Identification of exosomal miRNAs specifically derived from intrafollicular cells involved in polycystic ovary syndrome
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE145318 | GEO | 2022/02/01
REPOSITORIES: GEO
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Project description:Polycystic ovary syndrome (PCOS) is a complex endocrinopathy affecting reproductive aged women, whose etiology has not been fully understood yet. The follicular growth is arrested at preantral stage leading to cyst formation, consequently resulting in anovulatory infertility in these women. As follicular fluid provides the microenvironment for the growing oocyte, molecular profiling of the fluid may provide unique information about pathophysiology associated with follicular development in PCOS. Modification with oligosaccharide chains are known to influence functions of several secreted proteins and these glycoproteins also play a role in disease pathology. The glycoproteomic profile of follicular fluid of PCOS has not been explored in PCOS yet. In the present study, we performed comparative glycoproteomic analysis by first enriching glycoproteins using three different lectins viz. concanavalin A, wheat germ agglutinin and Jacalin from follicular fluid of women with PCOS and controls undergoing in vitro fertilization. Peptides generated by trypsin digestion were labeled with isobaric tags for relative and absolute quantification reagents and analyzed by liquid chromatography tandem mass spectrometry. We identified 10 differentially expressed glycoproteins, in the follicular fluid of women with PCOS compared to control. Two important differentially expressed proteins- SERPINA1 and ITIH4, were consistently upregulated and downregulated respectively, upon validation by Western blotting in follicular fluid and real-time polymerase chain reaction in granulosa cells. These proteins play a role in angiogenesis and extracellular matrix stabilization which are vital for follicle maturation. In conclusion, comparative glycoprotein profiling of follicular fluid from women with PCOS and controls revealed altered expression of proteins which may contribute to defects in follicle development in PCOS pathophysiology.
2019-03-22 | PXD012731 | Pride
Project description:Purpose: the goals of this study are to identify the transcriptome profiling of exosome derived from follicle fluid of PCOS patients by next-generation sequencing (NGS). Methods:the transcriptome profiling of exosome derived from follicular fluid of five PCOS and five control patients were generated by deep sequencing, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: the transcriptome profiling including circRNA, lncRNA and mRNA were comparatively analyzed in the two groups of exosomes (exo-PCOS and exo-control). We focus on the circRNAs profile for further analysis. A circRNA candidate was considered to be differentially expressed if its levels between both experiment groups showed a statistically significant difference (p<0.05) of at least two-fold. Based on this criterion, we identified 16 differentially expressed circRNAs (5 up-regulated and 11 down-regulated circRNAs) between exo-PCOS and exo-control, including 3 annotated exonic circRNAs and 13 novel ones. Clustering analysis based on the differentially expressed circRNAs clustered the exo-PCOS and the exo-control groups (control). Conclusion: the circRNA expression profile of exosome derived from follicular fluid of five PCOS and five control patients were determined by RNA sequencing technology. 16 circRNAs showed significantly different expression in the exosome of PCOS FF. GO and KEGG pathway analysis indicated that the parental genes of the candidate circRNAs were involved in ovarian steroidogenesis, aldosterone synthesis and secretion, thyroid hormone signaling pathway, ubiquitin mediated proteolysis, Jak-STAT signaling pathway, and endocytosis pathway.
2020-09-09 | GSE145461 | GEO
Project description:The follicular fluid (FF) fills the interior of ovarian antral follicle and provides the microenvironment for oocyte growth and acquisition of its competence to ovulate and latter support early embryo development. The FF is derived from both blood plasma and secretion of different types of follicular cells. It contains also extracellular vesicles (EVs), including exosomes, small membrane-coated EVs with 30-150 nm in diameter, which participate in cell-to cell communication and signaling by transferring their cargo of different types of RNAs, proteins and lipids into the oocyte or follicular cells. To date most studies have focused on studying the ffEVs miRNAs cargo and showing that miRNAs can influence oocyte competence and further embryo development. However, ffEVs protein cargo, which could have a direct contribution after being uptake by the oocyte or follicular cells have been less studies.
2021-08-17 | PXD020508 | Pride
Project description:Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disease characterized by persistent anovulation and hyperandrogenism, affecting approximately 8%-10% of women of childbearing age and occupying an important position in the etiology of infertility. There is increasing evidence that lncRNAs are involved in the development of PCOS, but the potential regulatory mechanism is still unclear. This study performed high-throughput lncRNA sequencing of follicular fluid exosomes in non-PCOS infertility patients and PCOS infertility patients. The sequencing results led to the identification of 1253 upregulated and 613 downregulated lncRNAs from a total of 1866 detected candidates. There was no significant difference between the PCOS patients and non-PCOS patients in body mass index (BMI) or the fasting blood glucose (FBG) level. However, luteinizing hormone (LH), estradiol (E2), testosterone (T), serum prolactin (PRL) and anti-Mullerian hormone (AMH) levels were clearly upregulated in PCOS patients compared to those in non-PCOS patients. There was also an increase in LH/FSH (>2) in the PCOS patients. Functional analysis showed that the functions of lncRNAs were related to adenine metabolism, adenine biosynthesis and cytidine triphosphate biosynthesis. These results suggest that lncRNAs may play an important role in the pathogenesis of PCOS and may be potential targets for the diagnosis and treatment of PCOS.
