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Functional microRNA involved in endometriosis.

ABSTRACT: 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 describe the first transcriptome-microRNAome analysis of endometriomas and eutopic endometrium using next-generation sequencing technology. Using this approach, we generated a total of more than 54 million independent small RNA reads from our 19 clinical samples. At the microRNA level, we found 10 microRNA that were up-regulated (miR-202, 193a-3p, 29c, 708, 509-3-5p, 574-3p, 193a-5p, 485-3p, 100, and 720) and 12 microRNA that were down-regulated (miR-504, 141, 429, 203, 10a, 200b, 873, 200c, 200a, 449b, 375, and 34c-5p) in endometriomas compared with endometrium. Using in silico prediction algorithms, we correlated these microRNA with their corresponding differentially expressed mRNA targets. To validate the functional roles of microRNA, we manipulated levels of miR-29c in an in vitro system of primary cultures of human endometrial stromal fibroblasts. Extracellular matrix genes that were potential targets of miR-29c in silico were significantly down-regulated using this biological in vitro system. In vitro functional studies using luciferase reporter constructs further confirmed that miR-29c directly affects specific extracellular matrix genes that are dysregulated in endometriomas. Thus, miR-29c and other abnormally regulated microRNA appear to play important roles in the pathophysiology of uterine function and dysfunction.

SUBMITTER: Hawkins SM

PROVIDER: S-EPMC3082329 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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Functional microRNA involved in endometriosis.

Hawkins Shannon M SM Creighton Chad J CJ Han Derek Y DY Zariff Azam A Anderson Matthew L ML Gunaratne Preethi H PH Matzuk Martin M MM

Molecular endocrinology (Baltimore, Md.) 20110324 5

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 describe the first transcriptome-microRNAome ana ...[more]

PMID: 21436257

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Project description:Are microRNAs (miRs) altered in the eutopic endometrium (EuE) of baboons following the induction of endometriosis?Induction of endometriosis causes significant changes in the expression of eight miRs, including miR-451, in the baboon endometrium as early as 3 months following induction of the disease.Endometriosis is one of the most common gynecological disorders and causes chronic pelvic pain and infertility in women of reproductive age. Altered expression of miRs has been reported in women and has been suggested to play an important role in the pathophysiology of several gynecological disorders including endometriosis.EuE was obtained from the same group of baboons before and 3 months after the induction of endometriosis. The altered expression of miR-451 was validated in the eutopic and ectopic endometrium of additional baboons between 3 and 15 months following disease induction. Timed endometrial biopsies from women with and without endometriosis were also used to validate the expression of miR-451.Total RNA was extracted from EuE samples before and after the induction of endometriosis, and miRNA expression was analyzed using a 8 × 15 K miR microarray. Microarray signal data were preprocessed by AgiMiRna software, and an empirical Bayes model was used to estimate the changes. The present study focused on quantitative RT-PCR validation of the microarray data, specifically on miR-451 and its target genes in both baboons (n = 3) and women [control (n = 7) and endometriosis (n = 19)]. Descriptive and correlative analysis of miR-451 and target gene expression was conducted using in situ hybridization and immunohistochemistry, while functional analysis utilized an in vitro 3' untranslated region (UTR) luciferase assay and overexpression of miR-451 in human endometrial and endometriotic cell lines.Induction of endometriosis results in the altered expression of miR-451, -141, -29c, -21, -424, -19b, -200a and -181a in the baboon endometrium. In the baboon, induction of endometriosis significantly decreased the expression of miR-451 at 3 months (P < 0.001), which was also associated with increased expression of its target gene YWHAZ (14.3.3?). A similar significant (P < 0.0001) decrease in miR-451 expression was observed in women with endometriosis. The 3' UTR luciferase assay confirmed the regulation of YWHAZ expression by miR-451. Furthermore, overexpression of miR-451 in 12Z cells (immortalized human endometriotic epithelial cell line) led to the decreased expression of its target YWHAZ and this was correlated with decreased cell proliferation.The study focused only on miR-451 and one of its targets, namely YWHAZ. A single miR could target number of genes and a single gene could also be regulated by number of miRs; hence, it is possible that other miRs and their regulated genes may contribute to the pathophysiology of endometriosis.Our data suggest that the presence of ectopic lesions in baboon causes changes in EuE miR expression as early as 3 months postinduction of the disease, and some of these changes may persist throughout the course of the disease. We propose that the marked down-regulation of miR-451 in both baboons and women with endometriosis increases the expression of multiple target genes. Increased expression of one of the target genes, YWHAZ, increases proliferation, likely contributing to the pathophysiology of the disease.

Project description:Endometriosis is a common, chronic gynaecological disease characterised by pelvic pain and sub-fertility. It is a complex genetic disease affecting of 6-10% women of reproductive age and up to 50% of women with infertility. Evidence from genomewide association (GWA) imputed and meta-analysis studies in women with and without endometriosis identified association on chromosome 12q22, located close to the putative candidate gene VEZT. Fine-mapping this region in our 1,029 Australian cases and 958 controls confirmed strong evidence of association for non-coding variants in the 12q22 GWA region with the best imputed SNP rs4762347 in the 3’UTR of VEZT (P=0.036; 95% CI, 1.01-1.38; OR =1.18) and four top imputed SNPs in high linkage disequilibrium (LD) with rs4762347. Bioinformatic analysis for potential functional roles for these SNPs using ENCODE data show rs4762347 is highly conserved and alters a regulatory motif for the transcription factor Nkx3. We conducted an initial experiment to evaluate gene expression of VEZT in 36 samples of endometrial tissue from endometriosis cases and controls using high-throughput gene expression RT-qPCR on a microfluidic dynamic array IFC 48x48 chip (Fluidigm Biomark). The results showed that VEZT was expressed in endometrial samples from cases and controls. There was a trend for increased expression levels of VEZT mRNAs in endometriosis cases compared to controls as well as during the secretory phase compared to proliferative phase of menstrual cycle, although these differences were not statistically significant. The top imputed SNP rs4762347 was chosen to test for its allelic effects on VEZT expression. There was suggestive evidence for differences in VEZT mRNA expression observed in allelic groups of rs4762347. VEZT mRNA expression decreased for transcripts in carriers of minor C allele (CC and CT) of rs4762347 compared to non carriers (TT) (P<0.01), but the differences were no longer significant after corrections for multiple testing, hence further gene expression studies should be conducted in a larger sample size.

Dataset Information

Functional microRNA involved in endometriosis.

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Functional microRNA involved in endometriosis.

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