Pathophysiology Of Endometriosis Research Articles

Endometriotic Tissue-derived Exosomes Downregulate NKG2D-mediated Cytotoxicity and Promote Apoptosis: Mechanisms for Survival of Ectopic Endometrial Tissue in Endometriosis.

Endometriosis, affecting 10% of women, is defined as implantation, survival, and growth of endometrium-like/endometriotic tissue outside the uterine cavity, causing inflammation, infertility, pain, and susceptibility to ovarian cancer. Despite extensive studies, its etiology and pathogenesis are poorly understood and largely unknown. The prevailing view is that the immune system of endometriosis patients fails to clear ectopically disseminated endometrium from retrograde menstruation. Exosomes are small extracellular vesicles that exhibit immunomodulatory properties. We studied the role of endometriotic tissue-secreted exosomes in the pathophysiology of endometriosis. Two exosome-mediated mechanisms known to impair the immune response were investigated: 1) downregulation of NKG2D-mediated cytotoxicity and 2) FasL- and TRAIL-induced apoptosis of activated immune cells. We showed that secreted endometriotic exosomes isolated from supernatants of short-term explant cultures carry the NKG2D ligands MICA/B and ULBP1-3 and the proapoptotic molecules FasL and TRAIL on their surface, i.e., signature molecules of exosome-mediated immune suppression. Acting as decoys, these exosomes downregulate the NKG2D receptor, impair NKG2D-mediated cytotoxicity, and induce apoptosis of activated PBMCs and Jurkat cells through the FasL- and TRAIL pathway. The secreted endometriotic exosomes create an immunosuppressive gradient at the ectopic site, forming a "protective shield" around the endometriotic lesions. This gradient guards the endometriotic lesions against clearance by a cytotoxic attack and creates immunologic privilege by induction of apoptosis in activated immune cells. Taken together, our results provide a plausible, exosome-based mechanistic explanation for the immune dysfunction and the compromised immune surveillance in endometriosis and contribute novel insights into the pathogenesis of this enigmatic disease.

Endometriotic Tissue-derived Exosomes Downregulate NKG2D-mediated Cytotoxicity and Promote Apoptosis: Mechanisms for Survival of Ectopic Endometrial Tissue in Endometriosis.

Endometriosis, affecting 10% of women, is defined as implantation, survival, and growth of endometrium-like/endometriotic tissue outside the uterine cavity, causing inflammation, infertility, pain, and susceptibility to ovarian cancer. Despite extensive studies, its etiology and pathogenesis are poorly understood and largely unknown. The prevailing view is that the immune system of endometriosis patients fails to clear ectopically disseminated endometrium from retrograde menstruation. Exosomes are small extracellular vesicles that exhibit immunomodulatory properties. We studied the role of endometriotic tissue-secreted exosomes in the pathophysiology of endometriosis. Two exosome-mediated mechanisms known to impair the immune response were investigated: 1) downregulation of NKG2D-mediated cytotoxicity and 2) FasL- and TRAIL-induced apoptosis of activated immune cells. We showed that secreted endometriotic exosomes isolated from supernatants of short-term explant cultures carry the NKG2D ligands MICA/B and ULBP1-3 and the proapoptotic molecules FasL and TRAIL on their surface, i.e., signature molecules of exosome-mediated immune suppression. Acting as decoys, these exosomes downregulate the NKG2D receptor, impair NKG2D-mediated cytotoxicity, and induce apoptosis of activated PBMCs and Jurkat cells through the FasL- and TRAIL pathway. The secreted endometriotic exosomes create an immunosuppressive gradient at the ectopic site, forming a "protective shield" around the endometriotic lesions. This gradient guards the endometriotic lesions against clearance by a cytotoxic attack and creates immunologic privilege by induction of apoptosis in activated immune cells. Taken together, our results provide a plausible, exosome-based mechanistic explanation for the immune dysfunction and the compromised immune surveillance in endometriosis and contribute novel insights into the pathogenesis of this enigmatic disease.

