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Analysis of lncRNA expression in endometriosis infertility, tubal disease infertility and normal subjects

ABSTRACT: Analysis of lncRNA expression in endometriosis infertility, tubal disease infertility and normal subjects

PROVIDER: PRJNA1090075 | ENA |

REPOSITORIES: ENA

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Analysis of lncRNA expression in endometriosis infertility, tubal disease infertility and normal subjects

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Exosomal microRNAs in tubal fluid may be involved in tubal epithelial structure damage and reproductive dysfunction associated with tubal endometriosis.

Project description:Endometriosis (EM) is a well-recognized disease that causes infertility in women of reproductive age. Tubal EM adversely affects fallopian tube function and female reproductive outcomes. Our study aimed to evaluate the effect of tubal EM on tubal epithelial ultrastructure and to explore the possible mechanisms of tubal infertility by analyzing the differential expression of exosomal microRNAs (miRNAs) in tubal fluid. We obtained human fallopian tube epithelium and tubal fluid samples from patients with and without tubal EM. We used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to assess ultrastructural changes. Exosomal miRNAs in tubal fluid were extracted for microarray. We visualized epithelial damage in the tubal EM group using electron microscopy. The number of organelles decreased, and organelle structure was destroyed. A total of 14 differentially expressed exosomal miRNAs were detected in tubal fluid (fold change >2 and P value <0.05). Four miRNAs (miR-1273f, miR-5699-5p, miR-6087, miR-6747-5p) were validated by quantitative real-time polymerase chain reaction (qRT-PCR). Bioinformatic analysis showed that most of the target genes participated in embryo transport, regulation of cell communication, anatomical structure morphogenesis, and immune system processes. We verified that tubal EM damaged the tubal epithelial ultrastructure in human specimens and identified differentially expressed exosomal miRNAs in tubal liquid. These findings may elucidate the pathogenesis of tubal EM-associated infertility.

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Peritoneal fluid from subjects with endometriosis regulates gene expression in cultured endometrial cells

Project description:Purpose: This study was undertaken to evaluate the effect of peritoneal fluid from subjects with differing stages of endometriosis on gene expression in endometrial stromal cells. Methods: Peritoneal fluid from subjects with minimal, moderate, and severe stages of endometriosis or without endometriosis (controls) was collected, filtered and separated from peritoneal cells. Telomerase – immortalized human endometrial stromal cells (T-HESC) were treated with the peritoneal fluid samples for 48 hours. RNA was isolated from treated cells and processed for DNA microarray. Results: 162 genes were found to be differentially expressed in T-HESC cells treated with peritoneal fluid from subjects with differing stages of endometriosis. Three of the differentially expressed genes were chosen for confirmatory studies using quantitative RT-PCR; these were interleukin 8, corin and matrix metalloproteinase 3. The primary gene ontologies identified by microarray highlight functions expected to be involved in the establishment of endometriosis, such as proteases, signal transduction, defense and immune system, cell adhesion, cell cycle and cell death, cytoskeleton, transcriptional regulation and translation, extracellular matrix, and enzymes and metabolism. Conclusions: This study demonstrated that the severity of endometriosis affected the factors present in peritoneal fluid which, in turn, affected gene expression in endometrial stromal cells.

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Project description:Endometriosis is a common disease seen by gynecologists. Clinical features involve pelvic pain and unexplained infertility. Although endometriosis is pathologically characterized by endometrial tissue outside the normal uterine location, endometriosis is otherwise not easily explained. Endometriomas, endometriotic cysts of the ovary, typically cause pain and distortion of pelvic anatomy. To begin to understand the pathogenesis of endometriomas, we carried out transcriptome:microRNAome analysis of endometriomas and eutopic endometrium, using gene expression arrays and next generation small RNA sequencing technology. Keywords: two group comparison Patients undergoing surgery for endometriomas, suspected endometriosis, pelvic pain, abnormal uterine bleeding, pelvic organ prolapse, or uterine leiomyomas were approached for participation. After informed consent, the patients underwent scheduled surgical procedure. Tissues were collected either as cyst wall of endometrioma or endometrial curettage of hysterectomy specimen and placed directly into RNALater (Ambion, Austin, TX) and eventually frozen at -80?C. Samples were designated as endometriomas or non-endometriosis control endometrium based on surgical pathology reports. Total RNA was isolated from frozen tissue. High quality RNA was subjected to Illumina’s Human WG-6 version 2.0 BeadChips (Illumina). . *** This Series represents the transcriptome component of the study. ***

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