Human Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles Carrying MicroRNA-29a Improves Ovarian Function of Mice with Primary Ovarian Insufficiency by Targeting HMG-Box Transcription Factor/Wnt/β-Catenin Signaling

Abstract

Background. Primary ovarian insufficiency (POI) is a female disease characterized by ovarian function loss under 40 years old. Transplantation of exosomes is an encouraging regenerative medicine method that has the potential for restoring ovarian functions post-POI with high efficiency. Therefore, we investigate the therapeutic efficacy and potential mechanisms of human umbilical cord mesenchymal stem cell- (UCMSC-) derived exosomes on ovarian dysfunction post-POI. Methods. The model of POI was established by intraperitoneal injection with 5 mg/kg cisplatin. The mouse ovarian function was detected by measuring the levels of anti-Mullerian hormone, follicle-stimulating hormone, and estradiol and detecting the morphological changes. For in vitro experiments, the characterization and identification of UCMSCs and UCMSC-derived exosomes were done by observation of morphologies and flow cytometry. To exclude the interference effect of nonspecific precipitation substances, UCMSCs were treated with RNase A or RNase A in combination with Triton X-100. Granulosa cell (GC) identification was performed using immunofluorescence. GC proliferation and viability were assessed using 5-ethynyl-2 ′ -deoxyuridine (EdU) assays and Cell Counting Kit-8 (CCK-8), and GC apoptosis was calculated by flow cytometry. Gene expression and protein levels were evaluated using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting. The binding relationship between miR-29a and HMG-box transcription factor (HBP1) was verified by luciferase reporter assays. Results. In vitro, the human UCMSC-derived exosomes carrying miR-29a upregulation promoted the proliferation of GCs and suppressed their apoptosis. In vivo, miR-29a upregulation reserved the mature follicles and restored the ovarian functions. miR-29a targeted HBP1 and negatively regulated its expression. HBP1 upregulation rescued the miR-29a upregulation-induced inhibition in GC apoptosis and inactivated the Wnt/β-catenin pathway. Conclusion. The exosomal miR-29a derived from human UCMSCs improves the ovarian function by targeting HBP1 and activating the Wnt/β-catenin pathway.

β-Catenin regulates ovarian granulosa cell cycle and proliferation in laying hens by interacting with TCF4

“…Earlier studies report several factors regulating GC proliferation and differentiation, most of which are related to the Wnt/β-catenin signaling pathway. For example, miR-23a-3p decreased high mobility group AT-hook 2 ( HMGA2 ) expression, blocking the Wnt/β-catenin pathway, thereby depressing viability and facilitating apoptosis of GCs ( Wei et al, 2023 ); miR-29a promotes proliferation and inhibits apoptosis of GCs by targeting HMG-box transcription factor 1 ( HBP1 ) and activating the Wnt/β-catenin pathway ( Gao et al, 2022 ); SMAD family member 4 ( SMAD4 ) inhibits granulosa cell apoptosis ( Du et al, 2020 b) and bone morphogenetic protein-15 ( BMP15 ) in granulosa cells can promote follicular selection in hens ( Stephens and Johnson, 2016 ). The major members of the Wnt family are secreted glycoproteins that play numerous roles in regulating embryonic development, cell growth and differentiation, and cell fate ( Boyer et al, 2010 a, b ; Richards, 2018 ).…”

Stem cell-derived extracellular vesicles: A novel and potential remedy for primary ovarian insufficiency

“…There are round or oval membranous vesicles, which can be aggregated and distributed, and their membrane structure is clearly defined ( Vohra et al, 2020 ). hUCMSCs-EVs express the EVs-specific four-transmembrane protein markers CD9, CD63, CD81 and tumor susceptibility gene 101 protein (TSG101), heat shock protein70 (HSP70), and the multivesicular biosynthesis-related protein ALIX ( Sun et al, 2017 ; Yang et al, 2019 ; Zhang et al, 2020 ; Deng et al, 2021 ; Cai et al, 2022 ; Gao et al, 2022 ; Qu et al, 2022 ). In 2017, hUCMSCs- EVs were first described for application to POI by Sun et al (2017) .…”

“…Furthermore, the EVs-mediated transfer of miRNA is an important way by which stem cell function. Some miRNA including miR-126-3p, miR-21, miR-29a, and miR-17-5P carried by hUCMSCs-EVs could play a role at the post-transcriptional level ( Ding et al, 2020 ; Cai et al, 2022 ; Gao et al, 2022 ; Qu et al, 2022 ). By binding to the 3′UTR of target genes, these miRNAs inhibit the expression of particular molecules to inhibit reactive oxygen species (ROS) production and apoptosis and promote cell survival, proliferation, and angiogenesis in GCs.…”

Biological therapies for premature ovarian insufficiency: what is the evidence?

“…This understanding has stimulated research in order to investigate potential disease-modifying role of micro-RNAs in chemotherapy-induced mice models of POI, with so far, promising results. For example, miR-17-5p, derived from human UC-MSCs-exosomes, promotes the proliferation of damaged granulosa and ovarian cells and decreases oxidative stress, via inhibition of sirtuin 7 (SIRT7) expression (110). In addition, miR-29α promotes the proliferation of granulosa cells and suppresses their apoptosis, reserves the existing mature follicles and restores ovarian function, via targeting wingless-related integration (Wnt)/β-catenin pathway (111).…”

“…For example, miR-17-5p, derived from human UC-MSCs-exosomes, promotes the proliferation of damaged granulosa and ovarian cells and decreases oxidative stress, via inhibition of sirtuin 7 (SIRT7) expression ( 110 ). In addition, miR-29α promotes the proliferation of granulosa cells and suppresses their apoptosis, reserves the existing mature follicles and restores ovarian function, via targeting wingless-related integration (Wnt)/β-catenin pathway ( 111 ). Up to now, there are no clinical data about the use of microRNAs in patients with POI.…”