P-326 Improving quality of life and awareness of endometriosis patients using personalized web app with indications about non pharmacological support and lifestyle changes

Abstract Study question Is it possible to improve quality of life of endometriosis patients using a personalized web app suggesting non-pharmacological interventions and lifestyle changes ? Summary answer A personalized web app is useful to improve awareness and quality of life of endometriosis patients What is known already Our project is based on four main considerations: first, there is a lack of information and awareness about endometriosis symptoms and possible treatments among affected patients; second, there is no definitive cure yet and nor medications nor surgery can be considered as definitive solution; third, web instruments and social media are useful nowadays to spread knowledges about the disease; fourth, non-pharmacological support is a real instrument to better deal with the symptoms and improve quality of life. Our web app is located in this context. Study design, size, duration This is a pilot study structured on one year follow-up of symptoms and quality of life of patients with confirmed endometriosis, giving them non-pharmacological suggestions and lifestyle changes useful to better face the pathology using a personalized web app. So far, we collected data from 50 patients. Preliminary results will be based on six months followup. The project started in December 2023 Participants/materials, setting, methods Participants are patients with diagnosis of endometriosis -ultrasound,mri and surgery - between 24-45 years old, using a personalized web app in which suggestions about non-pharmacological support/lifestyle changes, given by a multidisciplinary team, are contained. Indications received are based on pelvic symptoms - dysmenorrhea, dyspareunia, chronic pelvic pain- and main systemic symptoms - chronic fatigue syndrome, migraine, bowel implications/endobelly, urinary disfunctions. Quality of life before and after the indications received was measured using EHP-30 questionnaire. Main results and the role of chance A multidisciplinary team - specialists in gynaecology, infertility, urology, pelvic floor, nutrition, chronic pain, acupuncture, psychology, yoga/pilates, law- created a web app in which indications about non-pharmacological treatments/lifestyle changes to face endometriosis symptoms are contained. We divided the indications based on main pelvic symptoms of endometriosis - dysmenorrhea, dyspareunia, chronic pelvic pain- and main systemic symptoms - chronic fatigue syndrome, migraine, bowel implications/endobelly, urinary disfunctions. For each protocol, in validation by Italian ministry of health and structured in six months indications to be followed, we gave correct informations about the pathophysiology of endometriosis, related symptoms and how to recognize them, which specialist patients should consult, what kind of supplements are useful to reduce chronic inflammation in addition/without medical therapy, implications on fertility, nutrition indications, psycological support, physical activity, when surgery should be considered or where to refer to have a proper treatment. We also created the first online community formed by specialists and patients: patients web groups are useful to share experiences, but there is risk of misinformation. The presence of specialists in a web-based community with patients could also make the difference. So far, following our indications, 50 patients reported a significant improvement in quality of life Limitations, reasons for caution Limitation in patients selection is the difference in methods of diagnosis, not always performed by specialists. We do not find reasons for caution: non-pharmacological support exists. Correct informations in unique source raise awareness, the role of supplements, pelvic floor treatments, acupuncture, nutrition, psycological support are widely recognized in literature Wider implications of the findings We aim to properly support patients in management of endometriosis. With easily wide-spreading instruments such a web app created by specialists, supported by social media and online community, we also aim to reduce the diagnostic delay. Results so far indicate improvement in quality of life among the participants. Trial registration number not applicable

O-146 Serum Progesterone levels do not differ between patients with endometriosis and unaffected patients who conceive after Hormone Replacement Therapy-Frozen Embryo Transfer (HRT-FET) cycles

Abstract Study question Do endometriosis women who achieve a live birth (LB) after Hormone Replacement Therapy-Frozen Embryo Transfer (HRT-FET) have different transfer-day progesterone levels than controls? Summary answer In women achieving a LB after HRT-FET, serum progesterone levels on the day of the transfer did not differ between endometriosis and unaffected patients. What is known already In HRT-FET, several studies have highlighted the correlation between serum progesterone levels at the time of frozen embryo transfer and LB rates. In the pathophysiology of endometriosis, progesterone resistance is typically described in the eutopic endometrium. This has led to the hypothesis that women with endometriosis may require higher progesterone levels to achieve a LB, especially in HRT-FET cycles without a corpus luteum. Study design, size, duration We conducted an observational cohort study at the university-based reproductive medicine center of our institution, focusing on women who underwent a single autologous frozen blastocyst transfer after HRT using exogenous estradiol and micronized vaginal progesterone for endometrial preparation between January 2019 and December 2021. Women were included only once during the study period. Serum progesterone levels were measured on the morning of the FET by a single laboratory. Participants/materials, setting, methods Patients were divided into groups based on whether they had endometriosis or not and whether they achieved a LB. The diagnosis of endometriosis was based on published imaging criteria (transvaginal sonography/magnetic resonance imaging) and/or confirmed histology. The primary outcome was progesterone levels on the day of the HRT-FET leading to a LB in patients with endometriosis compared to unaffected women. Subgroup analyses were performed based on the presence of deep infiltrating endometriosis or adenomyosis. Main results and the role of chance A total of 1784 patients were included. The mean age of the women was 35.1 ± 4.1 years. Five hundred and sixty women had endometriosis, while 1224 did not. 179/560 (32.0%) with endometriosis and 381/1224 (31.2%) without endometriosis achieved a LB. Among women who achieved a LB after HRT-FET, there was no significant difference in the mean progesterone level on the day of the HRT-FET between those with endometriosis and those without (13.6 ± 4.3 ng/mL versus 13.2 ± 4.4 ng/mL, respectively; p = 0.302). In the subgroup of women with deep infiltrating endometriosis (n = 142) and adenomyosis (n = 100), the mean progesterone level was 13.1 ± 4.1 ng/mL and 12.6 ± 3.7 ng/mL, respectively, with no significant difference compared to endometriosis-free patients. After adjustment for BMI, parity, duration of infertility, and tobacco use, neither the presence of endometriosis (coefficient 0.38; 95% CI -0.63 to 1.40; p = 0.457) nor the presence of adenomyosis (coefficient 0.97; 95%CI -0.24 to 2.19; p = 0.114) was associated with the progesterone level on the day of HRT-FET. Among women who did not conceive, there was no significant difference in the mean progesterone level on the day of the HRT-FET between those with endometriosis and those without (p = 0.709). Limitations, reasons for caution The primary limitation of our study is associated with its observational design. Extrapolating our results to other laboratories or different routes and/or dosages of administering progesterone also requires validation. Wider implications of the findings This study shows that patients diagnosed with endometriosis do not require higher progesterone levels on the day of a frozen blastocyst transfer to achieve a LB in hormonal replacement therapy cycles. Trial registration number NA

