Endometriotic Disease Research Articles

The Role of Peritoneal Immunity in Peritoneal Endometriosis and Related Infertility.

Endometriosis is defined as a disorder in which the glands and stroma of the endometrium grow and shed periodically outside the uterine cavity. Highly prevalent in women of reproductive age, the most common clinical manifestations are chronic pelvic pain and infertility. The pathogenesis of endometriosis may be multifactorial, including factors of anatomy, immunity, inflammation, hormones (estrogen), oxidative stress, genetics, epigenetics, and environment. There are generally three types of endometriotic disease, namely peritoneal, ovarian, and deep infiltration. For the same patient, there may be a single or multiple types concurrently. The different manifestations of these types suggests that they each have their own etiology. Numerous studies have shown that the evasion of endometrial cells from peritoneal immune surveillance helps establish and maintain peritoneal endometriosis, but the specific mechanism is not well understood. Likewise, the molecular mechanisms of endometriosis-related infertility have not been clearly elucidated. This review attempts to identify the role of peritoneal immunity in peritoneal endometriosis and related infertility, especially in the aspects of molecular mechanisms.

The appendix in endometriosis.

In the most severe stage of endometriosis, Stage IV, intestinal involvement is common. The true prevalence of endometriotic disease of the appendix in this population is not well described. A macroscopically normal looking appendix may harbour endometriosis. Our study aims to assess the role of routinely performing appendicectomy in Stage IV endometriosis surgery, and the histopathological prevalence of true appendiceal endometriosis in this population. This is a retrospective study of women undergoing surgery for Stage IV endometriosis between 2018 to 2022 in a tertiary public hospital in New South Wales, Australia. Patient demographics, age and post-operative complications were retrospectively retrieved from hospital medical records. Inclusion criteria were women with Stage IV endometriosis who underwent routine appendicectomy as part of their endometriosis surgery. Exclusion criteria were women who did not have Stage IV endometriosis, those who had cancer surgery or emergency surgery for endometriosis. The primary outcome of this study was to determine the incidence of appendiceal endometriosis. Secondary outcomes included post-operative complications and length of stay. Sixty-seven patients were included. The mean age was 36 years. All patients also underwent bowel resection for colorectal endometriosis. There were 35.8% who had confirmed appendiceal endometriosis on histopathology. Post-operative complications included port site infections, colitis, urinary tract infection and ureteric injury. There were no complications related to appendicectomy. Mean length of stay was 4.4 days. Laparoscopic appendicectomy can be safely performed at time of laparoscopic surgical excision of Stage IV endometriosis and should be routinely considered in a subset of Stage IV endometriosis patients with colorectal involvement undergoing surgery.

Stomatin-like Protein-2 Promotes Aggregation, Colonization and Migration of Endometriotic Cells.

Endometriosis is a chronic gynecological disease in women of childbearing age, which leads to infertility with risk of endometrial and ovarian cancer. The pathogenesis of endometriosis is poorly understood, and cure/treatment for it is not available, except for symptomatic treatment. The recurrence rate of endometriosis is high. SLP-2 is an inner mitochondrial membrane protein whose participation has been explained in cases of endometrial stromal cell growth, differentiation and migration, but its role in endometriosis is yet to be understood. Previous studies have found altered expression of stomatin-like protein 2 (SLP-2) in the serum of endometriotic patients. Therefore, we have studied the possible role of SLP-2 in the development of endometriosis. We found the ubiquitous and high expression of SLP-2 in the endometriotic tissue of both human endometriosis patientsand rat endometriosis model. SLP-2 is seen in the glandular epithelial cells and stromal cells in the eutopic/normal or non-endometriosis group endometrium from human subjects. Finding high expression levels of SLP-2 in endometriotic tissue and ovarian cystic cells derived from endometriosis patients, we explored the possible role of SLP-2 in the cell aggregation, colonization, migration, and invasion in the human endometriotic cells associated with the progression of the endometriosis. Transient silencing of SLP-2 by its siRNA hinders endometriotic cells, aggregation, migration, and invasion into the extracellular matrix, which confirms SLP-2 involvement in endometriotic disease onset and progression. This study unravels the ubiquitous expression of SLP-2 in the human ectopic endometrial tissue and its role in the endometriotic cell migration, colonization, aggregation, and invasion leading to endometriosis progression.

