Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Nov 26, 2024; 16(11): 906-925
Published online Nov 26, 2024. doi: 10.4252/wjsc.v16.i11.906
Effects of miR-214-5p and miR-21-5p in hypoxic endometrial epithelial-cell-derived exosomes on human umbilical cord mesenchymal stem cells
Wan-Yu Zhang, Han-Bi Wang, Cheng-Yan Deng
Wan-Yu Zhang, Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China
Han-Bi Wang, Cheng-Yan Deng, Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Beijing 100730, China
Co-corresponding authors: Han-Bi Wang and Cheng-Yan Deng.
Author contributions: Deng CY conceptualized and designed the study; Wang HB drafted the initial manuscript; Zhang WY collected the data and carried out the initial analyses. Deng CY and Wang HB contribute equally to the study. All authors critically reviewed the manuscript for important intellectual content, approved the final manuscript as submitted, and agree to be accountable for work.
Supported by the National High Level Hospital Clinical Research Funding, No. 2022-PUMCH-B-080 and No. 2022-PUMCH-C-064.
Institutional review board statement: This article does not involve human/animal experiment.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The data used to support the findings of this study are available from the corresponding author upon request.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Cheng-Yan Deng, MD, PhD, Doctor, Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, No. 1 Shuaifuyuan, Dongcheng District, Beijing 100730, China. chydmd@sohu.com
Received: January 25, 2024
Revised: July 24, 2024
Accepted: October 12, 2024
Published online: November 26, 2024
Processing time: 306 Days and 6.2 Hours
Abstract
BACKGROUND

Thin endometrium seriously affects endometrial receptivity, resulting in a significant reduction in embryo implantation, and clinical pregnancy and live birth rates, and there is no gold standard for treatment. The main pathophysiological characteristics of thin endometrium are increased uterine arterial blood flow resistance, angiodysplasia, slow growth of the glandular epithelium, and low expression of vascular endothelial growth factor, resulting in endometrial epithelial cell (EEC) hypoxia and endometrial tissue aplasia. Human umbilical cord mesenchymal stem cells (HucMSCs) promote repair and regeneration of damaged endometrium by secreting microRNA (miRNA)-carrying exosomes. However, the initiation mechanism of HucMSCs to repair thin endometrium has not yet been clarified.

AIM

To determine the role of hypoxic-EEC-derived exosomes in function of HucMSCs and explore the potential mechanism.

METHODS

Exosomes were isolated from normal EECs (EEC-exs) and hypoxia-damaged EECs (EECD-exs), before characterization using Western blotting, nanoparticle-tracking analysis, and transmission electron microscopy. HucMSCs were cocultured with EEC-exs or EECD-exs and differentially expressed miRNAs were determined using sequencing. MiR-21-5p or miR-214-5p inhibitors or miR-21-3p or miR-214-5p mimics were transfected into HucMSCs and treated with a signal transducer and activator of transcription 3 (STAT3) activator or STAT3 inhibitor. HucMSC migration was assessed by Transwell and wound healing assays. Differentiation of HucMSCs into EECs was assessed by detecting markers of stromal lineage (Vimentin and CD13) and epithelial cell lineage (CK19 and CD9) using Western blotting and immunofluorescence. The binding of the miRNAs to potential targets was validated by dual-luciferase reporter assay.

RESULTS

MiR-21-5p and miR-214-5p were lowly expressed in EECD-ex-pretreated HucMSCs. MiR-214-5p and miR-21-5p inhibitors facilitated the migratory and differentiative potentials of HucMSCs. MiR-21-5p and miR-214-5p targeted STAT3 and protein inhibitor of activated STAT3, respectively, and negatively regulated phospho-STAT3. MiR-21-5p- and miR-214-5p-inhibitor-induced promotive effects on HucMSC function were reversed by STAT3 inhibition. MiR-21-5p and miR-214-5p overexpression repressed HucMSC migration and differentiation, while STAT3 activation reversed these effects.

CONCLUSION

Low expression of miR-21-5p/miR-214-5p in hypoxic-EEC-derived exosomes promotes migration and differentiation of HucMSCs into EECs via STAT3 signaling. Exosomal miR-214-5p/miR-21-5p may function as valuable targets for thin endometrium.

Keywords: Endometrial epithelial cells; Exosomes; Human umbilical cord mesenchymal stem cells; MiR-214-5p/miR-21-5p; Signal transducer and activator of transcription 3

Core Tip: Thin endometrium is a primary cause of repeated implantation failure and infertility. In this study, with the help of microRNA sequencing, we investigated the role of hypoxic endometrial epithelial cell (EEC)-derived exosomes in cell function of human umbilical cord mesenchymal stem cells and explored the potential mechanism. Eventually, we found that lowly-expressed miR-21-5p and miR-214-5p in hypoxic EEC-derived exosomes promoted migration and differentiation of human umbilical cord mesenchymal stem cells into EECs via signal transducer and activator of transcription 3 signaling. Exosomal miR-214-5p and miR-21-5p may function as valuable targets for thin endometrium.