Endometrial transcriptome and PGR cistrome in cycling fertile women [ChIP-seq]
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ABSTRACT: We report the transcriptomic profile and PGR cistromic profile for endometrial biopsies obtained from fertile women during the proliferative and mid-secretory phase of the menstrual cycle
Project description:We report the transcriptomic profile and PGR cistromic profile for endometrial biopsies obtained from fertile women during the proliferative and mid-secretory phase of the menstrual cycle
Project description:Whole genome expression analyses of autologous, paired eutopic and ectopic endometrial samples obtained during proliferative and secretory phases of menstrual cycles from eighteen (n=18) fertile women suffering from confirmed stage 3 (moderate) and stage 4 (severe) ovarian endometriosis were performed using whole human genome oligo microarray Agilent paltform (Cat. No. G4112F). In the present study, genome-wide expression analysis of autologous, paired eutopic and ectopic endometrial samples obtained during proliferative (n=13) and secretory (n=5) phases of menstrual cycle from fertile women (n=18) suffering from moderate (stage 3; n=8) or severe (stage 4; n=10) endometrioma was performed by using Agilent single color oligo microarray platform (G4112, 4X44K). Thus eighteen (18) eutopic (shown as EU) and eighteen (18) ectopic (shown as EC) samples from eighteen (18) subjects with confirmed menstrual phase (proliferative and secretory) and severity stages (stage 3 and stage 4) were studied.
Project description:Whole genome expression analyses of autologous, paired eutopic and ectopic endometrial samples obtained during proliferative and secretory phases of menstrual cycles from eighteen (n=18) fertile women suffering from confirmed stage 3 (moderate) and stage 4 (severe) ovarian endometriosis were performed using whole human genome oligo microarray Agilent paltform (Cat. No. G4112F).
Project description:We report the genome-wide binding sites of PGR-A and PGR-B at 2h of in vitro differentiation of human endometrial stromal cells that express either PGR-A or PGR-B. Progesterone, acting through the progesterone receptors (PGRs), is one of the most critical regulators of endometrial differentiation, known as decidualization, which is a key step toward the establishment of pregnancy. Yet a long-standing unresolved issue in uterine biology is the precise roles played by the major PGR isoforms, PGR-A and PGR-B, during decidualization in the human. Our approach, expressing PGR-A and PGR-B individually after silencing endogenous PGRs in human endometrial stromal cells (HESC), enabled the analysis of the roles of these isoforms separately as well as jointly by ChIP-seq and gene-expression analysis. In order to study the cistromes of PGR-A and PGR-B at 2h of in vitro differentiation of human endometrial stromal cells, we generated primary cultures of human endometrial stromal cells expressing flag tagged PGR-A and PGR-B individually after silencing endogenous PGRs. Input DNA was used as the reference sample.
Project description:Endometrial receptivity is imperative to achieving pregnancy in humans. A disruption in the development of endometrial receptivity is responsible for recurrent implantation failures (RIF) of endometrial origin. To further understand the molecular mechanisms behind the endometrial receptivity process, we used the 8-plex isobaric tag for relative and absolute quantitation (iTRAQ) method to compare and quantify the proteomes from endometrial biopsies of three different endometrial statuses (fertile women, IUD carriers and RIF patients). Overall, iTRAQ allowed to identify 1,889 non-redundant proteins. Of these, 188 were differentially expressed proteins (DEP) (p-value < 0.05) among the three endometrial groups. Pairwise comparisons revealed 133 significant DEP in fertile vs. IUD carriers and 158 DEP in RIF vs. IUD carriers. However, no DEP were identified between fertile and RIF patients. Western blot validation of three DEP involved in endometrial receptivity (Plastin 2, Lactotrasferrin, and Lysozyme) confirmed our iTRAQ results. Moreover, functional KEGG enrichment revealed that complement and coagulation cascades and peroxisome were the two most significant pathways for the RIF vs. IUD comparison and ribosome and spliceosome for the fertile vs. IUD comparison, as possible important pathways involved in the endometrial receptivity acquisition. Our findings confirm that an IUD introduces numerous changes in the endometrial protein profile when compared to fertile and RIF endometria, revealing some key proteins involved in endometrial receptivity. The lack of DEP between fertile and RIF patient endometria suggest either that idiopathic RIF may not have an endometrial origin, with other as-yet-unknown factors involved.
Project description:Progesterone receptor (PGR) is essential for various functions in the female reproductive tract, especially ovulation in the ovary. PGR consists of two main isoforms, PGR-A and PGR-B, that possess different expression patterns and are responsible for various unique roles in different tissue contexts. While both are present in the reproductive tract, PGR-A is the only one known to be essential for ovulation while PGR-B plays a lesser role in the reproductive tract. However, the specific roles of PGR isoforms on gene expression in peri-ovulatory granulosa cells have not been previously investigated in detail. In this study, we identified the unique role of PGR-A and PGR-B in peri-ovulatory granulosa cells through RNA-seq of knockout mouse models for total PGR (PGR KO) or specific isoforms (PGR-A KO and PGR-B KO)