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%的女性疼痛和/或不孕，每年的费用超过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实，在子宫内膜异位症女性的异位子宫内膜中，孕酮(P)调控的基因表达异常，提示对P有耐药性。鉴于P抑制子宫内膜增殖，为月经时凋亡细胞死亡奠定基础，并诱导胚胎着床的接受性。P抗性可能通过促进子宫内膜植入物的增殖和存活以及抑制胚胎着床而参与子宫内膜异位症的发病机制和病理生理。目前的知识空白包括正常子宫内膜功能所需的P作用量和持续时间，子宫内膜异位症女性对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体内模型来确定未受影响妇女的P作用和需求，其中循环P浓度是通过实验确定的。可育受试者的初步数据验证了我们的方法，确定了正常结构（~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作用剂量敏感标记的mRNA物种。根据初步数据和现有文献，我们假设：(1)子宫内膜异位症女性在P浓度达到正常分化的情况下，子宫内膜结构和/或功能发育异常；(2)孕酮抵抗可以通过超生理P浓度克服；(3)患者子宫内膜上皮基因表达模式异常是由耐p间质旁分泌信号异常引起的。为了验证前两个假设，我们将使用我们建立的体内模型，直接比较有生育能力的女性与有或没有子宫内膜异位症的不孕女性正常子宫内膜分化的P要求。P对子宫内膜结构的影响将通过组织学评估，P的功能影响将通过qRT-PCR和分离上皮细胞和基质细胞部分的微阵列分析以及组织切片的免疫染色来评估。为了验证第三个假设，我们将正常上皮细胞与来自患有或不患有子宫内膜异位症的女性的基质细胞结合，作为异种移植物移植到小鼠宿主体内。用雌激素和不同剂量的P治疗可以评估基质对子宫内膜异常miRNA、mRNA和蛋白质表达的影响。这些研究将直接评估子宫内膜P抵抗，基质对子宫内膜异常功能的影响，提供P作用的剂量特异性标志物，并确定有或无子宫内膜异位症的可育和不育妇女的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.