Abstract
Recurrent pregnancy loss is a common obstetric complication affecting approximately 1–2% of reproductive population worldwide, but the precise causes for approximately a half of such patients remain unexplained. In this study, we compared the expression profiles of messenger RNA (mRNA), long non-coding RNA (lncRNA), microRNA (miRNA), and circular RNA (circRNA) in villi tissues from patients with unexplained recurrent pregnancy loss (URPL) and elective termination of pregnancy (ETP) using whole-transcriptome sequencing. A number of differentially expressed RNAs were confirmed by real-time PCR analysis. As a result, we identified a total of 1,703 mRNAs, 798 lncRNAs, 199 miRNAs, and 163 circRNAs that were significantly differentially expressed between villi tissues from URPL and ETP. The data of real-time PCR were consistent with those of the sequencing results. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the majority of differentially expressed mRNAs and target genes of ncRNAs were associated with focal adhesion, extracellular matrix-receptor interaction, and the PI3K-Akt signaling pathway. Additionally, two co-expression networks (lncRNA–miRNA–mRNA and lncRNA-circRNA–miRNA–mRNA) were constructed based on the correlation analysis between the differentially expressed RNAs. Taken together, this study provides a large number of valuable candidates for elucidating regulatory mechanisms of ncRNAs, which may ultimately assist in understanding the pathogenesis of URPL.
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Data availability
All RNA-Seq data have been submitted to the NCBI Sequence Read Archive with BioProject accession number PRJNA760763. Further inquiries can be directed to the corresponding authors.
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Acknowledgements
We would like to thank Mr. Song Shi from Shanghai OE Biotech (Shanghai, China) for his assistant in data analysis.
Funding
This study was supported by grants from the National Natural Science Foundation of China (Nos. 81801445 and 81770236) and the Nan**g Health Science and Technology Development Project-Key Medical Science and Technology Development Project (No. ZKX18045).
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ZXu, JT, and QX conceived and designed the study. YW and QC performed the majority of the experiments and analyzed the data. Z**, YL, and LM collected clinical samples. RZ gave technical support and conceptual advice. YW and QC wrote the manuscript. ZXu and JT revised the manuscript. All authors approved the final manuscript.
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Written informed consent was obtained from all patients, and this study was approved by the Institutional Review Board of Nan**g Maternity and Child Health Care Hospital (No. 2021KY004).
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The authors declare no competing interests.
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Wang, Y., Cheng, Q., **a, Z. et al. Whole-transcriptome sequencing identifies key mRNAs, miRNAs, lncRNAs, and circRNAs associated with unexplained recurrent pregnancy loss. Cell Tissue Res 389, 129–143 (2022). https://doi.org/10.1007/s00441-022-03632-x
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DOI: https://doi.org/10.1007/s00441-022-03632-x