Project description:Clinical treatment protocols for infertility with in vitro fertilization-embryo transfer (IVF-ET) provide a unique opportunity to assess the human vaginal microbiome in defined hormonal milieu. Herein, we have investigated the association of circulating ovarian-derived estradiol (E2) and progesterone (P4) concentrations to the vaginal microbiome. Thirty IVF-ET patients were enrolled in this study, after informed consent. Blood was drawn at four time points during the IVF-ET procedure. In addition, if a pregnancy resulted, blood was drawn at 4-to-6 weeks of gestation. The serum concentrations of E2 and P4 were measured. Vaginal swabs were obtained in different hormonal milieu. Two independent genome-based technologies (and the second assayed in two different ways) were employed to identify the vaginal microbes. The vaginal microbiome underwent a transition with a decrease in E2 (and/or a decrease in P4). Novel bacteria were found in the vagina of 33% of the women undergoing IVF-ET. Our approach has enabled the discovery of novel, previously unidentified bacterial species in the human vagina in different hormonal milieu. While the relationship of hormone concentration and vaginal microbes was found to be complex, the data support a shift in the microbiome of the human vagina during IVF-ET therapy using standard protocols. The data also set the foundation for further studies examining correlations between IVF-ET outcome and the vaginal microbiome within a larger study population.
Project description:The aim of the study was to investigate if the use of a 12 weeks liraglutide treatment change the endometrial gene expression during the implantation window in infertile obese women with polycystic ovary syndrome (PCOS) undergoing in-vitro fertilization (IVF). This was a cross-sectional study involving treated and non-treated subjects. Endometrial biop-sies were collected from 20 infertile women with PCOS and BMI ≥30 kg/m2 before the IVF procedure. Endometrium transcriptome of 10 participants who had been pretreated with low-dose liraglutide 1.2 mg QD for 12 weeks and achieved at least a 5% reduction in body weight was compared to endometrium transcriptome of 10 treatment-naive participants who had a sta-ble body weight over the last 12 weeks. Next-generation sequencing was conducted to analyze RNA samples. The resulting data were processed to discern key canonical pathways and pre-dict activations or inhibitions. Gene networks were constructed based on established published associations.
Project description:We analysed 227 infertile couples to determine the frequency of pregnancies occurring without any specific treatment. Pregnancy occurred in 44 of 104 (42%) couples who were actually treated and 43 of 123 (35%) couples who formed treatment independent pregnancy group. Treatment independent pregnancy group accounted for 43 of total 87 pregnancies occurring in this study. The study demonstrates that the potential for spontaneous cure for infertility is high and many patients respond favourably to just attending infertility clinic rather than specific treatment.
Project description:The seminal vesicles are male accessory sex glands that contribute the bulk of the seminal plasma in which mammalian spermatozoa are bathed during ejaculation. In addition to helping convey sperm through the ejaculatory duct, seminal vesicle secretions support sperm survival after ejaculation and influence the female reproductive tract to promote receptivity to pregnancy. Despite its biological importance, there is limited understanding of seminal vesicle fluid (SVF) composition with only 75 proteins having been identified in publicly available proteomic analyses. To address this limitation, here we report new preparative methodology involving sequential solubilization of mouse SVF in guanidine hydrochloride, prior to acetone precipitation and analysis by label-free quantitative mass spectrometry. This strategy identified 126 proteins, including 83 previously undetected in SVF. Members of the seminal vesicle secretory protein family were the most abundant, accounting for 79% of all peptide spectrum matches. Functional analysis of SVF proteins identified inflammation and formation of the vaginal plug as the two most prominent biological processes. Notable additional processes included modulation of sperm function and regulation of the female reproductive tract immune environment. Together, these findings provide a robust methodological framework for future SVF studies and identify novel proteins with the potential to influence both male and female reproductive health.
Project description:We examine how NGS sequencing of sperm can provide a window as to how particular perturbations of the sperm RNA profile from baseline may be indicative of male factor infertility, and may thus provide direction as to proper course of infertility treatment for couple.
Project description:We hypothesized that seminal plasma, the acellular seminal fluid component, influences the endometrium stimulating the immune system and facilitating the implantation. We designed a randomized, double-blinded, placebo-controlled trial, and we used microarray analysis to evaluate differences in the endometrial transcriptomic profile after vaginal seminal plasma application. Differential gene pathways analysis showed an upregulation of pathways associated with the immune response, cell viability, proliferation and cellular movement, implantation, embryo development, oocyte maturation and angiogenesis. We compared our results with similar studies in pigs, mice and in vitro human endometrial cells and found similar and found comparable results. Our data show that seminal plasma has a positive effect on the endometrium during the implantation window.
Project description:We examine how NGS sequencing of sperm can provide a window as to how particular perturbations of the sperm RNA profile from baseline may be indicative of male factor infertility, and may thus provide direction as to proper course of infertility treatment for couple. NGS RNA-seq of 72 sperm samples from male partner of couples undergoing fertility treatment
Project description:Currently, the molecular mechanisms for the cause of male infertility are still poorly understood. Our previous study has demonstrated that PIWI-interacting RNAs (piRNAs) are downregulated in seminal plasma of infertile patients and can serve as molecular biomarkers for male infertility. However, the source and mechanism for the dysregulation of piRNAs remain obscure. In this study, we found that exosomes are present in high concentrations in human seminal plasma and confirmed that piRNAs are predominantly present in the exosomal fraction of seminal plasma. Moreover, we showed that piRNAs were significantly decreased in exosomes of asthenozoospermia patients compared with the fertile controls. By systematically screening piRNA profiles in sperms of fertile controls and asthenozoospermia patients, we found that piRNAs were parallelly reduced during infertility. At last, we investigated the expression of proteins that are essential for piRNAs biogenesis in sperms and therefore identified a tight correlation between the levels of spermatozoa piRNA and MitoPLD protein, suggesting that the loss-of-function of MitoPLD could cause a severe defect of piRNA accumulation in sperms. In summary, this study identified a parallel reduction of piRNAs and MitoPLD protein in sperms of infertile patients, which may provide pathophysiological clues about the development of infertility.
Project description:To examine the effect of seminal fluid on the whole genome expression profile of endometrial tissue following mating, RNA was extracted from endometrial tissue collected 8 h after CBAF1 females were mated with intact Balb/c males and compared to RNA from endometrial tissue of females mated with seminal fluid deficient SVX/VAS Balb/c males. This comparison controlled for ovarian hormone status, exposure to the male and mating activity, and the neuroendocrine response to cervical and vaginal stimulus at mating, so that changes in endometrial gene expression could be attributed specifically to contact with seminal fluid. The endometrial RNA from n=16 individual females was pooled into four independent biological replicates per treatment group (n=4 endometrial samples per replicate) and expression profiles were analyzed by Affymetrix microarray. Seminal fluid exposure induced a clear difference in the profile of genes expressed in the endometrium with a total of 335 genes were differentially regulated with a fold-change greater than 1.5 and p<0.05. Of these, 190 genes were upregulated and 145 genes were downregulated following contact with seminal fluid. Bioinformatics analysis revealed TLR4 signaling as a strongly predicted upstream regulator activated by the differentially expressed genes.Additional experiments confirmed the role of TLR4 with the absence of TLR4 in TLR4 null mice resulting in a failure for seminal fluid to induce endometrial Csf3, Cxcl2, Il6 and Tnf expression. This study provides evidence that TLR4 contributes to seminal fluid modulation of the periconception immune environment. Activation of TLR4 signaling by microbial or endogenous components of seminal fluid is thus implicated as a key element of the female tract response to seminal fluid at the outset of pregnancy in mice.