Endometriosis: A Retrospective Analysis on Diagnostic Data in a Cohort of 4,401 Patients

Endometriosis is a gynecological estrogen-dependent inflammatory disease due to ectopic endometrial tissue and often associated with pelvic pain. Despite its high prevalence, there are still uncertainties about its pathogenesis, diagnosis, and therapy.This study presents a retrospective study conducted on 4,401 endometriosis patients, 584 of which underwent laparoscopic procedures. The archived data about clinical signs, magnetic resonance imaging (MRI) results, topography of the endometriosis lesions (obtained via laparoscopy) associated diseases, sample analysis and histological findings were analyzed. Next, the statistical associations between the information for each case, provided by these diagnostic tools were determined.MRI is the most sensitive and specific diagnostic system for ovarian lesions, but poor in sensitivity and specificity for deep endometriosis lesions and not indicated for peritoneal lesions which remain the exclusive prerogative of laparoscopy. Clinical signs are essential for diagnosing deep lesions. The Ca125 and Ca19.9 markers have a poor reliability and their negativity in symptomatic patients has no clinical value, while in positive cases it could probably be used as a monitoring parameter.The results generated will help provide an accurate picture of the topography and distribution of endometriotic lesions. Correlation analyses between the data generated by the clinical-instrumental examinations and those on the site of the disease identified by laparoscopy, allow to define the predictive value of the clinical-instrumental signs in the diagnosis and localization of endometriotic disease.

Can New ENZIAN Score 2020 Represent a Staging System Improving MRI Structured Report?

Structured reporting systems for endometriotic disease are gaining a central role in diagnostic imaging: our aim is to evaluate applicability and the feasibility of the recent ENZIAN score (2020) assessed by MRI. A total of 60 patients with suspected tubo–ovarian/deep endometriosis were retrospectively included in our study according to the following criteria: availability of MR examination; histopathological results from laparoscopic or surgical treatment; patients were not assuming estro-progestin or progestin therapy. Three different readers (radiologists with 2-, 5-, and 20-years of experience in pelvic imaging) have separately assigned a score according to the ENZIAN score (revised 2020) for all lesions detected by magnetic resonance imaging (MRI). Our study showed a high interobserver agreement and feasibility of the recent ENZIAN score applied to MRI; on the other hand, our experience highlighted some limitations mainly due to MRI’s inability to assess tubal patency and mobility, as required by the recent score (2020). In view of the limitations which arose from our study, we propose a modified MRI-ENZIAN score that provides a complete structured reporting system, more suitable for MRI. The high interobserver agreement of the recent ENZIAN score applied to MRI confirms its validity as a complete staging system for endometriosis, offering a shared language between radiologists and surgeons.

A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice.

Endometriosis is a leading cause of pelvic pain and infertility. It is defined by the presence of endometrial tissue in extrauterine locations. The development of novel therapies and diagnostic tools for endometriosis has been limited due in part to challenges in studying the disease. Outside of primates, few mammals menstruate, and none develop spontaneous endometriosis. Rodent models are popular but require artificial induction of endometriosis, with many utilizing either immunocompromised mice or surgically induced disease. Recently, more attention has been given to models involving intraperitoneal injection. We present a murine model of endometriosis that integrates several features of existing endometriosis models into a novel, simplified system that relies on microscopic quantification in lieu of subjective grading. In this model, we perform hormonal stimulation of donor mice, intraperitoneal injection, systematic abdominal survey and tissue harvest, and histologic quantification that can be performed and verified at any time after necropsy. This model requires minimal resources and training; does not require expertise by lab technicians in murine survival surgery or in the identification of gross endometriotic lesions; can be used in immunocompromised, immunocompetent, and/or mutant mice; and reliably creates endometriotic lesions that are histologically consistent with human endometriotic disease.

