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

 描述（由申请人提供）：子宫内膜异位症是育龄妇女的一种雌激素依赖性和孕酮耐药的炎性疾病，其特征是子宫腔外存在功能性子宫内膜。这种神秘疾病的主要症状是月经痛、慢性盆腔痛和不孕。因此，它是美国育龄妇女子宫切除术的唯一主要原因，每年估计社会成本约为694亿美元。目前的抗雌激素疗法只能在短时间内使用，因为它有不良的副作用，而且不能防止复发。需要鉴定抑制子宫内膜异位症病变生长和存活的新信号通路，以开发新的子宫内膜异位症分子靶向疗法。子宫内膜异位症妇女的腹膜液中前列腺素E2（PGE 2）的浓度高于无疾病妇女，这种增加的PGE 2被认为是促进子宫内膜异位症的生存，生长和炎症。miR 15 a和miR 34 c的表达似乎在人类患者的肿瘤病变中表观遗传学沉默。我们的首要假设是，选择性抑制PGE 2受体EP 2和EP 4通过改变子宫内膜异位症中的DNA甲基化和组蛋白修饰来逆转miR 15 a和miR 34 c的表观遗传沉默，并且组合抑制EP 2和EP 4以及激活miR 15 a和miR 34 c协同和协同抑制子宫内膜异位症的生长和存活。特异性目标-1将确定选择性抑制PGE 2受体EP 2和EP 4以及激活miR 15 a和miR 34 c对子宫内膜异位症的生长和存活的组合治疗效果。特异性目的-2将确定miR 15 a和miR 34 c介导PGE 2受体EP 2和EP 4对子宫内膜异位症生长和存活的抑制作用的分子和细胞机制。特异性Aim-3将建立分子机制和表观遗传途径，通过抑制PGE 2受体EP 2和EP 4逆转表观遗传沉默并恢复子宫内膜异位症中miR 15 a和miR 34 c的转录激活。将使用的实验方法包括人源化Rag 2 γ（c）小鼠模型、活性人增生上皮和基质细胞、通过药理学和基因组方法抑制EP 2和EP 4以及激活miR 15 a和miR 34 c、全动物生物成像以及基于表观遗传学、分子生物学和显微镜的技术。该研究项目的成功完成有望为通过抑制EP 2和EP 4以及激活miR 15 a和miR 34 c来制定个性化的基于表型的组合非甾体治疗子宫内膜异位症提供新的基础知识。这种新型的非激素疗法可以为现有的激素疗法提供更有效的替代治疗，并为子宫内膜异位症治疗的未来创新转变带来希望。同样重要的是，该项目中确定的潜在治疗方法可以外推到其他炎症性疾病。这项申请涉及国家排雷中心在妇女生殖健康方面的任务。

 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.