Endometriosis is a chronic inflammatory condition characterized by the presence of endometrium-like tissue outside the uterine cavity. It is estimated to affect approximately 10% of women of reproductive age and it is associated with chronic pelvic pain and infertility, among other symptoms. Endometriosis involves complex changes in the body's cells and immune response. For this reason, the goal of this observational study is to characterize the functional, molecular, and immunological alterations in menstrual blood-derived stem cells (MenSCs) and differentiated decidual stromal cells in women with endometriosis; to validate these findings in endometrial tissue and endometriomas; and to establish their correlation with clinical parameters, with the aim of identifying key pathogenic mechanisms and potential therapeutic targets. The main questions it aims to answer are: * Are there functional and molecular changes in MenSCs from patients with endometriosis compared to healthy volunteers? * Are there any variations in the immune properties of MenSCs throughout the menstrual bleeding period in patients with endometriosis compared to healthy volunteers? * Are these changes also present in endometrial tissue and endometrioma samples? * Can these changes be correlated with clinical parameters in patients with endometriosis? * Can MenSCs serve as a potential therapeutic target for endometriosis? Some participants will be asked to provide menstrual blood on a single day, while others will provide samples during the first five days of menstruation. Additionally, all participants will answer questionnaires about their diet, physical activity, stress, and pain levels. Therefore, the study does not involve the evaluation of a specific intervention on the participants. The results will enable the identification of key altered mechanisms and potential therapeutic targets, thereby contributing to the development of more effective strategies for the diagnosis and treatment of the disease.
Study Type
OBSERVATIONAL
Enrollment
40
Menstrual-blood derived stem cells will be isolated from the menstrual blood of patients with endometriosis and healthy volunteers to be characterized in vitro.
Universitat Rovira i Virgili. Facultat de Medicina i Ciències de la Salut
Reus, Tarragona, Spain
ACTIVE_NOT_RECRUITINGHospital Universitari Joan XXIII
Tarragona, Tarragona, Spain
RECRUITINGMenSCs proliferation rate
Evaluation of the expansion capacity of MenSCs from endometriosis patients vs. healthy controls using the MTT assay.
Time frame: Through study completion (average of 3 years)
Phenotypic characterization of MenSCs
Evaluation of surface marker expression using flow cytometry to determine the phenotype of MenSCs from endometriosis patients and healthy controls.
Time frame: Through study completion (average of 3 years).
MenSC chemotaxis
Assessment of the number of T-lymphocytes and monocytes capable of migrating through a transwell membrane under the stimuli of conditioned media from MenSCs obtained from endometriosis patients and healthy controls.
Time frame: Through study completion (average of 3 years)
MenSC invasion capacity
Assessment of the number of MenSCs from endometriosis patients vs. healthy controls capable of degrading a Matrigel on a transwell membrane and migrating toward a chemoattractant.
Time frame: Through study completion (average of 3 years)
Multilineage capacity of MenSCs
The differentiation potential of MenSCs from endometriosis patients vs. healthy controls towards osteocytes, chondrocytes and adipocytes will be assessed by using specific media.
Time frame: Through study completion (average of 3 years)
Decidualitzation response to progesterone of the decidual cells differentiated from MenSCs from endometriosis patients and healthy volunteers
MenSCs will be cultured in presence of 8-Br-cAMP during 14 days, and the assessment of morphology and expression of decidual markers will be performed by RT-qPCR and ELISA assay.
Time frame: Through study completion (average of 3 years).
Apoptosis resistance assessment of the decidual cells differentiated from MenSCs from endometriosis patients and healthy volunteers
Apoptosis resistance will be evaluated by Annexin V/PI flow cytometry following induction by TNFα. Additionally, the expression of key apoptotic mediators will be quantified using RT-qPCR and Western blot.
Time frame: Through study completion (average of 3 years).
Cytokine profile of the secretome of MenSCs from endometriosis patients and healthy volunteers and their differentiated decidual cells.
