Effects of a Histone Methyltransferase Inhibitor on Fertility on a Rat Model of Endometriosis

Endometriosis is an estrogen-dependent, inflammatory disease that affects 5-10% of women of reproductive age. It is defined as the growth of functioning endometrium outside the uterus that results in severe pelvic pain and often infertility. Currently, endometriosis has no cure, and available treatments have limited efficacy and side effects. Epigenetics play a key role in the etiology of this disease, and we have previously shown that treatment with histone methyltransferase inhibitors (HMTi) in an animal model of endometriosis significantly decreases vesicle development, suggesting the potential use of epigenetic drugs for endometriosis. The objective of this study was to investigate the effects of HMTi on fertility by analyzing the effects of the drug on expression of fertility genes in this model. Endometriosis induction was performed in female Sprague Dawley rats. Two weeks after, rats were treated intraperitoneally with HMTi for four weeks. At sacrifice, uterine tissues were collected and mRNA extracted to study fertility gene expression using a real-time polymerase chain reaction (RT2 Profiler PCR Array). HMTi treatment modified the expression of a limited number of genes (2 out of 84), and increased the expression of key genes related to embryonic implantation and development of the ovary. These observations suggest that HTMi has a positive effect on fertility, a possibility that requires additional investigations in vivo

Elucidating the Expression Profile of EZH2 Isoforms in Endometriosis: An immunohistochemical study

Endometriosis is an estrogen-dependent gynecological disease that affects 1 out of 10 women of reproductive age causing severe pelvic pain and infertility. Factors including genetics, environment, inflammation, and recently epigenetics have been shown to play roles in the pathophysiology of this disease. Histone methylation is an epigenetic modification that modulates gene expression by causing changes in the chromatin structure. Trimethylation of histone 3 at lysine residue 27 (H3K27me3) is a histone mark related to gene repression. EZH2 is the histone methyltransferase (HMT) responsible of catalyzing H3K27me3. It has been shown that the EZH2 is involved in carcinogenesis; however, the specific role of EZH2 in endometriosis is unknown. This is important because there are drugs available that block this enzyme’s functioning, and could serve as a potential new treatment. We have previously shown that endometriotic lesions are characterized by high H3K27me3 nuclear immunostaining. Therefore, we hypothesize that EZH2 will be highly expressed in lesions compared to endometrium of patients and controls. Using immunohistochemistry (IHC) of an endometriosis Tissue Micro Array (TMA), EZH2α and β nuclear intensity were assessed using specific antibodies. We observed that pelvic endometriotic lesions (peritoneal and fallopian tube) have higher EZH2α intensity scores compared to control tissues. EZH2β nuclear immunostaining analysis is ongoing. We expect to observe a higher nuclear intensity score in endometriotic lesions compared to endometriosis free endometria. This study is the first to analyze the expression profile of EZH2 isoforms in endometriosis. These studies will help better understand the role of EZH2 in this disease

A polymorphism in a let-7 microRNA binding site of KRAS in women with endometriosis.

Endometriosis is found in 5-15% of women of reproductive age and is more frequent in relatives of women with the disease. Activation of KRAS results in de novo endometriosis in mice, however, activating KRAS mutations have not been identified in women. We screened 150 women with endometriosis for a polymorphism in a let-7 microRNA (miRNA) binding site in the 3'-UTR of KRAS and detected a KRAS variant allele in 31% of women with endometriosis as opposed to 5% of a large diverse control population. KRAS mRNA and protein expression were increased in cultured endometrial stromal cells of women with the KRAS variant. Increased KRAS protein was due to altered miRNA binding as demonstrated in reporter assays. Endometrial stromal cells from women with the KRAS variant showed increased proliferation and invasion. In a murine model, endometrial xenografts containing the KRAS variant demonstrated increased proliferation and decreased progesterone receptor levels. These findings suggest that an inherited polymorphism of a let-7 miRNA binding site in KRAS leads to abnormal endometrial growth and endometriosis. The LCS6 polymorphism is the first described genetic marker of endometriosis risk

Efficacy of an environmental enrichment intervention for endometriosis: a pilot study

IntroductionWe have previously shown that Environmental Enrichment (EE), a multi-modal psychosocial intervention consisting of increased social interaction, novelty, and open spaces, improved disease presentation, anxiety, and immune-related disturbances in the rat model of endometriosis. However, there is a knowledge gap regarding the effects of EE interventions in patients with this painful, inflammatory chronic disease.AimTo adapt and test the efficacy of an EE intervention on pelvic pain, mental health, perceived stress, quality of life, and systemic inflammation in endometriosis patients through a randomized clinical trial (RCT).Materials and methodsA multidisciplinary team with expertise in physiology, neuroscience, psychology, and women’s health adapted and implemented a two-arm RCT comparing an EE intervention with a wait-list control group. Six EE modules administered on alternate weeks were provided to patients in the intervention (N = 29); controls received education only. Survey data and biospecimens were collected at baseline, end-of-study, and 3-months post-intervention to assess pain (Brief Pain Inventory, BPI), endometriosis-related quality of life-QoL (Endometriosis Health Profile-30, EHP30), anxiety (Generalized Anxiety Disorder 7, GAD7), depression (Patient Health Questionnaire for Depression 8, PHQ8), pain catastrophizing (Pain Catastrophizing Score, PCS), stress (Perceived Stress Scale-14, PSS14), and saliva cortisol levels (AM, PM).ResultsCompared to the wait-list controls, participants in the EE intervention showed significantly decreased GAD-7 scores at the end of the intervention and 3-month follow-up. Depression, perceived stress, and QoL improved at the 3-month follow-up compared to baseline. While pain levels did not improve, they significantly correlated with anxiety, depression, QoL and pain catastrophizing scores.ConclusionThis pilot RCT demonstrated significant improvements in anxiety and depressive symptoms, QoL, and perceived stress, supporting enriched environments as an integrative psychosocial intervention to be used as adjuvant to the standard of care for endometriosis pain

