miR-644-5p carried by bone mesenchymal stem cell-derived exosomes targets regulation of p53 to inhibit ovarian granulosa cell apoptosis

Abstract

BackgroundThis article aims to reveal the therapeutic effects and potential mechanisms of bone mesenchymal stem cell (BMSC)-derived exosomes on premature ovarian failure (POF).MethodsExosomes were collected from BMSCs and were used to treat cisplatin-induced POF mouse models. Pathological changes of ovarian tissue were detected by using HE staining and by Western blot that detected the expression of apoptosis-related proteins. In cisplatin-induced primary granulosa cell injury, exosomes were co-cultured with the granulosa cells. The apoptosis or viability of granulosa cells was analyzed by flow cytometry or MTT, respectively. In Target scan and microT-CDS databases, an intersection of miRNAs targeting to p53 was found. The expressions of miRNAs in BMSC-derived exosomes were detected by qRT-PCR. Besides, miR-664-5p targeted to p53 of cells was verified by dual-luciferase reporter assay.ResultsBMSC-derived exosomes improved the follicular morphology of POF mice and inhibited the expression of apoptosis-related protein. By co-culture of exosomes and primary granulosa cells, BMSC-derived exosomes repressed cisplatin-induced granulosa cells apoptosis and increased cells viability, while these effects were abrogated after the exosome-containing RNA was degraded by RNase. By Target scan, microT-CDS and qRT-PCR, miR-664-5p was regarded as the dominated RNA in BMSC-derived exosomes. By dual-luciferase reporter assay, miR-664-5p negatively regulated p53 luciferase activity. After shRNA interfering miR-664-5p of BMSC, BMSC-derived exosomes exerted no protective effect on cisplatin-induced granulosa cell apoptosis.ConclusionOur results indicated that miR-644-5p carried by BMSC-derived exosomes inhibited the apoptosis of ovarian granulosa cell by targeting p53 of cells, suggesting that miR-644-5p had the potential to treat POF and restore ovarian function.

In Vitro Generation of Oocyte Like Cells and Their In Vivo Efficacy: How Far We have been Succeeded

“…In this study, hAEC-derived exosomes inhibited granulosa cell apoptosis, protected ovarian vasculature, and maintained the number of primordial follicles in the chemotherapy-damaged ovaries. Similar results were shown by Sun and colleagues [43] who demonstrated the inhibition of ovarian granulosa cells apoptosis by negatively regulating p53 by the administration of BMSC-derived exosomal miR-644-5p into a cisplatin-induced POF mouse model. These results indicated that MSC-derived exosomal miRNA can play a key role in the restoration of ovarian function in chemotherapy-induced animal models.…”

“…Despite having the ability to differentiate into many other cell types under targeted lineage-specific culture conditions, stem cells have shown promising results in curing diseases by their direct insertion into disease models. A number of neonatal and adult sources including bone marrow [43,47,48,55,108,109], endometrial [110], amnion [53,111], amniotic fluid [4], amniotic epithelium [44], menstrual blood [9,51], umbilical cord [56,112], umbilical cord blood [54], umbilical cord vein [52], placenta [49], chorionic plate [50], adipose [113], and skin [46] have shown the restoration of ovarian function in many reproductive disease models. Due to their low immunogenicity, migratory, and homing properties, MSCs have been widely used to restore ovarian functions in POF models.…”

“…In comparison to conventional monolayer (two dimensional: 2D) cultured and control groups, transplanted 3D spheroids not only showed enhanced engraftment efficiency and E 2 levels but also showed the increased expression of folliculogenesis-related markers such as Nanos3, Nobox, and Lhx8. Recently, researchers have used MSC-derived exosomes or microvesicles to evaluate their effect on diseased models [43][44][45]. In a chemotherapy-induced rat ovarian failure study, the transplantation of bone marrow MSCs (BMSCs)-derived exosomes exhibited an increase in the number of basal and sinus follicles, elevated levels of E2 and AMH and reduced levels of FSH and LH in the serum, and it also improved the estrus cycle in rat POF models [55].…”

“…One such cell source with early developmental cell-like properties is that of very small embryonic-like stem cells (VSELs). VSELs have been utilized to derive OLCs in vitro, and micro-RNAs (miRNA) from VSELs have been shown to have the peculiar ability to treat female reproductive disorders like premature ovarian failure (POF) and premature ovarian insufficiency (POI) [43,44]. Despite these wonderful properties, VSELs have also been found to survive chemotherapeutic applications, which makes them a more valuable cell source [45].…”

Mesenchymal Stem Cells as a Bio Organ for Treatment of Female Infertility

“…Exosomes are important vehicles for intercellular communication, and as such, are critical for many physiological functions. Recent studies indicated that exosomal miRNAs can regulate intercellular signal transduction and control molecular activities in different diseases [ 60 , 61 , 62 , 63 ]. Two studies have demonstrated a role of miRNAs carried by BMSCs-derived exosomes in promoting the recovery of ovarian function in a POF animal model [ 61 , 62 ].…”