sirolimus and Endometriosis

Adenomyosis, endometriosis, endometritis, and typical endometrial hyperplasia are common non-cancerous diseases of the endometrium that afflict many women with life-impacting consequences. The mammalian target of the rapamycin (mTOR) pathway interacts with estrogen signaling and is known to be dysregulated in endometrial cancer. Based on this knowledge, we attempt to investigate the role of mTOR signaling in benign endometrial diseases while focusing on how the interplay between mTOR and eukaryotic translation initiation factors (eIFs) affects their development. In fact, mTOR overactivity is apparent in adenomyosis, endometriosis, and typical endometrial hyperplasia, where it promotes endometrial cell proliferation and invasiveness. Recent data show aberrant expression of various components of the mTOR pathway in both eutopic and ectopic endometrium of patients with adenomyosis or endometriosis and in hyperplastic endometrium as well. Moreover, studies on endometritis show that derangement of mTOR signaling is linked to the establishment of endometrial dysfunction caused by chronic inflammation. This review shows that inhibition of the mTOR pathway has a promising therapeutic effect in benign endometrial conditions, concluding that mTOR signaling dysregulation plays a critical part in their pathogenesis.

Topics: Adenomyosis; Endometrial Hyperplasia; Endometriosis; Endometritis; Endometrium; Female; Humans; Sirolimus; TOR Serine-Threonine Kinases; Uterine Diseases

The phosphoinositol-3 kinase (PI3K) pathway is frequently dysregulated in endometrial cancer (EC). Hormonal manipulation leads to response in some patients with EC, but resistance derived from PI3K pathway activation has been documented. Targeting mammalian target of rapamycin (mTOR) may overcome endocrine resistance. We conducted a two-institution phase II trial of everolimus and letrozole in women with recurrent EC.. Patients were considered incurable, had measurable disease, and were treated with up to two prior cytotoxic regimens. Everolimus was administered orally at 10 mg daily and letrozole was administered orally at 2.5 mg daily. Each cycle consisted of 4 weeks of therapy. Patients were treated until progression, toxicity, or complete response (CR). The primary end point was the clinical benefit rate (CBR), which was defined as CR, partial response, or stable disease (≥ 16 weeks) by RECIST 1.0 criteria. Translational studies were performed to correlate biomarkers with response.. Thirty-eight patients were enrolled (median age, 62 years; range, 24 to 82 years). Thirty-five patients were evaluable for response. The CBR was 40% (14 of 35 patients); the median number of cycles among responders was 15 (range, seven to 29 cycles). The confirmed objective response rate (RR) was 32% (11 of 35 patients; nine CRs and two partial responses; median, 15 cycles; range, eight to 29 cycles). Twenty percent of patients (seven of 35 patients) were taken off treatment after a prolonged CR and at the discretion of the treating clinician. None of the patients discontinued treatment as a result of toxicity. Serous histology was the best predictor of lack of response. Patients with endometrioid histology and CTNNB1 mutations responded well to everolimus and letrozole.. Everolimus plus letrozole results in a high CBR and RR in patients with recurrent EC. Further development of this combination in recurrent endometrioid EC is under way.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Agents; Bayes Theorem; beta Catenin; Biomarkers, Tumor; Cohort Studies; Disease-Free Survival; DNA Mutational Analysis; Endometrial Neoplasms; Endometriosis; Everolimus; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Immunosuppressive Agents; Letrozole; Middle Aged; Neoplasm Recurrence, Local; Nitriles; Phosphoinositide-3 Kinase Inhibitors; Receptors, Estrogen; Receptors, Progesterone; Sirolimus; Time Factors; TOR Serine-Threonine Kinases; Treatment Outcome; Triazoles