Features of intestinal microbiota taxonomic composition and their relation with hormonal and immune status assessed in women with external genital endometriosis

Aim: systemically assessed characteristics of intestinal microbiota taxonomic composition in relation to parameters of hormonal and immune status in patients with external genital endometriosis (EGE).Materials and Methods. The controlled cross-sectional study included 33 patients with ЕGE comprising main group, and 30 healthy women enrolled to control group. All women underwent assessment of hormonal status and cytokine expression levels in peripheral blood. Level of blood hormones estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) was carried out using enzyme-linked immunosorbent and chemiluminescent assays. Expression levels of cytokines such as interleukin (IL) IL-6, IL-8, tumor necrosis factor alpha (TNF-α) were analyzed by an enzyme-linked immunosorbent assay. Taxonomic composition of intestinal microbiota at the level of phyla and genera was carried out by 16S ribosomal RNA genesequencing. Microbial community α-diversity, the Chao1, ACE, and Sobs indices were used.Results. The concentration of blood E2 in main group was significantly higher compared to control group. Also, women with ЕGE had higher plasma concentrations for IL-6, IL-8, IL-17 and TNF-α compared to those in control group (p < 0.001). While analyzing bacterial community α-diversity in main group, Chao1 index was found to be significantly decreased. At the phylum level, the Firmicutes/Bacteroidetes ratio was increased in patients with ЕGE compared with that in control group. Among the 20 largest genera in patients with ЕGE, significant abundance was observed for Lachnospira, Blautia, Dorea, Streptococcus, Eubacterium hallii_group paralleled with significant decline in Bifidobacterium and Eubacterium eligens_group. A positive correlation was obtained between estrogen levels and the number of representatives from the genera Eubacterium hallii_group and Streptococcus, IL-8 and Streptococcus, TNF-α and Streptococcus and Lachnospira, as well as a negative correlation between TNF-α and Bifidobacterium.Conclusion. A relation between dysbiotic intestinal alterations and developing endometriosis was found. The identified correlations between altered taxonomic composition of the intestinal microflora and parameters of hormonal and immune status in patients with ЕGE suggest that intestinal microbiota is involved in pathophysiology of endometriosis.

Proangiogenic properties of complement protein C1q can contribute to endometriosis.