Menstrual characteristics, bleeding and other non-pelvic-pain symptoms in women presenting with severe endometriotic disease

Introduction: The purpose of this study was to investigate the detailed nature of menstrual characteristics, bleeding and other symptoms in women with and without endometriosis. Pelvic pain symptoms in this cohort have been reported elsewhere. Method: This is a self-administered cross-sectional study and was conducted using a structural questionnaire. A total of 737 women completed the study: women with endometriosis (n = 529) and with no current gynaecological complaint, ‘control’ (n = 208). Results: Some of the key findings in this study included significant differences between the control and endometriosis subjects in terms of bleeding symptoms, increased length and heaviness of menses, rectal and bladder bleeding, infertility and obstetric complications which were all more common in the endometriosis group. Other symptoms found to have a highly significant positive association with endometriosis included abdominal bloating, low resistance to infection, fatigue, increased urinary frequency, diarrhoea and constipation (p < 0.000). Data were cross-tabulated to examine the association of a number of symptoms to each other. The relationship of symptoms to a range of psychosocial factors was investigated to determine how the effects of endometriosis impacted on the life of subjects. Most women reported that endometriosis sometimes or often interfered with daily aspects of their life. Conclusion: The range of non-pain symptoms in women with endometriosis was generally much more diverse and extensive than recognised by most clinicians.

Letrozole and the Traditional Chinese Medicine, Shaofu Zhuyu Decoction, Reduce Endometriotic Disease Progression in Rats: A Potential Role for Gut Microbiota.

We previously showed that the Chinese herbal medicine, Shaofu Zhuyu decoction (SFZYD), shrank the size of endometriotic lesions in rats with endometriosis. We therefore conducted the present study to investigate the effects of letrozole and SFZYD on gut microbiota in endometriotic rats. Rats were divided into four groups: a blank group, model group, letrozole group, and SFZY group. Ectopic lesion size and COX-2 expression in the endometrium and endometriotic lesions were compared, and the community of gut microbiota was detected using 16S rRNA gene sequencing. Both letrozole and SFZYD reduced the size of ectopic lesions as well as lowered the expression of COX-2, thus reducing the inflammatory response. Compared with the blank group, the α-diversity of gut microbiota in endometriotic rats decreased, the Firmicutes/Bacteroidetes ratio increased, and the abundance of Ruminococcaceae was reduced. The α-diversity of gut microbiota in the letrozole group was similar to that in the model group, but the Firmicutes/Bacteroidetes ratio was diminished. The α-diversity in the SFZY group was similar to that in the blank group, the Firmicutes/Bacteroidetes ratio was attenuated, and the abundance of Ruminococcaceae was elevated compared with the model group. These results indicated that the therapeutic mechanisms of both letrozole and SFZYD were related to the restoration of gut microbiota.

SAT-197 Functional Cross-Talk Between MCP-1 and Integrin Linked Kinase Is Required for Endometriosis Disease Progression

Development of painful invasive endometriotic lesions is usually a combined function of the chronic inflammation and active sequential cellular events such as migration, cell adhesion, proliferation, colonization, and invasion in endometriosis. An elevated level of monocyte chemotactic protein-1 (MCP-1) in serum and peritoneal fluid has been reported in endometriosis. Further, Integrin-Linked Kinase (ILK) communicates with extracellular signals and regulates the sequential cellular events in uterine and ovarian cancer. The functional cross-talk of MCP-1 with ILK in the immune cells leads to the high MCP-1 production and increased ILK activity. However, the role of the elevated MCP-1 level and its functional interaction with ILK in the endometriosis development is not explored yet. In order to investigate this, we used the surgically induced heterologous mouse model of endometriosis and externally treated with MCP-1 intraperitoneally for two weeks. Our results demonstrated that MCP-1 boosted the growth of endometriotic lesions mediated through ILK, FAK, VAV and RAC1 signaling axis in endometriosis. In addition, in vivo pharmacological inhibition of ILK by CPD22 restricted the growth of endometriotic lesions in mice. Flow cytometric and inflammatory cytokines profiling in the peritoneal fluid demonstrated that ILK was involved in the MCP-1 mediated macrophageF4/80+ recruitment and inflammatory response in order to support the growth of endometriotic lesions. Inhibition of ILK induced the anti-inflammatory cytokine IL-10 and concomitantly suppressed the pro-inflammatory cytokines MCP-1, TNF-α and IL-10 levels in peritoneal fluid. In vitro studies on human ovarian cystic endometriosis cells (Hs832(C).T) and normal endometrial stromal cells (T-HESCs) revealed that MCP-1 accelerated the adhesion (on collagen and fibronectin substratum), colonization, migration and 3D-spheroids formation and invasion. As the basal level of CCR2, an MCP-1 receptor, expression was higher in ovarian endometriotic cells than the normal endometrial stromal cells giving the hint for the role of MCP-1 in endometriosis. Interestingly, the siRNA mediated knockdown of ILK renders the MCP-1 induced nuclear translocation of phosphorylated ILK and also reduced these accelerated cellular events. Taken together, our results provide compelling evidence that the functional cross-talk between MCP-1 and ILK drives endometriotic disease progression.