The conditioned media will be assessed to determine the cytokines secreted using an ELISA assay.
Time frame: Through study completion (average of 3 years).
Cytokine profile of the endometrial and endometrioma tissues
The levels of cytokines in the conditioned media from cultured explants will be assessed by ELISA assay.
Time frame: Through study completion (average of 3 years).
Cytokine profile of the secretome of immune cells populations
The levels of cytokines in the conditioned media from lymphocytes and macrophages will be assessed by ELISA assay.
Time frame: Through study completion (average of 3 years)
Cytokine profile of the peripheral blood plasma
The levels of cytokines in peripheral blood plasma from endometriosis patients and healthy volunteers will be determined by ELISA assay.
Time frame: Through study completion (average of 3 years)
Gene expression profile of MenSCs from endometriosis patients and healthy volunteers and their differentiated decidual cells
Total RNA expression of targeted genes will be analyzed by RT-qPCR.
Time frame: Through study completion (average of 3 years)
Gene expression profile of the endometrial and endometrioma tissues
Total RNA expression of targeted genes will be analyzed by RT-qPCR.
Time frame: Through study completion (average of 3 years).
Gene expression profile of the immune cells populations
Total RNA expression of targeted genes will be analyzed by RT-qPCR.
Time frame: Through study completion (average of 3 years)
Protein expression of MenSCs from endometriosis patients and healthy volunteers and their differentiated decidual cells
Analysis of the protein expression profile via Western Blot.
Time frame: Through study completion (average of 3 years)
Protein expression in endometrial and endometrioma tissues
Analysis of the protein expression profile via Western Blot.
Time frame: Through study completion (average of 3 years)
Protein expression in immune cells populations
Analysis of the protein expression profile via Western Blot.
Time frame: Through study completion (average of 3 years)
T cells activation
T cells will be exposed to conditioned media from MenSCs from healthy volunteers and endometriosis patients. Afterwards, the levels of IL-10 and IFNy will be determined by ELISA assay.
Time frame: Through study completion (average of 3 years)
T cells proliferation rate
T cells will be exposed to conditioned media from MenSCs from healthy volunteers and endometriosis patients. Afterwards, their proliferative capacity will be assessed by MTT assay.
Time frame: Through study completion (average of 3 years)
Cytokine secretion profile of macrophages
The levels of cytokines in the supernatant of polarized macrophages after incubation with MenSC-derived conditioned media from endometriosis patients and healthy volunteers will be determined by ELISA assay.
Time frame: Through study completion (average of 3 years)
Gene expression profile of macrophages
Total RNA expression of targeted genes will be analyzed by RT-qPCR.
Time frame: Through study completion (average of 3 years)
Protein expression of macrophages
Analysis of the protein expression profile via Western Blot.
Time frame: Through study completion (average of 3 years)
Phagocytic capacity of macrophages
Assessment of the macrophages' ability to engulf fluorescently labeled apoptotic bodies from MenSCs via flux cytometry and fluorescence microscopy.
Time frame: Through study completion (average of 3 years)
Angiogenic potential of macrophages conditioned media
Evaluation of the ability of macrophage-derived conditioned media to induce tube formation (Matrigel) in Human Umbilical Vein Endothelial Cells (HUVECs).
Time frame: Through study completion (average of 3 years)
Exploration of targeted interventions to reverse the pathological phenotype of MenSCs and macrophages derived from menstrual blood.
Signaling pathways and genes involved in the altered phenotype will be blocked and silenced, respectively, to evaluate if these actions reverse the cellular response to the levels of the healthy volunteers' cells.
Time frame: Through study completion (average of 3 years).
Correlation of cellular, tissue and molecular findings with patients' clinical parameters.
The associations among the functional and immunological properties of MenSCs and macrophages, clinical parameters and dietary and behavioral habits will be stablished. These clinical parameters include endometriosis stage, pain and fertility status, and the extent and localization of the endometriosis foci.
Time frame: Through study completion (average of 3 years).
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