2021-03-11 | GSE159466 | GEO
Project description:Proteomic analysis of follicular fluid exosomes derived from SYFs and ASYFs in geese
2024-08-10 | PXD049994 | Pride
Project description:Kashin-Beck disease (KBD) is an endemic, chronic degenerative joint disease in China. Exosomes miRNAs, as signaling molecules in intercellular communication, can transfer specific biological martials into target cell to regulate the their function and might participate in the pathogenesis of KBD. We isolated serum and chondrocytes-derived exosomes, miRNA sequencing revealed exosomes miRNA profiles and differentially expressed miRNAs (DE-miRNAs) were identified. The target genes were predicted of known and novel DE-miRNAs with TargetScan 5.0 and miRanda 3.3a database. Single-cell RNA sequencing (scRNA-seq) was performed to identify chondrocyte clusters and their gene signatures in KBD. And we performed comparative analysis between the serum and chondrocytes-derived exosomes DE-miRNA target genes and differentially expressed genes of each cell clusters.A total of 20 DE-miRNAs were identified in serum-derived exosomes. In the miRNA expression of chondrocytes-derived exosomes, 53 DE-miRNAs were identified. 16063 predicted targets were identified as the target genes in the serum-derived exosomes, 57316 predicted targets were identified as the target genes in the chondrocytes-derived exosomes. Seven clusters were labelled by cell type according to the expression of previously described markers. 315 common genes were found among serum/chondrocytes-derived exosomes DE-miRNA target genes and DEGs identified by scRNA-seq analysis. We firstly integratly analyzed the serum and chondrocytes exosomes miRNA with single-cell RNA sequencing (scRNA-seq) data of KBD chondrocyte, the results showed that DE-miRNAs in exosomes might play a potential role in regulating genes expression in different KBD chondrocytes clusters by exosomes mediating cell-cell communications functions, which could improve the new diagnosis and treatment methods for KBD.
2023-11-01 | GSE246097 | GEO
Project description:Currently, micro RNAs (miRNAs) constitute a promising models for cell-to-cell communication since they are transferred between cells to execute essential roles in many processes. Their structure and size, in addition to various transport mechanisms, allow them to remain stable within various biological fluids, surviving in extremely adverse conditions, including low pH, boiling, and freezing. The presence of miRNAs in reproductive fluids, such as follicular, uterine and seminal fluid, indicate potential roles in the reproductive system. These extracellular miRNAs can be transported by lipoproteins (both HDL and LDL) or other proteins, including Argonaute2 (AGO2) and nucleophosmin1 (NPM1). Another transport system is mediated by extracellular vesicles (EVs), such as apoptotic bodies, microvesicles (MVs) and/or exosome-like vesicles. EVs protect miRNAs from degradation and contribute to their stability within biological fluids. Furthermore, EVs can transport a wide range of components packaged in a selective way. For instance, Squadrito et al. (2014), used macrophages and endothelial cells to demonstrate that the sorting of miRNAs into EVs for heterotrophic cell communication is altered by both, the presence of target transcripts and the self-presence of the respectively miRNA. In addition, the signature of miRNAs found in the exosomes significantly differed for those detected in the parent cells. To date, mechanisms controlling the specific loading of miRNAs into exosomes remain unclear. Indeed, several mechanisms may govern exosome sorting of specific subsets of miRNAs. MiRNA sorting appears to be influenced by different pathways and molecules in different cell types and tissues, and miRNAs contain well defined motifs (i.e., EXOmotifs), that direct the miRNA allocation into exosomes before delivery into recipient cells. A recent study showed that this RNA sequence can be recognized by the sumoylated form of the heterogeneous ribonucleoprotein A2B1 (hnRNPA2B1). Moreover, a terminal 30 nucleotide addition in miRNAs affects their selective sorting in B cells. Another hypothesis suggests that RNAs are selectively sorted depending on the differential affinity of RNA motifs towards the raft-like region of the cytoplasmic surface of microvesicular body (MVB) limiting membranes. Current data indicate that cells can communicate with each other through the transfer of miRNA-loaded exosomes. For example, monocyte-derived exosomes deliver miR-150 to endothelial cells and enhance endothelial cell migration by reducing c-myb expression. The miRNA content of exosomes plays a critical role in such cell-to-cell communication and determines the fate of recipient cells. Thus, exosomes derived from the bone marrow mesenchymal stromal cells of myeloma patients promote tumor growth depending on the content of miR-15a in exosomes. Our group has described a novel cell-to-cell communication mechanism involving the delivery of endometrial miRNAs from the maternal endometrium to the trophectoderm cells of preimplantation embryos. Specifically, in B6C3 derived mouse embryos, we found EV-associated and free miR-30d to cause overexpression of genes involved in embryonic adhesion processes, including Itb3, Itga7 and Cdh5. Furthermore, supplementing murine embryos with miR-30d significantly improved embryo adhesion, suggesting that external miRNAs may have a functional role as transcriptomic modifiers of preimplantation embryos. Based on profiling of miRNAs in endometrial fluid, maternally-derived miRNAs are present within EVs in the uterine microenvironment. The internalization of maternally-derived exosomes has been visualized, but the mechanism by which miR-30d becomes incorporated into exosomes remains unknown. The present study aimed to elucidate the underlying mechanism of hsa-miR-30d transfer from human endometrial epithelial cells (hEECs) to the interior of exosomes and eventually to early-stage blastocysts, using a mir-30d knockout murine mode.
2018-06-20 | PXD008773 | Pride
Project description:The aim of this study was to analyze and compare the differential expression of peptides within the follicular fluid of polycystic ovary syndrome (PCOS) patients versus normal women by using peptidomics techniques. The underlying mechanisms involved in PCOS pathogenesis will be explored, together with screening and identification of potential functional peptides via bioinformatics analysis.
2023-10-27 | PXD045753 | Pride