Profile of circulating and exosomal miRNAs in biological fluids of infertile women with endometriosis.

• Grant number: 24/06420-6
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2025
• End date: March 31, 2027
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Thérèse Rachell Theodoro
• Grantee: Thérèse Rachell Theodoro
• Host Institution: Centro Universitário FMABC (FMABC). Santo André , SP, Brazil
• Associated researchers: Bianca Alves Vieira Bianco ; Caio Parente Barbosa ; Erik Montagna

Abstract

Endometriosis (EDT) is a chronic inflammatory gynecological disease characterized by the presence of endometrial tissue outside the uterus and is associated with reproductive complications, including infertility that affects 10%-15% of women of reproductive age. The pathophysiological mechanisms that connect endometriosis to infertility are not yet completely understood, which limits the development of effective diagnostic and therapeutic strategies. MiRNAs participate in the regulation of post-transcriptional gene expression and can influence the development and progression of endometriosis. Furthermore, exosomal miRNAs may be capable of carrying information between cells, promoting cell-cell communication and immunoregulatory functions, through exo-miRNAs, to recipient cells. Inappropriate release of exo-miRNAs can cause significant changes in biological pathways that affect disease development, supporting the concept that exosomes can serve as targeted therapies for diseases, favoring transmission of various inflammatory signals, including in endometriosis. Circulating and exosomal miRNAs can be detected in circulating body fluids, making them low-invasive biomarkers. Among the miRNAs already studied in body fluids in endometriosis, the following were selected: mir-30e-5p and mir-15b-5p, as normalizers because these miRNAs have been identified in cohort studies as normalizers through the average of mir-30e-5p and mir-15b-5p; mir-let-7b was found to have lower expression in women with endometriosis vs. without endometriosis in body fluids such as serum, in addition to being involved in the deregulation of the p53 pathway and control of the cell cycle; mir-30c-5p is shown to have increased levels in women with endometriosis versus women without endometriosis, in addition to its decrease being related to the increase in mutations in the p53 pathway as they inhibit cell proliferation, migration and invasion. The low expression of exosomal mir-30c-5p in ectopic endometrial epithelial cells of women with endometrioma, led to the overexpression of BCL9 (B9 cell lymphoma), which is a co-activator of the Wnt/²-catenin pathway, and may thus have the capacity invasive and migratory potential of endometriotic lesions; mir-214-3p is related to the angiogenesis process and increased VEGF levels in fibrotic endometriotic lesions. Patients with uterine fibroids showed increased miR-214 expression and the miR-214-PI3K-AKT pathway was suggested as one of the mechanisms to treat uterine fibroids as an abnormal disease of female blood circulation. Thus, circulating miR-214-3p may be a promising therapeutic target for fibrotic diseases, including EDT. In a rodent model for endometriosis, reduced fibrosis was observed in ectopic lesions when treated with exosomes derived from human ectopic endometrial stromal cells transfected with exo-miR-214-3p mimics. Furthermore, studies have shown downregulation of exo-miR-214-3p in ectopic lesions and eutopic endometrium of patients with endometriosis compared with eutopic endometrium of controls and lower serum level of exo-miR-214-3p in patients with EDT. The present study aims to detect a profile of circulating and exosomal miRNAs in body fluids such as peripheral blood, serum, plasma and urine, as potential biomarkers for diagnosis and/or therapeutic targets for EDT. Specific techniques will be used to collect body fluids, isolate exosomes, extract circulating miRNAs (magnetic beads) and exosomes isolated from body fluids. Furthermore, the TaqMan real-time PCR technique will be applied to amplify the specific miRNAs selected for EDT. The results of this study could lead to the development of more accurate and less invasive early diagnostic tests and more effective exosome treatment for endometriosis. (AU)