Expression and Function of Nuclear Receptor Coactivator 4 Isoforms in Transformed Endometriotic and Malignant Ovarian Cells

Iron is proposed to contribute to the transition from endometriosis to specific subtypes of ovarian cancers (OVCAs). Regulation of intracellular iron occurs via a ferritinophagic process involving NCOA4 (Nuclear Receptor Coactivator 4), represented by two major isoforms (NCOA4α and NCOA4β), whose contribution to ovarian cancer biology remains uninvestigated. We thus generated transformed endometriotic cells (via HRASV12A, c-MYCT58A, and p53 inactivation) whose migratory potential was increased in response to conditioned media from senescent endometriotic cells. We identified elevated NCOA4 mRNA in transformed endometriotic cells (relative to non-transformed). Knockdown of NCOA4 increased ferritin heavy chain (FTH1) and p21 protein which was accompanied by reduced cell survival while NCOA4β overexpression reduced colony formation. NCOA4α and NCOA4β mRNA were elevated in malignant versus non-malignant gynecological cells; NCOA4α protein was increased in the assessed malignant cell lines as well as in a series of OVCA subtypes (relative to normal adjacent tissues). Further, NCOA4 protein expression was regulated in a proteasome- and autophagy-independent manner. Collectively, our results implicate NCOA4 in ovarian cancer biology in which it could be involved in the transition from precursors to OVCA

Compositions and methods for treating endometriosis

Provided herein are methods of treating and/or preventing endometriosis or symptom thereof, assays for diagnosing/prognosing endometriosis, compositions and formulations for treating and/or preventing endometriosis or symptom thereof, and populations of endometiotic cells, including life-extended populations of cells

Noninvasive diagnosis of endometriosis: Review of current peripheral blood and endometrial biomarkers

A noninvasive biomarker-based test could help shorten the diagnostic delay for endometriosis. The most investigated biomarker sources are peripheral blood and endometrium. Discovery of endometriosis biomarkers is often hypothesis-driven, i.e. when one or a few biomarkers are investigated based on their role in the disease pathogenesis. Alternatively, a hypothesis-generating approach has been followed using the "omics" technologies. A variety of biomarkers for endometriosis have been investigated, but no biomarker has been validated for clinical use. Many challenges lie ahead in the endometriosis biomarker field. In the future, harmonized collection and reporting methods should allow large-scale international collaboration for highly powered studies.status: publishe

Impact of coping strategies on quality of life of adolescents and young women with endometriosis

Purpose: Endometriosis is a hormone-dependent, inflammatory, painful condition affecting 1 in 10 women during their reproductive years. The symptoms of endometriosis—dysmenorrhea, dyspareunia, infertility—negatively impact the quality of life (QoL) of the affected women. Few studies have been conducted on mental health and QoL impact in a younger endometriosis patient population (adolescents and young women). This study quantitative, cross-sectional study was designed to address this gap by ascertaining whether coping strategies may impact the QoL of this patient population. Methods: After consent, participants (n = 24) completed a sociodemographic questionnaire, Beck Anxiety Inventory (BAI), Beck Depression Inventory II (BDI-II), Coping Strategies Inventory (CSI), Endometriosis Health Patient-5 (EHP-5) and Visual Analogue Scale (VAS). Results: Participants reported the use of both positive and maladaptive strategies to deal with the symptomatology, which were associated with QoL levels and mental health status. Associations between QoL and maladaptive coping strategies (e.g. autocriticism, social withdrawal) were uncovered. Cognitive restructuring was identified as an adaptive coping strategy that impacts QoL positively. Conclusion: These results provide additional evidence showing that endometriosis symptoms substantially affect the psychological well-being of young patients and identify opportunities for interventions (e.g. cognitive behavioral, rational/emotive therapy) to implement coping styles leading to improved QoL

Environmental Manipulations as an Effective Alternative Treatment to Reduce Endometriosis Progression

Treatments for endometriosis include pharmacological or surgical procedures that produce significant side effects. We aimed to determine how environmental enrichment (EE) could impact the progression of endometriosis using the autotransplantation rat model. Female rats were exposed to EE (endo-EE: toys and nesting materials, 4 rats per cage, larger area enclosure) or no enrichment (endo-NE: 2 rats per cage) starting on postnatal day 21. After 8 weeks, sham surgery or surgical endometriosis was induced by suturing uterine horn tissue next to the intestinal mesentery, then allowed to progress for 60 days during which EE or NE continued. At the time of killing, we measured anxiety behaviors, collected endometriotic vesicles and uterus, and processed for quantitative real-time polymerase chain reaction for corticotropin-releasing hormone (CRH), urocortin-1, CRH receptors type 1 and type 2, and glucocorticoid receptor (GR). Endometriosis did not affect anxiety-like behaviors, yet rats in enriched conditions showed lower basal anxiety behaviors than the nonenriched group. Importantly, the endo-EE group showed a 28% reduction in the number of endometriosis vesicles and the vesicles were significantly smaller compared to the endo-NE group. Endometriosis increased CRH and GR only in the vesicles of endo-NE, and this increase was dampened in the endo-EE. However, urocortin 1 was increased in the vesicles of the endo-EE group, suggesting different pathways of activation of CRH receptors in this group. Our results suggest that the use of multimodal complementary therapies that reduce stress in endometriosis could be an effective and safe treatment alternative, with minimal side effects