CHEMOKINE REGULATION IN MODELS OF ENDOMETRIOSIS

子宫内膜异位症是一种常见的妇科疾病， 痛经、盆腔疼痛和生育能力下降。患病率估计值 范围从2- 50%，尽管大多数子宫内膜异位症学者认为， 它发生在大约10%的育龄美国妇女中。的 美国每年因子宫内膜异位症造成的医疗费用超过 十亿美元最近的研究表明，子宫内膜异位症植入物激活 局部腹膜炎症反应介导临床 症状我们假设招募和随后的 腹腔内活化巨噬细胞的积聚是一种 子宫内膜异位症的早期和必要的步骤 并确定了炎症和血管生成的浓度升高 结果：1999年12月至2000年12月， 子宫内膜异位症我们建议研究合成的调节， 一种代表性趋化因子RANTES的分泌， 单核细胞和T细胞的化学引诱物。我们证明了RANTES 位于正常子宫内膜的间质室中， 子宫内膜异位症植入物和mRNA和蛋白质都表达， 子宫内膜间质细胞而非上皮细胞。具体目标 #1我们将使用高度纯化（>95%）的原代基质细胞培养物， 比较RANTES在正常人的在位和异位细胞中的产生 受试者，患有子宫内膜异位症的女性和患有不明原因的女性 不孕我们的初步数据表明，RANTES蛋白分泌 前两种情况不同。在具体目标#2中，我们将研究 天然卵巢类固醇激素（雌二醇， 孕酮），拮抗剂（他莫昔芬，RU 486，达那唑）和其他 细胞因子（TNF-α）、IL-1 α和β、干扰素-γ）来调节 RANTES在mRNA和蛋白水平的体外表达。的表达 含有477个碱基对的人RANTES基因启动子的载体 将使用克隆的荧光素酶报告基因上游来绘制 瞬时转染子宫内膜的转录调控基序 和子宫内膜异位症基质细胞。具体目标#3将在 与Osteen博士合作，使用他的人体体内模型， 移植于裸鼠腹腔内。激素 以及在体外上调和下调免疫反应性RANTES的细胞因子 将被施用给携带人子宫内膜植入物的小鼠， 确定这些化合物是否调节人RANTES在完整组织中， vivo.同源抗激素或细胞因子中和抗体将被免疫。 在一些实施方案中，施用RANTES以确认RANTES调节作用是特异性的。 RANTES和其他趋化因子很可能在早期发挥作用， 在伴随这一过程的炎症过程中， 综合征这些分子应该为未来提供理想的靶点 用于医学治疗的新型治疗性拮抗剂的开发 子宫内膜异位症

Endometriosis is a common human gynecologic disorder associated with dysmenorrhea, pelvic pain and reduced fertility. Prevalence estimates range from 2-50%, although most scholars of endometriosis believe that it occurs in approximately 10% of reproductive aged American women. The annual United States health costs attributable to endometriosis exceed $1 billion. Recent studies suggest that endometriosis implants activate local peritoneal inflammatory responses that mediate the clinical symptoms. We hypothesize that the recruitment and subsequent accumulation of activated macrophages in the peritoneal cavity was an early and requisite step in the establishment of endometriosis implants and identified elevated concentrations of inflammatory and angiogenic cytokines (RANTES, IL-6, IL-8, VEGF) in peritoneal fluid of women with endometriosis. We propose to investigate the regulation of synthesis and secretion of one representative chemokine, RANTES, a potent chemoattractant for monocytes and T cells. We demonstrated that RANTES is localized in the stromal compartment of normal endometrium and endometriosis implants and that both mRNA and protein are expressed in endometrial stromal but not epithelial cells in vitro. In Specific Aim #1 we will use highly purified (>95%) primary stromal cell cultures to compare RANTES production in eutopic and ectopic cells from normal subjects, women with endometriosis and women with unexplained infertility. Our preliminary data indicate that RANTES protein secretion differs in the former two conditions. In Specific Aim #2, we will study the ability the natural ovarian steroid hormones (estradiol, progesterone), antagonists (tamoxifen, RU486, danazol) and other cytokines (TNF-alpha), IL-1alpha and beta, interferon-gamma) to modulate RANTES expression in vitro at the mRNA and protein levels. An expression vector containing 477 base pairs of the human RANTES gene promoter cloned upstream of a luciferase reporter will be used to map the transcriptional regulatory motifs in transiently transfected endometrial and endometriosis stromal cells. Specific Aim #3 will be executed in collaboration with Dr. Osteen using his in vivo model of human endometrium transplanted into the nude mouse peritoneal cavity. Hormones and cytokines that up- and down-regulate immunoreactive RANTES in vitro will be administered to mice bearing human endometrial implants to determine if these compounds regulate human RANTES in intact tissues in vivo. Cognate anti-hormones or cytokine neutralizing antibodies will be administered to confirm that the RANTES modulating effects are specific. It is likely that RANTES and other chemokines play early, requisite play early, requisite roles in the inflammatory process that accompanies this syndrome. These molecules should provide ideal targets for the future development of novel therapeutic antagonists for the medical treatment of endometriosis.