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Improving Granulosa Cell Function in Premature Ovarian Failure with Umbilical Cord Mesenchymal Stromal Cell Exosome-Derived hsa_circ_0002021

  • Original Article
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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

The therapeutic potential of exosomes from human umbilical cord mesenchymal stem cells (HUMSCs-Exo) for delivering specific circular RNAs (circRNAs) in treating premature ovarian failure (POF) is not well understood. This study aimed to explore the efficacy of HUMSCs-Exo in delivering hsa_circ_0002021 for POF treatment, focusing on its effects on granulosa cell (GC) senescence and ovarian function.

Methods:

Bioinformatic analysis was conducted on circRNA profiles using the GSE97193 dataset from GEO, targeting granulosa cells from varied age groups. To simulate granulosa cell senescence, KGN cells were treated with cyclophosphamide (CTX). HUMSCs were transfected with pcDNA 3.1 vectors to overexpress hsa_circ_0002021, and the HUMSCs-Exo secreted were isolated. These exosomes were characterized by transmission electron microscopy (TEM) and Western blotting to confirm exosomal markers CD9 and CD63. Co-culture of these exosomes with CTX-treated KGN cells was performed to assess β-galactosidase activity, oxidative stress markers, ROS levels, and apoptosis via flow cytometry. Interaction between hsa_circ_0002021, microRNA-125a-5p (miR-125a-5p), and cyclin-dependent kinase 6 (CDK6) was investigated using dual-luciferase assays and RNA immunoprecipitation (RIP). A POF mouse model was induced with CTX, treated with HUMSCs-Exo, and analyzed histologically and via immunofluorescence staining. Gene expression was quantified using RT-qPCR and Western blot.

Results:

hsa_circ_0002021 was under expressed in both in vivo and in vitro POF models and was effectively delivered by HUMSCs-Exo to KGN cells, showing a capability to reduce GC senescence. Overexpression of hsa_circ_0002021 in HUMSCs-Exo significantly enhanced these anti-senescence effects. This circRNA acts as a competitive adsorbent of miR-125a-5p, regulating CDK6 expression, which is crucial in modulating cell cycle and apoptosis. Enhanced expression of hsa_circ_0002021 in HUMSCs-Exo ameliorated GC senescence in vitro and improved ovarian function in POF models by modulating oxidative stress and cellular senescence markers.

Conclusion:

This study confirms that hsa_circ_0002021, when delivered through HUMSCs-Exo, can significantly mitigate GC senescence and restore ovarian function in POF models. These findings provide new insights into the molecular mechanisms of POF and highlight the therapeutic potential of circRNA-enriched exosomes in treating ovarian aging and dysfunction.

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Data availability

Data is available from the corresponding author on request.

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Acknowledgements

Sichuan Woman and Child Health Association 2023 Maternal and Child Medical Science and Technology Innovation Project Important Research Subject (No. FXZD03).

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Authors and Affiliations

Authors

Contributions

GY conceived and designed the evaluation and drafted the manuscript. BZ, Mei Xu and MW participated in designing the evaluation, performed parts of the statistical analysis and helped to draft the manuscript. JL, ZL and YHC re-evaluated the data, revised the manuscript and performed the statistical analysis. QH, GPH and HYH collected the data, interpreted them and revised the manuscript. GY re-analyzed the statistical data and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to HaiYan Hu.

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The authors declare that they have no conflict of interest.

Ethical statement

The animal experiment research protocol was approved by the Ethics Committee of Zigong Maternity and Child Health Care Hospital (No.2020FX0311) and performed in accordance with the “Guidelines for the care and use of experimental animals.”

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Yang, G., Zhang, B., Xu, M. et al. Improving Granulosa Cell Function in Premature Ovarian Failure with Umbilical Cord Mesenchymal Stromal Cell Exosome-Derived hsa_circ_0002021. Tissue Eng Regen Med (2024). https://doi.org/10.1007/s13770-024-00652-2

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  • DOI: https://doi.org/10.1007/s13770-024-00652-2

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