Nerve fibre infiltration and expression in peritoneal lesions of endometriosis in a nonhuman primate model of endometriosis

Baboon ( Papio anubis) models of endometriosis are thought to mimic the early stages of spontaneous human peritoneal endometriotic disease. The objective of this study was to investigate the presence and course of nerve fibre ingrowth during peritoneal lesion formation in specimens collected at 3 months (early stage of lesion development) and 15 months (late stage of lesion development) after disease initiation compared to pelvic peritoneum (control). Five-micron sections of paraffin-embedded peritoneal lesions were obtained from normally cycling baboons with 3-month (n = 12), 15-month (n = 12) induced endometriosis and pelvic peritoneum (n = 10) from baboons with no endometriosis. Immunohistochemical staining was performed with specific antibodies: protein gene product 9.5 – broad marker of nerve fibres and neurones, neuropeptide Y – sympathetic neurones, substance P – sensory neurones, vasoactive intestinal peptide – parasympathetic neurones, nerve growth factor – development of new neurones and high-affinity receptor for nerve growth factor (tropomyosin receptor kinase A) – neuronal differential. Significantly, more nerve fibres were identified in peritoneal endometriotic lesions collected 15 months after the initiation of experimental protocols compared with 3-month and control samples (p < 0.001). Nerve fibres were immunoreactive for all the tested markers – protein gene product 9.5, neuropeptide Y, substance P, vasoactive intestinal peptide, nerve growth factor and tropomyosin receptor kinase A – indicating the presence of different types of nerve fibres. In conclusion, peritoneal lesions of endometriosis in a nonhuman primate model of endometriosis were found to be progressively and spontaneously innervated by both myelinated and unmyelinated sensory nerve fibres, parasympathetic and sympathetic neurones. These nerve fibres may play an important role in the mechanisms of pain generation in this condition.

Репродуктивне здоров’я і дисфункція щитоподібної залози

Diseases of thyroid gland, according to world statistics, are found in almost 30% of the world’s population. Thyroid dysfunctions, according to many epidemiological studies, are quite widespread in the population. Normal development of the reproductive system occurs under the influence of gonadotropic hormones of the pituitary gland and during normal functioning of the thyroid gland. In turn, the state of the reproductive system has a pronounced effect on the function of the thyroid gland, as evidenced by changes in its function during pregnancy and lactation, in patients with abnormal uterine bleeding, in girls during puberty and women during the menstrual cycle. The objective: is to evaluate the prevalence and effectiveness of diagnosis of dysfunction of thyroid dysfunction in women with reproductive health disorders to improve treatment and rehabilitation activities and quality of life. Materials and methods. In the first stage of a prospective study, 989 women of reproductive age were put through the method of continuous sampling. Based on the results of verifying the gynecological pathology, 350 patients who gave informed consent for participation in the second stage of the study, were divided into clinical groups for further comparative analysis of the prevalence of dysfunction of thyroid gland: I group – 159 (51.3%) patients with combined non-hormonal non-inflammatory pathology of reproductive organs and dyshormonal pathology of the mammary glands (genital endometriosis, adenomyosis / uterine leiomyoma in combination with endometrial hyperplasia), II group – 31 (8.9%) female with PCOS, III – 53 (15.1%) with endometriotic disease, IV group – 57 (16.3%) women with uterine leiomyoma. Absence of gynecological pathology and pathology of mammary glands at the time of the survey was found in 50 (14.3%) of women, which amounted to V group. Results. Structural and functional changes of the thyroid gland were found in 53.4% of women, while the frequency of detection of diffuse goiter I–II st. did not differ between clinical groups (p>0.05). The prevalence of autoimmune thyroiditis was higher in patients with dyshormonal pathology of the reproductive system than in healthy women (p=0.0001). The euthyroid state with combined dyshormonal and proliferative pathology occurred 1.5 to 1.9 times less frequently than in patients of other groups (p≤0.0005). Subclinical hypothyroidism was diagnosed in 39,6% of the I group of women at 12.9%, 15.1% and 14.0% in the II, III and IV groups, respectively. At the same time, the incidence of subclinical hypothyroidism was 60.6% for the I group in women who had diffuse goiter and / or autoimmune thyroiditis, with 8.6% of the manifest hypothyroidism that was absent in women with PCOS, and the prevalence of endometriosis disease and uterine leiomyomies were 1.9% and 3.5%, respectively. Conclusion. Thus, the results of a two-stage study of thyroid gland thyroid dysfunction in women with dyshormonal pathology of reproductive organs of non-inflammatory genesis confirm that dysfunction of the thyroid gland, especially hypothyroidism, is a serious problem for reproductive health, requiring changes in diagnostic and therapeutic approaches. Key words: thyroid gland, autoimmune thyroiditis, hypothyroidism, reproductive health, non-inflammatory dyshormonal diseases of reproductive organs.

