Molecular Basis of Treating Endometriosis by Prostaglandin E2 Receptor Inhibitors

描述（由申请人提供）：子宫内膜异位症是一种雌激素依赖性疾病。目前的药物治疗抑制雌激素的生物合成和行动不能防止疾病的再次发生，并危及育龄妇女的妊娠成功。这表明迫切需要确定子宫内膜异位症非雌激素治疗靶点的潜在细胞信号通路。然而，前列腺素E2 （PGE2）促进子宫内膜异位症的生存；强调的分子机制在很大程度上是未知的。我们的长期目标是了解子宫内膜异位症发病机制中PGE2生物合成和信号串扰的分子和细胞方面，以确定新的靶向治疗方法。本应用程序的目的是了解PGE2信号通路在子宫内膜异位症的生存和生长。我们的中心假设是PGE2受体EP2和EP4的功能丧失抑制子宫内膜异位症的生存和生长。特异性Aim-1将决定EP2和EP4功能缺失诱导子宫内膜异位症细胞凋亡的机制。特异性Aim- 2将确定EP2和ep4介导的PGE2信号在子宫内膜异位症中免疫调节和增强巨噬细胞吞噬能力的机制。特异性Aim-3将确定EP2和EP4功能丧失减少子宫内膜异位症雌激素产生的机制。我们的实验方法包括：(i)基因组和药理学抑制EP2和EP4；（ii）稳定的荧光标记的人子宫内膜异位症上皮细胞、基质细胞、巨噬细胞以及来自子宫内膜异位症患者的异位和异位子宫内膜，（iii）裸和Rag2g(c)小鼠异种移植模型，(v)基于分子、细胞、生化和显微镜的检测；（六）全动物生物成像方法。其基本原理是，成功完成拟议的研究将为我们的知识基础提供一个缺失的和基本的元素，否则我们就无法理解通过选择性抑制EP2和EP4诱导人类子宫内膜异位症细胞凋亡的机制。此外，预期结果将推进目前对子宫内膜异位症发病机制的认识，并增加对PGE2信号在子宫内膜异位症生存中的理解。获得这些知识是至关重要的，可以转化为治疗患有子宫内膜异位症的妇女。我们预计选择性抑制EP2和EP4可通过多种机制诱导子宫内膜异位症细胞凋亡，增强浸润性巨噬细胞自身的吞噬能力，减少子宫内膜异位症细胞雌激素的产生。我们的研究结果将具有临床影响，因为它将允许首次开发新的和急需的治疗策略，以抑制EP2和EP4信号作为新的非雌激素靶点，用于治疗育龄妇女子宫内膜异位症。这是一项R21申请，旨在解决NIH/NICHD在妇女生殖健康方面的任务。

DESCRIPTION (provided by applicant): Endometriosis is an estrogen dependent disease. Current medical therapies to inhibit estrogen biosynthesis and actions fail to prevent reoccurrence of the disease and compromise success of pregnancy in child-bearing age women. This suggests a crucial need to identify potential cell signaling pathways for nonestrogen therapeutic targets for endometriosis. Prostaglandin E2 (PGE2) promotes survival of endometriosis, however; the underlined molecular mechanisms are largely unknown. Our long-term goal is to understand molecular and cellular aspects of PGE2 biosynthesis and signaling cross-talk in the pathogenesis of endometriosis in order to identify new targeted therapies. The objective of this application is to understand PGE2 signaling pathways in survival and growth of endometriosis. Our central hypothesis is that loss-of-function of PGE2 receptors EP2 and EP4 inhibits survival and growth of endometriosis. Specific Aim-1 will determine the mechanisms through which loss-of-function of EP2 and EP4 induces apoptosis of endometriosis. Specific Aim- 2 will determine the mechanisms through which EP2 and EP4-mediated PGE2 signaling immunomodulate and enhance the phagocytic ability of macrophages in endometriosis. Specific Aim-3 will determine the mechanisms through which loss-of-function of EP2 and EP4 decrease estrogen production in endometriosis. Our experimental approaches include: (i) genomic and pharmacological inhibition of EP2 and EP4; (ii) stable fluorescence-labeled human endometriotic epithelial cells, stromal cells, macrophages, and eutopic and ectopic endometria from endometriosis patients, (iii) nude and Rag2g(c) mice xenograft models, (v) molecular, cellular, biochemical, and microscopy-based assays; and (vi) whole animal bioimaging method. The rationale is that successful completion of the proposed research will contribute a missing and fundamental element to our base of knowledge without which the mechanism through which selective inhibition of EP2 and EP4 induces apoptosis of human endometriotic cells cannot be understood. In addition, the expected results will advance the current knowledge of the pathogenesis of endometriosis and increase the understanding of PGE2 signaling in survival of endometriosis. The acquisition of such knowledge is critical and could be translated to treat women suffering from endometriosis. It is our expectation that selective inhibition of EP2 and EP4 will induce apoptosis of endometriotic cells, increase phagocytic ability of infiltrated macrophages in endometriosis per se, and decrease estrogen production by the endometriotic cells through multiple mechanisms. Our findings would have clinical impact because it would allow for the first time to develop new and much needed therapeutic strategies to inhibit EP2 and EP4 signaling as novel nonestrogen targets for the treatment of endometriosis in child-bearing age women. This is a R21 application addresses the mission of NIH/NICHD on women's reproductive health. PUBLIC HEALTH RELEVANCE: The objective of the proposed research is to determine molecular and cellular mechanisms through which selective inhibition of prostaglandin E2 (PGE2) receptors EP2 and EP4 inhibits survival of endometriosis in order to identify PGE2 as potential nonestrogen or nonsteroidal target for the treatment of endometriosis in women. The expected outcomes of this project are that selective inhibition of EP2 and EP4-mediated PGE2 signaling will: (i) inhibit survival of endometriosis through suppressed cell survival and activated intrinsic apoptotic pathways; (ii) increase phagocytic ability of infiltrated peritoneal macrophages around endometriosis per se; (iii) inhibit local estrogen production in endometriosis; and (iv) provide exciting new knowledge that fill the gap in understanding of the pathogenesis of endometriosis. The novel findings of this project are expected to establish potential translational opportunities for treatment of endometriosis by blocking EP2 and EP4 receptors and could emerge as potential nonestrogen or nonsteroidal therapy for endometriosis in child- bearing age women.