Neiyi Prescription of QIU (NYPQ) is a traditional Chinese medicine prescription for the treatment of endometriosis (EMS). Here, we aimed to examine the effects and mechanisms of NYPQ on angiogenic ability in EMS.. EMS rats were established with estradiol valerate and autologous transplantation. EMS rats were intraperitoneally injected with chloroquine (CQ, 40 mg/kg), rapamycin (RAPA, 1 mg/kg), and monoclonal antibody VEGF (anti-VEGF, 3 mg/g/d) or administered 5, 10, 20 mg/g/d NYPQ decoction through oral gavage for 4 weeks, respectively. By the before and end of the treatment period, the volume of the endometriotic lesions was measured. The pathological morphology, angiogenesis, and the number of autophagosomes of the endometriotic lesion were observed by hematoxylin and eosin staining, immunohistochemistry, and transmission electron microscope, respectively. The cell viability, apoptosis, and angiogenesis of HUVECs were detected by MTT, flow cytometry, and lumen formation experiment, respectively. The expression levels of VEGF, autophagy-/apoptosis-/PPARγ/NF-κB- pathway-related proteins in endometrium tissues or HUVECs were detected by western blot assays.. The autophagy agonist rapamycin reduced the lesion size, the microvessel density, and VEGF expression, and promoted the production of autophagosomes and the expression of autophagy-related proteins, while the autophagy inhibitor chloroquine had the opposite effects. In vivo, NYPQ could dose-dependently reduce lesion volume and microvessel density, ameliorate histopathological features and promote autophagosome production of ectopic endometrium. Moreover, serum-containing NYPQ could significantly inhibit the cell viability and tube formation of HUVECs and elevate HUVECs apoptosis. Besides, NYPQ significantly reduced VEGF and promoted autophagy-/apoptosis-related protein expressions. Also, NYPQ might promote autophagy and inhibit angiogenesis by activating the PPARγ/NF-κB pathway.. Collectively, these findings indicate that NYPQ has therapeutic potential in experimentally induced peritoneal endometriosis, and its mechanism may be related to the activation of the PPARγ/NF-κB signaling pathway.

Topics: Animals; Autophagy; Chloroquine; Drugs, Chinese Herbal; Endometriosis; Female; Humans; Neovascularization, Pathologic; NF-kappa B; PPAR gamma; Prescriptions; Rats; Signal Transduction; Sirolimus

To explore the mechanism for the role of autophagy in endometriosis, and to provide a theoretical basis for prevention and treatment of endometriosis.
 Methods: The endometrial CRL-7566 cells were treated with ATG5 siRNA, autophagic activator rapamycin and autophagic inhibitor 3-MA, respectively. The cell proliferation and invasion were detected by clonal formation, cell growth curve and MTT assay. The clinical specimens of endometriosis were collected from 20 cases. The expression of autophagy marker LC3II and autophagy substrate protein P62 were detected.
 Results: Rapamycin inhibited the proliferation and clonal formation of CRL-7566 cells, while autophagy inhibitor 3-MA and ATG5 siRNA showed opposite effect. Moreover, rapamycin inhibited filopodia growth in endometriosis, whereas overexpression of filopodia-relevant protein fascin-1 inhibited the decrease in invasiveness caused by rapamycin. In clinical samples, we also found a significant decrease of LC3II while an increase in P62 compared with the control group.
 Conclusion: Autophagy inhibition may contribute to an increase in endometrial cell proliferation and invasiveness. Autophagy activation could be a potential strategy for endometriosis therapy.. 目的：探讨自噬在子宫内膜异位症中的作用机制，为通过靶向干预自噬防治子宫内膜异位症提供理论基础。方法：使用ATG5干扰片段、自噬激活剂雷帕霉素(rapamycin)和自噬抑制剂3-MA等干预子宫内膜CRL-7566细胞自噬，通过克隆形成、细胞生长曲线和MTT检测其对细胞的生长增殖和侵袭力的影响。收集20例子宫内膜异位症患者临床标本，检测自噬标志物LC3II和自噬底物蛋白P62的表达。结果：雷帕霉素抑制子宫内膜CRL-7566细胞的增殖和克隆形成，而自噬抑制剂3-MA及自噬相关基因ATG5的干扰作用相反。雷帕霉素抑制子宫内膜细胞的伪足生长，而伪足相关蛋白fascin-1的过表达能抑制雷帕霉素引起的侵袭力下降。与对照组相比，在子宫内膜异位症患者的临床标本中，自噬标志物LC3II显著降低而自噬底物P62增加。结论：自噬抑制可能促进子宫内膜细胞的增殖和侵袭，自噬激活可能是子宫内膜异位症治疗的潜在靶点。.