Variability of genome-wide DNA methylation and mRNA expression profiles in reproductive and endocrine disease related tissues

ABSTRACTGenome-wide association studies in the fields of reproductive medicine and endocrinology are yielding robust genetic variants associated with disease. Integrated genomic, transcriptomic, and epigenomic molecular profiling studies are common methodologies used to understand the biologic pathways perturbed by these variants. However, molecular profiling resources do not include the tissue most relevant to many female reproductive traits, the endometrium, while the parameters influencing variability of results from its molecular profiling are unclear. We investigated the sources of DNA methylation and RNA expression profile variability in endometrium (n = 135), endometriotic disease tissue (endometriosis), and subcutaneous abdominal fat samples from 24 women, quantifying between-individual, within-tissue (cellular heterogeneity), and technical variation. DNA samples (n = 96) were analyzed using Illumina HumanMethlylation450 BeadChip arrays; RNA samples (n = 39) were analyzed using H12-expression arrays. Variance-component analyses showed that, for the top 10–50% variable DNA methylation/RNA expression sites, between-individual variation far exceeded within-tissue and technical variation. Menstrual-phase accounted for most variability in methylation/expression patterns in endometrium (Pm = 7.8 × 10−3, Pe = 8.4 × 10−5) but not in fat and endometriotic tissue; age was significantly associated with DNA methylation profile of endometrium (Pm = 9 × 10−5) and endometriotic disease tissue (Pm = 2.4 × 10−5); and smoking was significantly associated with DNA methylation in adipose tissue (Pm = 1.8 × 10−3). Hierarchical cluster analysis showed significantly different methylation signatures between endometrium and endometriotic tissue enriched for WNT signaling, angiogenesis, cadherin signaling, and gonadotropin-releasing-hormone-receptor pathways. Differential DNA methylation/expression analyses suggested detection of a limited number of sites with large fold changes (FC > 4), but power calculations accounting for different sources of variability showed that for robust detection >500 tissue samples are required. These results enable appropriate study design for large-scale expression and methylation tissue-based profiling relevant to many reproductive and endocrine traits.

Seminal Plasma Promotes Lesion Development in a Xenograft Model of Endometriosis

The factors that predispose one-tenth of reproductive-aged women to endometriosis are poorly understood. We determined that genetic deficiency in transforming growth factor β1 impairs endometriosis-like lesion growth in mice. Given that seminal plasma is an abundant source of transforming growth factor β, we evaluated the effect of exposure to seminal plasma on the growth of endometrial lesions. Human endometrial explants were exposed to seminal plasma or to control medium before transfer to Prkdc(scid)-mutant (severe combined immunodeficient) mice. Xenografts exposed to seminal plasma showed an eightfold increase in volume and a 4.3-fold increase in weight after 14 days. These increases were associated with increased proliferation of endometrial epithelial cells and enhanced survival and proliferation of human stromal cells compared with those in control lesions, in which human stromal cell persistence was negligible. Although the distribution of macrophages was altered, their number and activation status did not change in response to seminal plasma. Seminal plasma stimulated the production of a variety of cytokines in endometrial tissue, including growth-regulated oncogene, granulocyte macrophage colony-stimulating factor, and IL-1β. These data suggest that seminal plasma enhances the formation of endometriosis-like lesion via a direct effect on endometrial cell survival and proliferation, rather than via macrophage-mediated mechanisms. These findings raise the possibility that endometrial exposure to seminal plasma could contribute to endometriotic disease progression in women.

