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Maternal circulating exosomal miR-185-5p levels as a predictive biomarker in patients with recurrent pregnancy loss

  • Reproductive physiology and disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The aim of this study was to explore the predictive role of microRNAs (miRNAs) from maternal serum exosomes in early recurrent pregnancy loss (RPL) and the related mechanism in early pregnancy.

Methods

Maternal serum was collected from pregnant women with RPL history or women with ongoing pregnancy (OP); serum exosomes were extracted and identified. Differentially expressed (DE) miRNAs in exosomes were screened by RNA sequencing and further validated by qRT-PCR. Next, the predictive value of exosomal miRNA and the clinical indicators for subsequent miscarriage in RPL patients were evaluated. Additionally, we verified the regulatory relationship between miR-185-5p and vascular endothelial growth factor (VEGF) in decidual natural killer (dNK) cells by overloading or inhibiting the exosomal miR-185-5p level in trophoblast cells.

Results

The miRNA sequencing revealed 43 DE miRNAs between OP and RPL patients. The five most significant DE miRNAs (miR-22-3p, miR-185-5p, miR-335-3p, miR-362-5p, and miR-378a-3p) were selected for identification, and miR-185-5p was increased in RPL patients. The area under curve (AUC) of the receiver operating characteristic was 0.925 when using miR-185-5p as a biomarker for subsequent miscarriage in RPL patients. In addition, miR-185-5p in exosomes secreted from HTR-8 cells reduces VEGF expression of dNK cells.

Conclusions

The current study, for the first time, successfully constructed the correlation between maternal circulating exosomal miR-185-5p expression pattern and RPL, which may be involved in the pathogenesis of RPL by downregulating the VEGFA of dNK cells and perturbing angiogenesis at the maternal–fetal interface.

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Data availability

The data included in this article are available from the corresponding author upon reasonable request.

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Acknowledgements

We are indebted to the individuals who participated in the research. We are very grateful to nurse Rong Zhang for helping to coordinate the collection of samples and data. We thank the National Natural Science Foundation of China for supporting this project. We thank John Myers‍ at AJE for editing the language of this manuscript.

Funding

This research was supported by a grant from the National Natural Science Foundation of China (No. 81871182).

Author information

Author notes

  1. Yujing Xiong, Zheng Fang, and Jie Dong have contributed equally to this work.

Authors and Affiliations

  1. Reproductive Medical Center, Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, 569 Xinyi Road, Baqiao District, Xi’An, Shaanxi Province, China

    Yujing Xiong, Zheng Fang, Jie Dong, Shuqiang Chen, Jiaqin Mao, Wanlin Zhang, Li Hai, Jing Zhou & Xiaohong Wang

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  1. Yujing Xiong
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Contributions

Y.J.X., Z.F., J.D., S.Q.C., and X.H.W. contributed to conceiving and designing the study and to data acquisition. W.L.Z. and L.H. contributed to sample acquisition and article revision. Y.J.X., J.Q.M., J.Z. Z.F., J.D., and X.H.W contributed to data analysis, data interpretation, and drafting and revising the article. All authors approved the final version of the manuscript.

Corresponding author

Correspondence to Xiaohong Wang.

Ethics declarations

Ethics approval and consent to participate

The study was approved by the Tangdu Hospital Ethics Committee (code no. TDLL2018-03-39). Informed consents were acquired from all subjects involved in the study.

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The authors declare no competing interests.

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Supplementary information

ESM 1:

Supplemental Table 1. List of the miRNA/mRNA PCR primers for real-time PCR

ESM 2:

Supplemental Fig. 1 Identification of isolated dNK cells. Flow cytometry was performed to identify the density of CD3-CD56+ dNK cells before and after sorting. Supplemental Fig. 2 Principal component analysis (PCA) plot of DE miRNAs between 4 RPL patients and 5 OP women. Supplemental Fig. 3 Comparison of the expression levels of miR-185-5p among the RPL group and the OP subgroups. (a) Relative expression of miR-185-5p in the OP1 group (patients who maintained a pregnancy without RPL history) and OP2 group (patients who maintained a pregnancy with RPL history). (b) Relative expression of miR-185-5p in the OP1 group and RPL group. (c) Relative expression of miR-185-5p in the OP2 group and RPL group, **** P<0.0001, Mann–Whitney U test was used.

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Xiong, Y., Fang, Z., Dong, J. et al. Maternal circulating exosomal miR-185-5p levels as a predictive biomarker in patients with recurrent pregnancy loss. J Assist Reprod Genet 40, 553–566 (2023). https://doi.org/10.1007/s10815-023-02733-y

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