Endometriosis (EM) is defined as the engraftment and proliferation of functional endometrial-like tissue outside the uterine cavity, leading to a chronic inflammatory condition. While the precise etiology of EM remains elusive, recent studies have highlighted the crucial involvement of a dysregulated immune system. The complement system is one of the predominantly altered immune pathways in EM. Owing to its involvement in the process of angiogenesis, here, we have examined the possible role of the first recognition molecule of the complement classical pathway, C1q. C1q plays seminal roles in several physiological and pathological processes independent of complement activation, including tumor growth, placentation, wound healing, and angiogenesis. Gene expression analysis using the publicly available data revealed that C1q is expressed at higher levels in EM lesions compared to their healthy counterparts. Immunohistochemical analysis confirmed the presence of C1q protein, being localized around the blood vessels in the EM lesions. CD68+ macrophages are the likely producer of C1q in the EM lesions since cultured EM cells did not produce C1q in vitro. To explore the underlying reasons for increased C1q expression in EM, we focused on its established pro-angiogenic role. Employing various angiogenesis assays on primary endothelial endometriotic cells, such as migration, proliferation, and tube formation assays, we observed a robust proangiogenic effect induced by C1q on endothelial cells in the context of EM. C1q promoted angiogenesis in endothelial cells isolated from EM lesions (as well as healthy ovary that is also rich in C1q). Interestingly, endothelial cells from EM lesions seem to overexpress the receptor for the globular heads of C1q (gC1qR), a putative C1q receptor. Experiments with siRNA to silence gC1qR resulted in diminished capacity of C1q to perform its angiogenic functions, suggesting that C1q is likely to engage gC1qR in the pathophysiology of EM. gC1qR can be a potential therapeutic target in EM patients that will disrupt C1q-mediated proangiogenic activities in EM.

The Known, the Unknown and the Future of the Pathophysiology of Endometriosis.

Endometriosis is one of the most common causes of chronic pelvic pain and infertility, affecting 10% of women of reproductive age. A delay of up to 9 years is estimated between the onset of symptoms and the diagnosis of endometriosis. Endometriosis is currently defined as the presence of endometrial epithelial and stromal cells at ectopic sites; however, advances in research on endometriosis have some authors believing that endometriosis should be re-defined as "a fibrotic condition in which endometrial stroma and epithelium can be identified". There are several theories on the etiology of the disease, but the origin of endometriosis remains unclear. This review addresses the role of microRNAs (miRNAs), which are naturally occurring post-transcriptional regulatory molecules, in endometriotic lesion development, the inflammatory environment within the peritoneal cavity, including the role that cytokines play during the development of the disease, and how animal models have helped in our understanding of the pathology of this enigmatic disease.

The mysterious association between adiponectin and endometriosis.

Adiponectin is a pleiotropic cytokine predominantly derived from adipose tissue. In addition to its role in regulating energy metabolism, adiponectin may also be related to estrogen-dependent diseases, and many studies have confirmed its involvement in mediating diverse biological processes, including apoptosis, autophagy, inflammation, angiogenesis, and fibrosis, all of which are related to the pathogenesis of endometriosis. Although many researchers have reported low levels of adiponectin in patients with endometriosis and suggested that it may serve as a protective factor against the development of the disease. Therefore, the purpose of this review was to provide an up-to-date summary of the roles of adiponectin and its downstream cytokines and signaling pathways in the aforementioned biological processes. Further systematic studies on the molecular and cellular mechanisms of action of adiponectin may provide novel insights into the pathophysiology of endometriosis as well as potential therapeutic targets.

Increased Gene Expression of LITAF, TNF-α and BCL6 in Endometrial Tissues of Women with Endometriosis: A Case-Control Study.

Endometriosis, as a common inflammatory chronic disease is characterized by endometrial tissue growth outside the uterine cavity. It was reported that lipopolysaccharides (LPS) activate a transcription factor called LPSinduced tumor necrosis factor-alpha (LITAF) in macrophages, which induced transcription of cytokine genes such as tumor necrosis factor alpha (TNF-α). B-cell lymphoma 6 protein (BCL6) is a transcription factor which expression was increased in endometrial tissues of infertile women with endometriosis. In addition, it was shown that mRNA and protein of LITAF and BCL6 were inversely related in mature B lymphocytes and B-Cell lymphomas. Accordingly, we investigated gene expression of LITAF, BCL6 and ,TNF-α in eutopic and ectopic endometrial tissues of women with endometriosis compared to the controls. In this case-control study, 10 women with endometriosis (endometriosis group) and 10 women without endometriosis (control group) enrolled after diagnostic laparoscopy. Real-time polymerase chain reaction (PCR) technique was used to quantitatively analyze gene expression. One-Way ANOVA was used for data analysis. This study showed that LITAF gene expression was significantly higher in ectopic endometrial tissues compared to the control samples. Expression level of BCL6 gene was significantly increased in the ectopic tissues of women with endometriosis compared to the control and eutopic samples. Although TNF-ɑ gene expression was increased in the ectopic lesions compared to the eutopic and control endometrial samples, these differences were not significant. The results suggested that over-expression of these inflammatory genes in ectopic endometrial lesions can be considered as a molecular scenario in pathophysiology of endometriosis by induction of inflammatory cascades and cellular proliferation.