Complex abdominopelvic endometriosis: the radiologist's perspective.

Endometriosis is a multifocal gynecological disorder affecting approximately 6%-10% of women during their reproductive years (Giudice and Kao in: Lancet 364:1789-1799, 2004). Presenting symptomatology often relates to the anatomical structures involved. Given the complexity of both the management and pain control of patients with complex endometriosis, the British Society of Gynaecological Endoscopy has issued guidelines on the establishment of a multidisciplinary team approach to these cases (http://www.bsge.org.uk/ec-requirements-BSGE-accredited-endometriosis-centre.php). The ovaries are the most common site affected, but the gastrointestinal, genitourinary tract, chest and other soft tissues are not infrequently involved. Less well-recognized features of the disease include the deep infiltrative form of endometriosis, malignant transformation and decidualization of endometriomas under progesterone. In this pictorial essay, we will discuss the clinical presentation and review the imaging features of these complex and under appreciated forms of endometriotic disease.

Transvaginal sonographic sliding sign: accurate prediction of pouch of Douglas obliteration

Transvaginal sonographic sliding sign: accurate prediction of pouch of Douglas obliteration

Novel Molecular Strategy Tests, Issued by Laparoscopy, That Can Predict the Progression and Severity of Endometriotic Disease

Novel Molecular Strategy Tests, Issued by Laparoscopy, That Can Predict the Progression and Severity of Endometriotic Disease

MR Imaging in Deep Pelvic Endometriosis: A Pictorial Essay

Deep pelvic endometriosis is an important gynecologic disorder that is responsible for severe pelvic pain and is defined as subperitoneal invasion that exceeds 5 mm in depth. Deep pelvic endometriosis can affect the retrocervical region, uterosacral ligaments, rectum, rectovaginal septum, vagina, urinary tract, and other extraperitoneal pelvic sites. It is commonly associated with dysmenorrhea, dyspareunia, pelvic pain, urinary tract symptoms, and infertility. Because surgery remains the best therapeutic option for affected patients, the accurate preoperative assessment of the extension of endometriotic disease is extremely important. Pelvic magnetic resonance (MR) imaging is a noninvasive method with high spatial resolution that allows multiplanar evaluation of deep pelvic endometriosis and good tissue characterization, but without the use of ionizing radiation or iodinated contrast agents. MR imaging yields important findings that help grade the disease and identify subperitoneal lesion extension and other associated disease entities, thereby facilitating accurate diagnosis and adequate treatment. Radiologists should be familiar with the MR imaging findings of deep infiltrating endometriosis in various anatomic locations so that they can provide information that allows adequate presurgical counseling.

Expression and Regulation of Prostaglandin Biosynthetic Enzymes Cox-2 and Pges-1 and Receptors EP1, EP2, EP3, and EP4 in Endometriosis in the Nonhuman Primate.