Topics: Autophagy; Carrier Proteins; Cell Line; Cell Proliferation; Endometriosis; Endometrium; Female; Gene Expression Regulation; Humans; Microfilament Proteins; Microtubule-Associated Proteins; RNA-Binding Proteins; Sirolimus

Mammalian target of rapamycin (mTOR) is known to be a major negative regulator of autophagy. Recent studies have shown that mTOR activity is abnormally increased in endometriotic lesions. In endometriosis, abnormal mTOR activity may contribute to the alteration of endometrial cell autophagy, which may affect apoptosis because endometrial cell autophagy is directly involved in the regulation of apoptosis. To test this hypothesis, we investigated whether endometrial cell autophagy is altered by aberrant mTOR activity and is associated with apoptosis in ovarian endometriotic cysts. Our results show that endometrial cell autophagy induction was increased by mTOR inhibition as the menstrual cycle progresses in the normal endometrium, and that it is correlated with apoptosis. However, in endometriotic tissues from ovarian endometriotic cysts, autophagy, mTOR activity and apoptosis were constant throughout the menstrual cycle, suggesting that a constant level of autophagy is maintained by disinhibition of mTOR activity during the menstrual cycle in endometriotic tissues and is related to decreased apoptosis. Indeed, compared with normal endometrium, increased mTOR activity during the secretory phase in endometriotic tissues inhibited autophagy and apoptosis induction. In addition, to determine the direct effect of autophagy induction mediated by mTOR on endometriotic cell apoptosis, endometriotic cells were treated with rapamacin (mTOR inhibitor) with and without 3-methyladenine (3-MA, autophagy inhibitor). Although rapamycin treatment induced autophagy and led to apoptosis promotion, the pro-apoptotic effect of rapamycin was reversed by the addition of 3-MA, suggesting that mTOR inhibition promotes endometriotic cell apoptosis via autophagy induction. In conclusion, our results suggest that aberrant mTOR activity in ovarian endometriotic cysts leads to alteration of endometrial cell autophagy, which is associated with abnormal apoptosis.

Topics: Adenine; Animals; Apoptosis; Autophagy; Cysts; Endometriosis; Endometrium; Female; Gene Expression Regulation; Humans; Menstrual Cycle; Primary Cell Culture; Sirolimus; Stromal Cells; TOR Serine-Threonine Kinases

Deep infiltrating endometriosis (DIE) is a particular clinical and histological entity of endometriosis responsible for chronic pelvic pain and infertility. Here we characterize the proliferative phenotype of DIE cells, to explore the cellular and molecular mechanisms that could explain their aggressive potential. In addition, the inhibition of mTOR/AKT pathway was tested, as a potential treatment of DIE. Included were 22 patients with DIE and 12 control patients without endometriosis. Epithelial and stromal cells were extracted from biopsies of eutopic endometrium and deep infiltrating endometriotic nodules from patients with DIE. Cell proliferation was determined by thymidine incorporation. Oxidative stress was assayed by spectrofluorometry. The ERK and mTOR/AKT pathways were analyzed in vitro by Western blot and for AKT in vivo in a mouse model of DIE. The proliferation rate of eutopic endometrial cells and of deep infiltrating endometriotic cells from DIE patients was higher than that of endometrial cells from controls. The hyperproliferative phenotype of endometriotic cells was associated with an increase in endogenous oxidative stress, and with activation of the ERK and mTOR/AKT pathways. mTOR/AKT inhibition by temsirolimus decreased endometriotic cell proliferation both in vitro and in vivo in a mouse model of DIE. Blocking the mTOR/AKT pathway offers new prospects for the treatment of DIE.

