New Perspectives in Adenomyosis Pathogenesis With Epigenetic Analysis and miRNAs

Overview

Objectives: To identify differentially expressed miRNAs in the blood of adenomyosis patients in view to develop new diagnostic methods Hypotheses Circulating miRNAs may be abnormally expressed in patients suffering from adenomyosis and could be used to diagnose the disease Study Design Blood samples will be collected from healthy subjects and adenomyosis patients and miRNAs will be isolated and analyzed to detect potential differences.

The present study is based on previous research pointing out that circulating miRNAs are abnormally expressed in several pathologies including endometriosis, and can be used as a handy diagnostic tool in the clinical setting. The investigators hypothesize that miRNAs are differentially expressed in adenomyosis as well and could serve as noninvasive biomarkers to diagnose the condition. To test this hypothesis, the investigators plan to conduct a pilot study on 50 patients (25 adenomyosis patients and 25 healthy subjects), in order to compare expression of different miRNAs between the groups and determine a suitable diagnostic panel. There is approximatively 25 women undergoing hysterectomy for adenomyosis over the course of a year at the CUSL. It is also a necessary population in order to obtain statistically analyzable datas. This panel may be tested in the future in a larger population for validation purposes. Blood samples (10 ml) will be collected the day of the surgery by the anesthetist from female patients visiting the CUSL for the purpose of hysterectomy. Patients will be subdivided into two groups, namely the study group (n=25) including patients diagnosed with adenomyosis by MRI and/or ultrasonography prior surgery and the control group (n=25) consisting of patients with pathologies unrelated to the endometrium. After samples registration in CUSL biobank, they will be transferred to the research laboratory and serum will be collected immediately by centrifuging at 2500 rpm for 15 min at 4° C followed by storage at -80°C until further use. To isolate circulating miRNAs, a commercial kit (Qiagen miRNeasy serum/plasma kit) will be used and cDNA will be then synthesized using appropriate kit (Qiagen miScript II RT). Commercial miRNA arrays will be used to simultaneously quantify expression of around 1000 miRNAs and compare their expression profiles between adenomyosis patients and unaffected subjects. Individual RT-qPCR reactions will be then conducted to validate the results. Differentially expressed microRNAs in adenomyosis will be further analyzed to determine their target genes and subsequent affected biological functions, using appropriate databases (TargetScan and miRTarBase). This experimental approach will allow us to identify abnormally expressed miRNAs in adenomyosis compared to disease-free women. These can then be used as noninvasive biomarkers of the pathology and/or targeted for development of new therapeutic options.

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