Endometriosis is a debilitating disease where endometrium-like tissue forms lesions outside the uterus that result in chronic pelvic pain and infertility in 10-20% of women of reproductive age. Endometriosis is an estrogen-dependent disease and current medical therapies for the disease include oral contraceptives, aromatase inhibitors, and gonadotropin releasing hormone analogues that act by suppressing estrogen action. These therapies are not ideal in that they frequently produce undesirable menopause-like side effects such as loss of bone density and hot flushes and fail to restore fertility in women of reproductive age. Moreover, cessation of medical therapy results in reestablishment of the disease within a year in ~50-60% of patients. Our broad goal is to investigate new therapies for endometriotic disease. There is growing evidence that prostaglandin E2 (PGE2) can act to mediate the actions of steroid hormones to promote growth and development of endometriotic lesions. PGE2 biosynthesis is limited by cyclooxygenase-2 (COX-2) and prostaglandin E-synthase-1 (PGES-1) and its actions are mediated through G-protein coupled membrane receptors (EP1, EP2, EP3, and EP4). Rhesus macaques can develop endometriosis and provide a useful animal model for testing novel therapies for the disease. However, little is known regarding the PGE2 paracrine system in rhesus macaque endometrium. This study had two objectives. Our first objective was to characterize tissue specific expression on steroid hormone regulation of COX-2, PGES-1, and the EP receptors in eutopic and ectopic endometria from rhesus macaques. Our second objective was to assess the effect of COX-2, EP2 and EP4 on cell proliferation in rhesus macaque endometrial epithelial and stromal cells by pharmacological blockade. Samples were obtained from animals assigned to studies at the Oregon National Primate Research Center (ONPRC). Animal use was reviewed and approved by the ONPRC Institutional Animal Care and Use Committee. The samples included formaldehyde-fixed paraffin-embedded endometrium, spontaneous endometriosis and ectopically grafted endometrium (induced endometriosis) from castrated monkeys under estrogen treatment (n=3 each). Fresh samples of endometrium from spontaneous endometriosis were also used to prepare isolated epithelial and stromal cells cultures for in vitro study. We found that COX-2 and PGES-1 were expressed in glandular epithelia (GLE) and stroma (STR) and were significantly up regulated in ectopic compared to eutopic endometria. EP1 protein was present at similar levels in the GLE and STR of both ectopic and eutopic endometria. However, EP2 and EP4 proteins were expressed at greater levels in GLE and STR of ectopic compared to eutopic endometria. Expression of EP3 protein was very low or undetectable in both eutopic and ectopic macaque endometrium. Functional studies revealed that progesterone down-regulated EP4 but not EP2, COX-2 or PGES-1 in ectopic and eutopic endometria in vivo. Pharmacological blockade of COX-2 or EP2 and EP4 decreased proliferation of primary cultured endometrial GLE and STR. In conclusion, our data suggest that PGE2 may play an important role in the pathogenesis of endometriosis and targeting COX-2 or EP2 and EP4 could develop into novel non-estrogen or non-steroidal therapies for treatment of endometriosis in women. Supported by NIH grants HD18185 and RR000163 to ONPRC. (platform)

Peroxisome proliferator-activated receptor-(gamma) receptor ligand partially prevents the development of endometrial explants in baboons: a prospective, randomized, placebo-controlled study.

A prospective, randomized, placebo-controlled study was conducted in a baboon model to determine if a thiazolidinedione agonist of peroxisome proliferator-activated receptor-gamma, pioglitazone, can impede the development of endometriosis. Endometriosis was induced using laparoscopic, intrapelvic injection of eutopic menstrual endometrium, previously incubated with placebo or pioglitazone for 30 min, in 12 female baboons with a normal pelvis that had undergone at least one menstrual cycle since the time of captivity. At this point, the 12 baboons were randomized into two groups and treated from the day of induction. They received either PBS tablets (n = 6, placebo control, placebo tablets once a day by mouth) or pioglitazone (n = 6, test drug, 7.5 mg by mouth each day). A second and final laparoscopy was performed in the baboons to record the extent of endometriotic lesions between 24 and 42 d after induction (no difference in length of treatment between the two groups, P = 0.38). A videolaparoscopy was performed to document the number and surface area of endometriotic lesions. The surface area and volume of endometriotic lesions were significantly lower in pioglitazone treated baboons than the placebo group (surface area, 48.6 vs. 159.0 mm(2), respectively, P = 0.049; vol, 23.7 vs. 131.8 mm(3), respectively, P = 0.041). The surface area (3.5 vs. 17.8 mm(2), P = 0.017, pioglizatone vs. placebo) and overall number (1.5 vs. 9.5, P = 0.007, pioglizatone vs. placebo) of red lesions were lower in the pioglitazone group. A peroxisome proliferator-activated receptor-gamma ligand, pioglitazone, effectively reduced the initiation of endometriotic disease in the baboon endometriosis model. Using this animal model, we have shown that thiazolidinedione is a promising drug for preventive treatment of endometriosis.

Progesterone (P) resistance in the endometrium of rhesus macaques with advanced endometriosis attenuates levels of cysteine rich secretory protein 3 (CRISP3)

OBJECTIVE: Women with endometriosis often display diminished endometrial receptivity, which correlates with resistance of the endometrium to progesterone (P) action during the secretory phase. We recently reported that CRISP3 is a P-regulated protein in the macaque endometrium. Our goal was to compare CRISP3 expression in the secretory endometrium of macaques with or without endometriosis in order to better define P resistance in this animal model for endometriotic disease.