Progesterone Action in the Endometrium of Women with Endometriosis

描述（由申请人提供）：子宫内膜异位症导致美国2 - 8%的妇女疼痛和/或不孕，每年花费超过200亿美元。尽管子宫内膜异位症的临床和经济影响，其发病机制和病理生理仍然知之甚少，治疗选择仍然有限。我们和其他人已经证明了子宫内膜异位症妇女在位子宫内膜中受孕酮（P）调节的基因的异常表达，表明对P的抵抗。鉴于P抑制子宫内膜增殖，为月经时的凋亡细胞死亡奠定基础，并诱导胚胎着床的接受性，P抵抗可能通过促进子宫内膜种植体的增殖和存活，通过抑制胚胎植入。目前的知识差距包括正常子宫内膜功能所需的P作用的量和持续时间，子宫内膜异位症妇女对P的需求可能改变的程度，以及P抵抗的功能后果。我们已经开始使用一种新的人体月经周期的体内模型来定义未受影响的女性的P作用和需求，在该模型中，通过实验来定义循环P浓度。在生育受试者中的初步数据验证了我们的方法，定义了正常结构（~2-3 ng/mL）和功能（~5-10 ng/mL）子宫内膜分化所需的近似最低阈值血清P浓度，并确定了可作为P作用剂量敏感性标志物的mRNA种类。根据初步资料和现有文献，我们假设：（1）子宫内膜异位症患者在P浓度达到正常分化时，子宫内膜结构和/或功能发育异常;（2）超生理P浓度可克服孕激素抵抗;（3）P抵抗性间质中异常的旁分泌信号导致了子宫内膜上皮基因表达的异常模式。为了检验前两个假设，我们将使用我们建立的体内模型，直接比较P的要求正常子宫内膜分化的生育妇女与不孕妇女和子宫内膜异位症。将在组织学上评估P对子宫内膜结构的影响，并将通过qRT-PCR和分离的上皮和基质细胞组分的微阵列分析以及组织切片的免疫染色来评估P的功能影响。为了验证第三个假设，我们将联合收割机正常上皮细胞与来自患有或不患有子宫内膜异位症的妇女的基质细胞结合作为小鼠宿主的异种移植物。用雌激素和不同剂量的P治疗将允许评估间质对异常子宫内膜miRNA、mRNA和蛋白质表达的贡献。这些研究将直接评估子宫内膜的P抵抗力，间质对异常子宫内膜功能的影响，提供P作用的剂量特异性标志物，并建立有和无子宫内膜异位症的生育和不孕妇女的P需求。这些人类数据将为开发新的子宫内膜异位症诊断和治疗策略提供坚实的基础。

DESCRIPTION (provided by applicant): Endometriosis causes pain and/or infertility in 2 - 8% of women in the U.S., with an annual cost of more than $20 billion. Despite the clinical and financial impact of endometriosis, its pathogenesis and pathophysiology remain poorly understood and treatment options remain limited. We and others have demonstrated abnormal expression of genes regulated by progesterone (P) in eutopic endometrium of women with endometriosis, suggesting resistance to P. Given that P inhibits endometrial proliferation, sets the stage for apoptotic cell death at menstruation, and induces receptivity to embryo implantation, P resistance could contribute to the pathogenesis and pathophysiology of endometriosis by facilitating proliferation and survival of endometrial implants and by inhibiting embryo implantation. Current knowledge gaps include the amount and duration of P action required for normal endometrial function, the extent to which P requirements may be altered in women with endometriosis, and the functional consequences of P resistance. We have begun to define P action and requirements in unaffected women using a novel in vivo model of the human menstrual cycle, in which circulating P concentrations are defined experimentally. Preliminary data in fertile subjects validates our approach, defines the approximate minimum threshold serum P concentrations required for normal structural (~2-3 ng/mL) and functional (~5-10 ng/mL) endometrial differentiation, and identifies mRNA species that can serve as dose-sensitive markers of P action. Base on preliminary data and existing literature, we hypothesize: (1) that women with endometriosis exhibit abnormal endometrial structural and/or functional development at P concentrations that achieve normal differentiation in fertile controls, (2) that progesterone resistance can be overcome by supra-physiological P concentrations; and (3) that abnormal patterns of endometrial epithelial gene expression in affected women result from abnormal paracrine signaling in P-resistant stroma. To test the first two hypotheses, we will use our established in vivo model to directly compare P requirements for normal endometrial differentiation in fertile women to those in infertile women with and without endometriosis. The effects of P on endometrial structure will be assessed histologically and the functional effects of P will be assessed by qRT-PCR and microarray analysis of isolated epithelial and stromal cell fractions and immunostaining of tissue sections. To test the third hypothesis, we will combine normal epithelial cells with stromal cells from women with or without endometriosis as a xenograft in a murine host. Treatments with estrogen and varying doses of P will permit assessment of the stromal contribution to abnormal endometrial miRNA, mRNA, and protein expression. These studies will assess directly endometrial P resistance, the effect of stroma on abnormal endometrial function, provide dose-specific markers of P action, and establish P requirements in fertile and infertile women with and without endometriosis. These human data will provide a solid foundation for the development of novel diagnostic and therapeutic strategies for endometriosis.