Next Generation Sequencing Analysis of exosomes in follicle fluid of PCOS/non-PCOS patients
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ABSTRACT:
PROVIDER: PRJNA607262 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11100948_1.fastq.gz | Fastqsanger.gz | |||
| SRR11100948_2.fastq.gz | Fastqsanger.gz | |||
| SRR11100949_1.fastq.gz | Fastqsanger.gz | |||
| SRR11100949_2.fastq.gz | Fastqsanger.gz | |||
| SRR11100950_1.fastq.gz | Fastqsanger.gz |
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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: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:Aberrant expression of long noncoding RNAs in exosomes in follicle fluid from PCOS patients
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Project description:Preparation of exosomes isolated from semen contain a substantial amount of RNA, mostly from 20 to 100 nucleotides in length. We sequenced separately 20-40 and 40-100 nucleotide fractions of RNA from exosomes isolated from semenal fluid from six healthy donors. We found various classes of small non-coding RNA, including mature microRNA and piwi-RNA, as well as abundant Y RNAs and tRNAs present in both full length and fragmented forms. Specific RNAs were consistently present in all donors. For example, fifteen (of ~2,600 known) microRNAs constituted over 80% of mature microRNA in SE. Additionally, tRNA fragments were strongly enriched for 5M-bM-^@M-^Y-ends of 18-19 or 30-34 nucleotides in length. Size-fractionated small RNA profiles from exosomes isolated from the seminal fluid of six healthy donors
2014-06-04 | E-GEOD-56076 | biostudies-arrayexpress
Project description:Exosomes have recently been shown to play a key role in cell-to-cell communication through delivery of various functional content, including microRNAs (miRNAs). We investigated the potential roles of exosomal miRNA derived intrafollicular cells in polycystic ovary syndrome (PCOS). Using microarray profiling, a total of 492 miRNAs and 220 miRNAs were found in follicular fluid-derived exosomes and serum-derived exosomes, respectively, in PCOS and non-PCOS females. By excluding miRNAs existing in serum-derived exosomes, we found 179 miRNAs which were specifically expressed in follicular fluid-derived exosomes both in PCOS and non-PCOS females. Using microarray profiling, a total of 492 miRNAs and 220 miRNAs were found in follicular fluid-derived exosomes and serum-derived exosomes, respectively, in PCOS and non-PCOS females. By excluding miRNAs existing in serum-derived exosomes, we found 179 miRNAs which were specifically expressed in follicular fluid-derived exosomes both in PCOS and non-PCOS females.
2022-02-01 | GSE145318 | GEO
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: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 long non-coding RNAs (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 1,253 upregulated and 613 downregulated lncRNAs from a total of 1,866 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 pathways related to endocytosis, the Hippo, the MAPK, and HTLV-1 infection. 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.
| S-EPMC7925891 | biostudies-literature
Project description:Polycystic ovary syndrome (PCOS), the most common cause of anovulatory infertility, is characterized by increased ovarian androgen production, arrested follicle development, and is frequently associated with insulin resistance. These PCOS phenotypes are associated with exaggerated ovarian responsiveness to FSH and increased pregnancy loss. To examine whether the perturbations in follicle growth and the intrafollicular environment affects development of the mature PCOS oocyte, genes that are differentially expressed in PCOS compared to normal oocytes were defined using microarray analysis. This analysis detected approximately 8000 transcripts. Hierarchical clustering and principal component analysis revealed differences in global gene expression profiles between normal and PCOS oocytes. 374 genes had a statistically-significant increase or decrease in mRNA abundance in PCOS oocytes. A subset of these genes was associated with chromosome alignment and segregation during mitosis and/or meiosis, suggesting that increased mRNAs for these proteins may negatively affect oocyte maturation and/or early embryonic development. Of the 374 differentially expressed genes, 68 contained putative androgen receptor, retinoic acid receptor, and/or peroxisome proliferating receptor gamma binding sites, including 9 of the genes involved in chromosome alignment and segregation. These analyses demonstrated that normal and PCOS oocytes that are morphologically indistinguishable and of high quality exhibit different gene expression profiles. Furthermore, altered mRNA levels in the PCOS oocyte may contribute to defects in meiosis and/or mitosis which might impair oocyte competence for early development and therefore contribute to poor pregnancy outcome in PCOS. Keywords: disease state analysis
2007-06-05 | GSE5850 | GEO
Project description:Preparation of exosomes isolated from semen contain a substantial amount of RNA, mostly from 20 to 100 nucleotides in length. We sequenced separately 20-40 and 40-100 nucleotide fractions of RNA from exosomes isolated from semenal fluid from six healthy donors. We found various classes of small non-coding RNA, including mature microRNA and piwi-RNA, as well as abundant Y RNAs and tRNAs present in both full length and fragmented forms. Specific RNAs were consistently present in all donors. For example, fifteen (of ~2,600 known) microRNAs constituted over 80% of mature microRNA in SE. Additionally, tRNA fragments were strongly enriched for 5’-ends of 18-19 or 30-34 nucleotides in length.
2014-06-04 | GSE56076 | GEO
Project description:Exosomes and microvesicles (i.e., extracellular vesicles; EVs) have been identified within ovarian follicular fluid, and recent evidence suggests that EVs are able to elicit profound effects on ovarian cell function. While existence of miRNA within EVs has been reported, it remains unknown if EV size and concentration as well as their cargos (i.e., proteins and RNA) change during antral follicle growth. Extracellular vesicles isolated from follicular fluid of small, medium and large bovine follicles were similar in size, while concentration of EVs decreased progressively as follicle size increased. Electron microscopy indicated a highly purified population of the lipid bilayer enclosed vesicles that were enriched in exosome biomarkers including CD81 and Alix. Small RNA sequencing identified a large number of known and novel miRNAs that changed in the EVs of different size follicles. Ingenuity Pathway Analysis (IPA) indicated that miRNA abundant in small follicle EV preparations were associated with cell proliferation pathways, while those miRNA abundant in large follicle preparations were related to inflammatory response pathways. These studies are the first to demonstrate that EVs change in their levels and makeup during antral follicle development and point to the potential for a unique vesicle-mediated cell-to-cell communication network within the ovarian follicle. Examination of small RNA population in bovine follicular fluid extracellular vesicles isolated from antral follicles
2016-02-09 | E-GEOD-74879 | biostudies-arrayexpress