Growing evidence that endometriosis is a systemic disease

The pathophysiology of endometriosis remains unclear. Retrograde menstruation could be a phenomenon that initiates the process, but it may not explain the entire pathophysiology of endometriosis. Current evidence suggests that endometriosis is a type of chronic inflammatory disease. Many conditions that affect the vascular endothelium, including atherosclerosis, cardiovascular disease and pre-eclampsia, have been shown to be associated with endometriosis. Evidence to date suggests a complex interaction in endometriosis between angiogenesis, hormones and immunological changes stemming from chronic inflammation, with the inflammatory cells releasing cytokines and chemokines including tumour necrosis factor-α (TNF-α). Indeed, TNF-α is considered to be one of the possible markers of endometriosis in the blood, endometrium or menstrual blood. We emphasize the importance of pursuing research for novel and safer anti-inflammatory and immunomodulatory drugs that can be used by patients with endometriosis on a long-term basis.

MRNA Expression and DNA Methylation of CXCL16 in Menstrual Blood and Endometrium Tissue of Subjects with Endometriosis and Pelvic Pain

BACKGROUND: The cytokine chemokine ligand 16 (CXCL16) plays an important role in the pathophysiology of endometriosis by regulating the inflammatory response and contributing to the pain-associated endometriosis. Despite this, the impact of epigenetic modifications, such as DNA methylation, on CXCL16 has yet to be fully understood. Therefore, this research was conducted to assess both the mRNA expression and DNA methylation levels of the proinflammatory gene CXCL16 in the endometrium tissue and menstrual blood of patients with and without endometriosis.METHODS: Thirty-five women with and without endometriosis were involved in this research. Subjects' menstrual blood samples were collected using filter paper pads, meanwhile the endometrium tissue were collected by performing biopsy, from which DNA and RNA were extracted. The DNA methylation levels of the CXCL16 were measured using the pyrosequencing method following bisulfite conversion treatment. Meanwhile, the mRNA expression level was measured using the quantitative polymerase chain reaction (qPCR) method and analyzed with the Livak method.RESULTS: The mRNA expression of CXCL16 in menstrual blood of endometriosis subjects was 2.42 times higher compared to control group (p=0.030). Furthermore, the expression of CXCL16 in menstrual blood was identical to that in endometrial tissue (p=0.173). DNA methylation analysis showed that CXCL16 in the menstrual blood of endometriosis subjctes had lower methylation levels compared to controls (p=0.004), indicating hypomethylation.CONCLUSION: Increased mRNA expression and hypomethylation of CXCL16 in the menstrual blood of endometriosis patients could serve as a direct marker for diagnosing endometriosis. However, further study to validate these findings and explore the potential of CXCL16 as a diagnostic tool, and additional research involving larger patient for the cohorts study is necessary.KEYWORDS: CXCL16, DNA methylation, endometrium, menstrual blood, mRNA expression, pain

The role of peroxisome proliferator-activated receptors in endometriosis.

Endometriosis constitutes the most common cause of chronic pelvic pain in female patients and is associated with infertility. Although there is no known cause for the disease, it is a heritable condition that is determined by numerous genetic, epigenetic, and environmental aspects. Peroxisome proliferator-activated receptors (PPARs) represent nuclear receptor proteins that control gene expression. By using the MEDLINE and LIVIVO databases we conducted a literature review in order to look into the role of PPARs in the endometriosis pathophysiology and succeeded in revealing 36 pertinent publications between 2001 and 2022. In regards to PPAR expression in endometriosis, PPARγ seems to represent the most studied PPAR isoform in endometriosis and to influence various pathways involved in the disease onset and progression. It's interesting to note that diverse treatment agents targeting the PPAR system have been identified as innovative, effective therapeutic alternatives in the context of endometriosis treatment. In conclusion, PPARs appear to contribute an important role in both endometriosis pathophysiology and therapy.

The Dysregulated IL-23/TH17 Axis in Endometriosis Pathophysiology.