Topics: Adult; Animals; Biopsy; Cell Proliferation; Disease Models, Animal; Endometriosis; Endometrium; Female; Humans; Mice; Mice, Nude; Middle Aged; Oxidative Stress; Phenotype; Protein Kinase Inhibitors; Reactive Oxygen Species; Sirolimus; Spectrometry, Fluorescence; TOR Serine-Threonine Kinases

Rapamycin is a widely used drug with antifungal, immunosuppressant and antiangiogenic effects. Herein, we studied whether immunosuppressive doses of rapamycin are capable of influencing endometriotic lesions.. We tested in vitro the potential of rapamycin to inhibit endothelial cell sprouting using the aortic ring assay and we further studied its effect on the expression of proliferating cell nuclear antigen (PCNA), apoptotic cell death-associated activated caspase-3 and vascular endothelial growth factor (VEGF) in cultured endometrial tissue fragments. In addition, we analyzed the drug in vivo after induction of endometriotic lesions by transplanting isolated endometrial fragments into the dorsal skinfold chamber of Syrian golden hamsters. Using intravital fluorescence microscopy, we repetitively analyzed angiogenesis, neovascularization and microcirculatory parameters over a time period of 14 days in rapamycin-treated animals and DMSO-treated controls.. Administration of rapamycin significantly reduced the size of the endometriotic lesions. This was associated by inhibition of VEGF-mediated angiogenesis as indicated by a suppression of endothelial cell sprouting in vitro and a reduction of microvessel density in endometriotic lesions in vivo. Moreover, rapamycin directly inhibited cell proliferation within endometrial tissue, while manifestation of apoptotic cell death remained unaffected.. Our data indicate that administration of rapamycin may represent a novel therapeutic approach for an antiangiogenic treatment of endometriosis.

Topics: Animals; Aorta; Cell Proliferation; Cricetinae; Endometriosis; Female; Immunosuppressive Agents; Mesocricetus; Microscopy, Fluorescence; Neovascularization, Pathologic; Sirolimus

To expand on a preliminary study comparing the chemotactic potential of peritoneal fluid (PF) from women with and without endometriosis and to characterize this activity via immunosuppressants and a protease.. Case control study.. University center.. Fifty-nine women with endometriosis and 44 without, undergoing laparoscopy.. Collection of PF, endometriotic, ovarian, and endometrial biopsies at laparoscopy.. Chemotactic activity of PF was tested via an in vitro assay alone and in the presence of immunosuppressants cyclosporin A (CSA), FK506, rapamycin, and type XVII-b(S-V8) protease and in media incubated with endometriotic, ovarian, or endometrial biopsy specimens.. The PF from women with endometriosis had significantly greater chemotactic activity (cells per well, mean +/- SD) than without endometriosis (142 +/- 39 versus 48 +/- 17). Cyclosporin A significantly inhibited the chemotactic activity of the endometriotic PF; FK506 and rapamycin did not. Incubation of media with endometriotic tissue, but not ovarian or endometrial, for > or = 7 hours displayed chemotactic activity. Protease type XVII-b(S-V8) added to endometriotic PF inhibited this chemotactic activity.. Peritoneal fluid from patients with endometriosis contains a protein chemotactic factor attracting inflammatory cells into the peritoneal cavity, possibly secreted by endometriotic implants. This chemotactic factor may be a member of the immunophilin family because of its inhibition profile.

Topics: Ascitic Fluid; Case-Control Studies; Chemotaxis; Cyclosporine; Endometriosis; Female; Humans; Immunosuppressive Agents; Macrophages; N-Formylmethionine Leucyl-Phenylalanine; Polyenes; Proteins; Serine Endopeptidases; Sirolimus; Tacrolimus