compared to control medium. *P<0.05, **P<0.01. (TIF 5111 KB) Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Xiong, Y., Shi, L., Zhang, M. et al. Differential expression of tsRNAs and miRNAs in embryo culture medium: potential impact on embryo implantation. J Assist Reprod Genet 41, 781–793 (2024). https://doi.org/10.1007/s10815-024-03034-8 Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s10815-024-03034-8