Endometriosis is a chronic inflammatory disease in which endometrial-like tissue grows ectopically, resulting in pelvic pain and infertility. IL-23 is a key contributor in the development and differentiation of TH17 cells, driving TH17 cells toward a pathogenic profile. In a variety of inflammatory and autoimmune disorders, TH17 cells secrete proinflammatory cytokines, including IL-17, contributing to disease pathophysiology. Our studies and others have implicated IL-17 and TH17 cell dysregulation in endometriosis, which is associated with disease severity. In this article, we address whether IL-23-driven TH17 cells contribute to cardinal features of lesion proliferation, vascularization, and inflammation in endometriosis using patient samples, representative cell lines, and our established mouse model of endometriosis. The results indicated dysregulated expression of key genes in the IL-23/TH17 axis in patient ectopic and eutopic endometrial samples and increased IL-23 protein in patient plasma compared with controls. In vitro studies using primary human TH cells determined that rIL-23 mixture treatment increased pathogenic TH17 cell frequency. Similarly, rIL-23 treatment of cell lines (12Z cells, EECCs, HUVECs, and hESCs) representative of the endometriotic lesion microenvironment increased cytokines and growth factors, which play a role in lesion establishment and maintenance. In a syngeneic mouse model of endometriosis, rIL-23 treatment altered numbers of myeloid and T cell subsets in peritoneal fluid and increased giant cells within the lesion. Lesions from rIL-23-treated mice did not reveal significant alterations in proliferation/vascularization, although trends of increased proliferation and vascularization were observed. Collectively, these findings provide insights into the impact of the IL-23/TH17 axis on local immune dysfunction and broadly on endometriosis pathophysiology.

A review: Exposure to bisphenol a analogues in non-human primates as a potential cause of endometriosis

Introduction: Bisphenol A is a synthetic compound widely used in the production of polycarbonate plastics and epoxy resins worldwide. As an environmental toxin, it has been reported in plastic equipment and utensils, water bottles and bottle tops, water supply pipes and epoxy resins that coat most of the metal food cans. It is a known endocrine-disrupting chemical and has been progressively replaced by its derivatives including bisphenol S, bisphenol F, bisphenol E, bisphenol AF, bisphenol B and tetramethyl bisphenol F. Bisphenol A and its analogues can bind to estrogen receptors and trigger multiple cellular responses at the organism level. Methods: A comprehensive literature review was done utilising electronic databases of PubMed, Google Scholar, Hinari, Connected papers and Science Direct from 1991 onwards. The articles were only included if they reported original relevant research and were limited to articles written in English. Results: Animal models, including non-human primates, have been used to study their effects on the endocrine system. Its endocrine disruption activity is reported to be the most studied effect in reproductive biology indicating that it may potentially cause endometriosis in females. Though non-human primates are closely related to humans, limited data exists on their associations between Bisphenol A exposure and its analogues and the pathophysiology of endometriosis. Conclusion: Given the current multifaceted knowledge/theory on endometriosis etiology, there is a strong necessity to conduct further biomedical research that utilises non-human primates to study the link between endocrine-disrupting chemicals and its effects on endometriosis.

Soluble MICA in endometriosis pathophysiology: Impairs NK cell degranulation and effector functions.

Endometriosis exhibits several immune dysfunctions, including deficient natural killer (NK) cell cytotoxicity. MICA (MHC class I chain-related molecule A) is induced by biological stress and soluble MICA (sMICA) negatively modulates the expression of the activating receptor, NKG2D, reducing NK cells activities. We investigated the involvement of soluble MICA in NK cell-deficient activity in endometriosis. sMICA levels (serum and peritoneal fluid-PF) were evaluated by ELISA. Circulating NK cell subsets quantification and its NKG2D receptor expression, NK cell cytotoxicity and CD107a, IFN-γ and IL-10 expressions by NK cells stimulated with K562 cells were determined by flow cytometry. We found higher sMICA levels (serum and PF) in endometriosis, especially in advanced and deep endometriosis. Endometriosis presented lower percentages of CD56dim CD16+ cytotoxic cells and impaired NK cell responses upon stimulation, resulting in lower CD107a and IFN-γ expressions, and deficient NK cell cytotoxicity. NK cell stimulation in the MICA-blocked condition (mimicking the effect of sMICA) showed decreased cytotoxicity in initial endometriosis stages and the emergence of a negative correlation between CD107a expression and sMICA levels. We suggest that soluble MICA is a potential player in endometriosis pathophysiology with involvement in disease progression and severity, contributing to NK cell impaired IFN-γ response and degranulation. NK cell compartment exhibits multiple perturbations, including quantitative deficiency and impaired cytotoxicity, contributing to inadequate elimination of ectopic endometrial tissue.

