Project description:Progress and advancement in assisted reproductive technologies (ART) and its outcomes are limited by the importance of research of endometrial receptivity being overlooked. Due to endometrial biopsy being invasive and in vitro studies lacking reproducibility in vivo, urine is an appealing alternative biofluid source for biomarker research as it can be collected in large quantities non-invasively. The discovery of extracellular vesicles (EVs) in urine (uEVs), has also opened a new avenue in this biomarker research, with these EVs harbouring thousands of proteins that hold promise for biomarker development. In this study urine was collected from human female volunteers and samples representing the different phases of the menstrual cycle were subjected to EV isolation via differential centrifugation and size exclusion chromatography. The resulting uEVs were analysed via Nanoparticle tracking analysis (NTA) to examine the different concentration and size of particles and proteomic analysis performed using shotgun label-free mass spectrometry on the uEV samples and neat urine samples. Our results showed that uEVs were found in numbers depending on the menstrual cycle phase but uEV size was not statistically altered during different stages of the menstrual cycle. Proteomics showed 50% of proteins detected in the neat urine were also present in the uEV samples with 813 proteins were unique in the uEV samples. Proteomics analysis also showed that the menstrual cycle phase affect the uEVs proteomic profile, with some proteins shown to be significantly upregulated and downregulated during the window of implantation phase of the cycle compared to the other non-receptive periods. This data highlights that uEVs characteristics are altered depending on the menstrual cycle phase suggesting the potential of uEVs being used as biomarkers for improving fertility.In this dataset we have the neat urine results.
Project description:The objectives of the study: 1. Does the phase of the menstrual cycle alter microRNA (miRNA) plasma profiles in healthy women of reproductive age and in women with endometriosis? 2. Does this alter prospects for development of a miRNA-based diagnostic test for endometriosis?
Project description:Gene and protein expression in the breast varies relative to menopausal status (MS) and menstrual phase (MP), but cancer risk biomarker research using archived benign breast samples is usually uninformed by MS-MP data. We hypothesized that gene expression variation relative to ambient hormones can be exploited to develop markers of MS-MP that can be measured directly in breast tissue.
Project description:Despite extensive studies suggesting increased susceptibility to HIV during the secretory phase of the menstrual cycle, there is limited knowledge of the molecular mechanisms involved. We aimed to explore the ectocervical and endocervical tissue transcriptomes during the proliferative and secretory phases of the cycle using RNA sequencing (RNAseq) to identify potential signatures of susceptibility to HIV. We utilized hysterectomy tissue specimens from subjects not using hormonal contraception/treatment for gynecological conditions. Ectocervical (n=10) and endocervical tissues (n=15) were used for this study. The cycle phase was determined by assessing the histopathology of hematoxylin-and-eosin stained sections of the endometrial mucosa by gynecologic pathologists. Total RNA was isolated from tissues frozen in RNAlater (Ambion) following manufacturer’s instructions (Qiagen RNeasy Fibrous Tissue Mini Kit). After extraction, the quality and the purity of the RNA were measured by the Agilent Bioanalyzer (Agilent, Santa Clara, CA). RNA was labeled and sequenced at the RU Genomics center by using Illumina TruSeq technology (75bp, >30M coverage). The data were analyzed using Ingenuity Pathway Analysis (IPA) software (QIAGEN Inc., https://www.qiagenbioinformatics.com/products/ingenuitypathway-analysis). To inquire into changes in gene expression irrespectively of p-value cut off, pre-ranked genes based on proliferative vs. secretory phase expression were subjected to Gene Set Enrichment Analysis (GSEA) against the Hallmark Gene sets (H) and Immunologic Signatures Gene sets (collection C7) from the Molecular Signatures Database (MSigDB)(http://software.broadinstitute.org/gsea/msigdb/index.jsp). Our data show menstrual cycle phase-associated changes in the transcriptomic landscape of the endocervix and ectocervix, some of which may contribute to changes in HIV susceptibility.