Role of miR15a and miR34c in PGE2 Signaling in the Pathogenesis of Endometriosis

 描述(申请人提供)：子宫内膜异位症是一种育龄妇女的雌激素依赖型和黄体酮抵抗型炎症性疾病，其特征是子宫腔外出现功能性子宫内膜。这种神秘疾病的主要症状是月经痛、慢性盆腔疼痛和不孕不育。因此，它是美国育龄妇女子宫切除的唯一主要原因，每年估计社会成本约为694亿美元。由于不良副作用和未能防止复发，目前的抗雌激素疗法只能开出很短的时间。为了开发治疗子宫内膜异位症的新的分子靶向治疗方法，需要确定抑制子宫内膜异位症病变生长和存活的新的信号通路。子宫内膜异位症患者腹腔液中前列腺素E_2(PGE_2)浓度高于无疾病女性，这种升高被认为促进了子宫内膜异位症的生存、生长和炎症。在人类子宫内膜异位症病变中，miR15a和miR34c的表达似乎是表观遗传沉默。我们的主要假设是选择性地抑制PGE2受体EP2和EP4通过改变DNA甲基化和组蛋白修饰来逆转miR15a和miR34c的表观遗传沉默，并且联合抑制EP2和EP4以及激活miR15a和miR34c协同和协同地抑制子宫内膜异位症的生长和生存。特异性AIM-1将确定选择性抑制PGE2受体EP2和EP4与激活miR15a和miR34c对子宫内膜异位症生长和生存的联合治疗效果。特异性AIM-2将确定miR15a和miR34c介导PGE2受体EP2和EP4抑制子宫内膜异位症生长和存活的分子和细胞机制。特异性AIM-3将建立抑制PGE2受体EP2和EP4逆转子宫内膜异位症表观遗传沉默和恢复miR15a和miR34c转录激活的分子机制和表观遗传途径。实验方法包括人源化的RAG2γ(C)小鼠模型，活化人子宫内膜异位上皮和间质细胞，通过药理学和基因组学方法抑制EP2和EP4，激活miR15a和miR34c，全动物生物成像，以及表观遗传学、分子生物学和显微镜技术。该研究项目的成功完成有望为通过抑制EP2和EP4，激活miR15a和miR34c来治疗子宫内膜异位症提供新的基础知识，为制定基于表型的个性化非类固醇联合治疗提供新的基础知识。这种新的非激素疗法可以为现有的激素疗法提供一种更有效的替代疗法，并有望成为子宫内膜异位症治疗的未来创新转变。同样重要的是，该项目中确定的潜在治疗方法可以推断为其他炎症性疾病。这项申请涉及NICHD关于妇女生殖健康的任务。

 DESCRIPTION (provided by applicant): Endometriosis is an estrogen-dependent and progesterone-resistant inflammatory disease of reproductive-age women characterized by the presence of functional endometrium outside the uterine cavity. The major symptoms of this enigmatic disease are painful menstruation, chronic pelvic pain, and infertility. Thus, it represents the single major cause for hysterectomy in reproductive-age women in United States, with annual estimated societal cost of ~$69.4 billion. Current anti-estrogen therapies can be prescribed only for a short time because of undesirable side effects and failure to prevent recurrence. Identification of novel signaling pathways that inhibit growth and survival of endometriotic lesions is needed to develop new molecular-targeted therapies for endometriosis. Concentrations of prostaglandin E2 (PGE2) in the peritoneal fluid are higher in endometriosis women compared to disease-free women, and this increased PGE2 is thought to promote the survival, growth, and inflammation of endometriosis. Expression of miR15a and miR34c appears to be epigenetically silenced in endometriotic lesions in human patients. Our overarching hypothesis is that selective inhibition of PGE2 receptors EP2 and EP4 reverses epigenetic silencing of miR15a and miR34c by altering DNA methylation and histone modifications in endometriosis and that combinatorial inhibition of EP2 and EP4 and activation of miR15a and miR34c cooperatively and synergistically inhibit growth and survival of endometriosis. Specific Aim-1 will determine the combinatorial therapeutic effects of selective inhibition of PGE2 receptors EP2 and EP4 and activation of miR15a and miR34c on growth and survival of endometriosis. Specific Aim-2 will identify the molecular and cellular mechanisms through which miR15a and miR34c mediate inhibitory effects of PGE2 receptors EP2 and EP4 on growth and survival of endometriosis. Specific Aim-3 will establish the molecular mechanisms and epigenetic pathways through which inhibition of PGE2 receptors EP2 and EP4 reverses epigenetic silencing and restores transcriptional activation of miR15a and miR34c in endometriosis. The experimental approaches such as the humanized Rag2γ(c) mice models, active human endometriotic epithelial and stromal cells, inhibition of EP2 and EP4 and activation of miR15a and miR34c by pharmacologic and genomic approaches, whole animal bioimaging, and epigenetics, molecular biology and microscopy-based techniques will be used. Successful completion of the research project is expected to provide new fundamental knowledge to formulate personalized phenotype-based combinatorial nonsteroidal therapy through inhibition of EP2 and EP4 and activation of miR15a and miR34c for endometriosis. This novel non-hormonal therapy could provide a more effective alternative treatment to existing hormonal therapies and hold promise for future innovative shifts in endometriosis treatment. Equally important, potential treatments identified in this project can be extrapolated to other inflammatory diseases. This application addresses missions of NICHD on women's reproductive health.