Juan-tong-yin potentially impacts endometriosis pathophysiology by enhancing autophagy of endometrial stromal cells via unfolded protein reaction-triggered endoplasmic reticulum stress

Ethnopharmacological relevanceEndometriosis (EMs) is characterized by inflammatory lesions, dysmenorrhea, infertility, and chronic pelvic pain. Single-target medications often fail to provide systemic therapeutic results owing to the complex mechanism underlying endometriosis. Although traditional Chinese medicines—such as Juan-Tong-Yin (JTY)—have shown promising results, their mechanisms of action remain largely unknown. Aim of the studyTo elucidate the therapeutic mechanism of JTY in EMs, focusing on endoplasmic reticulum (ER) stress-induced autophagy. Materials and methodsThe major components of JTY were detected using high-performance liquid chromatography–mass spectrometry (HPLC–MS). The potential mechanism of JTY in EMs treatment was predicted using network pharmacological analysis. Finally, the pathogenesis of EMs was validated in a clinical case-control study and the molecular mechanism of JTY was validated in vitro using endometrial stromal cells (ESCs). ResultsIn total, 241 compounds were analyzed and identified from JTY using UPLC–MS. Network pharmacology revealed 288 targets between the JTY components and EMs. Results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses indicated that regulating autophagy, migration, apoptosis, and inflammation were the key mechanisms of JTY in treating EMs. Meanwhile, we found that protein kinase R-like endoplasmic reticulum kinase (PERK), Beclin-1, and microtubule-associated protein light chain 3 B (LC3B) expressions were lower in endometria of patients with EMs than in those with normal eutopic endometria (p < 0.05). Additionally, during in vitro experiments, treatment with 20% JTY-containing serum significantly suppressed ESC proliferation, achieving optimal effects after 48 h. Electron microscopy revealed significantly increased autophagy flux in the JTY group compared with the control group. Moreover, JTY treatment significantly reduced the migratory and invasive abilities of ESCs and upregulated protein expression of PERK, eukaryotic initiation factor 2α (eIF2α)/phospho-eukaryotic initiation factor 2α (p-eIF2α), activating Transcription Factor-4 (ATF4), Beclin-1, and LC3BII/I, while subsequently downregulating NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and interleukin 18 (IL-18) expression. However, administration of GSK2656157—a highly selective PERK inhibitor—reversed these changes. ConclusionJTY ameliorates EMs by activating PERK associated with unfolded protein reaction, enhancing cell ER stress and autophagy, improving the inflammatory microenvironment, and decreasing the migration and invasion of ESCs.

Brain-derived neurotrophic factor (BDNF) as a potential marker of endometriosis: a systematic review and meta-analysis

BackgroundThe existing literature on the association between BDNF protein levels and endometriosis presents inconsistent findings. This systematic review and meta-analysis aim to synthesize the available evidence and evaluate the possible relationship between BDNF protein levels and endometriosis.MethodsElectronic databases (PubMed, Embase, Scopus, PsycINFO, and Web of Science) were used to conduct a comprehensive literature search from inception to June 2023. The search strategy included relevant keywords and medical subject headings (MeSH) terms related to BDNF, endometriosis, and protein levels. A random-effects model was used for the meta-analysis, and to explore heterogeneity subgroup analyses were performed. funnel plots and statistical tests were used for assessing the publication bias.ResultsA total of 12 studies were included. The pooled standardized mean difference (SMD) of BDNF levels between women with endometriosis and controls was 0.87 (95% confidence interval [CI] 0.34 to 1.39, p = 0.001; I2 = 93%). The results showed that blood levels of BDNF are significantly higher in endometriosis patients (SMD: 1.13 95% CI 0.54 to 1.73, p = 0.0002; I2 = 93%). No significant publication bias was observed based on the results of Egger’s regression test ((p = 0.15).ConclusionThis study revealed a significant difference between patients diagnosed with endometriosis and healthy control in the level of BDNF. The results indicate that women with endometriosis have higher levels of BDNF. Further studies are needed to be undertaken to investigate the role of BDNF in endometriosis pathophysiology and the diagnostic value of BDNF in endometriosis.

Unraveling the relationship between gut microbiota and site-specific endometriosis: a Mendelian randomization analysis.

Although numerous studies have illustrated the connection between gut microbiota and endometriosis, a conspicuous gap exists in research focusing on the pathogenesis of endometriosis at various sites and its linkage with infertility. In this study, we used a two-sample Mendelian randomization analysis to investigate the effect of gut microbiota on the development of endometriosis in different regions, including the uterus, ovary, fallopian tube, pelvic peritoneum, vagina, and rectovaginal septum, as well as the intestine. Additionally, we explored the correlation between gut microbiota and endometriosis-induced infertility. Genetic associations with gut microbes were obtained from genome-wide association study (GWAS) datasets provided by the MiBioGen consortium, whereas endometriosis-related GWAS data were sourced from the FinnGen dataset. In our analysis, single-nucleotide polymorphisms were used as instrumental variables, with the primary estimation of the causal effect performed via the inverse variance weighting method. Our sensitivity analyses incorporated heterogeneity tests, pleiotropy tests, and the leave-one-out method. We identified associations at the genus level between four bacterial communities and endometriosis. Subsequently, several associations between the gut microbiota and various subtypes of endometriosis at different anatomical sites were recognized. Specifically, three genera were linked with ovarian endometriosis, six genera were associated with tubal endometriosis, four genera showed links with pelvic peritoneum endometriosis, five genera were connected with vaginal and rectovaginal septum endometriosis, and seven genera demonstrated linkages with intestinal endometriosis. Additionally, one genus was associated with adenomyosis, and three genera exhibited associations with endometriosis-induced infertility. Our study elucidates associations between gut microbiota and site-specific endometriosis, thereby augmenting our understanding of the pathophysiology of endometriosis. Moreover, our findings pave the way for potential therapeutic strategies targeting gut microbiota for individuals grappling with endometriosis-related infertility.

#228 : Variability of Symptom Profiles in Individuals with Mate Kirikoopu/Endometriosis and/or Huahua Hua Kuao/Polycystic Ovary Syndrome in Aotearoa New Zealand

Background and Aims: Although opposing risk factors have been proposed for mate kirikōpū/endometriosis (oestrogen-dependence) and huahua hua kūao/polycystic ovary syndrome (PCOS; androgen dependence), there is a cohort of individuals with both endometriosis and PCOS. We propose that disease comorbidity will alter symptom profiles in comparison to those with only one disease, and that analysis of these differential profiles may hint at underlying mechanisms of disease pathology. Methods: An anonymous online survey was distributed via social media (NZ residents, &gt;18 years) and included questions related to diagnosis, symptoms, treatment and management of endometriosis and/or PCOS. Results: In total, 1323 responses (median [range] = 31 years [18–72]) were received within a two-week period (endometriosis only: n=615, 48%; PCOS only: n=459, 35%; both diseases: n=247, 19%). When asked about symptoms ‘ever experienced’, comorbid individuals report both symptoms commonly ascribed to endometriosis (e.g, dysmenorrhea) as well as those ascribed to PCOS (e.g., irregular periods) thereby exacerbating impacts on quality of life. An exception, the proportion of individuals reporting absent menstrual periods was increased in cormobid individuals (45%) relative to those with endometriosis only (21%), but decreased relative to those with PCOS only (67%), suggesting a more complex interaction between the two diseases. Conclusions: These data highlight the heterogenity of disease phenotype in individuals with endometriosis and/or PCOS. We suggest that an understanding of the intersection of these two common diseases, and a recognition of the associated phenotypic diversity, will provide avenues of research to better understand the pathophysiology of both endometriosis and PCOS.

Differential expression of interleukin-35 receptor distinguishes different subsets of granulocyte-macrophage-colony-stimulating factor-producing T helper cells in a mouse endometriosis model

The immune system contributes to the pathophysiology of endometriosis. The role of ThGM cells, which produce granulocyte macrophage-colony-stimulating factor (GM-CSF), in the pathogenesis of endometriosis remains unknown. To analyze the features of ThGM cells in endometriosis, a mouse endometriosis model was established. ThGM cells in the spleen, peritoneal fluid (PF), and endometriotic lesions (EL) were measured by flow cytometry, based on the expression of surface markers and intracellular proteins. Live ThGM cells were sorted according to chemokine receptor expression profiles and their effects on other CD4+ T cell subsets were determined by co-culture assays. An adoptive transfer assay was performed to characterize the effect of ThGM cells on endometriosis. We found that ThGM cells were present in endometriotic PF and EL. Live EL ThGM cells were enriched in CD4+CXCR3-CCR8-CCR4+CCR10+ T cells. EL ThGM cells differentially express interleukin-35 receptor (IL-35R), consisting of an IL-35R+ subset and an IL-35R- subset. The IL-35R+ subset expressed less GM-CSF, interleukin-2 (IL-2), and tumor necrosis factor-alpha (TNF-α) and proliferated slower than the IL-35R- subset. Meanwhile, the IL-35R+ subset was weaker than the IL-35R- subset in promoting the functions of Th1 and Th17 cells. ThGM cell transfer did not influence EL development but significantly alleviated pro-inflammatory cytokines in PF and ELs. Interleukin-35 (IL-35), the ligand of IL-35R, suppressed ThGM cell function and proliferation in an IL-35R-dependent manner. In summary, ThGM cells in the PF and ELs might exacerbate endometriotic inflammation. IL-35 might suppress the function of ThGM cells via IL-35R.