Progesterone Action in the Endometrium of Women with Endometriosis

描述(申请人提供)：在美国，子宫内膜异位症会导致2-8%的女性疼痛和/或不孕，每年的成本超过200亿美元。尽管子宫内膜异位症对临床和经济有影响，但其发病机制和病理生理机制仍然知之甚少，治疗选择仍然有限。我们和其他人在子宫内膜异位症患者的在位内膜中发现了孕酮(P)调节基因的异常表达，提示对孕酮的抵抗。鉴于孕酮抑制子宫内膜的增殖，为月经时的细胞凋亡奠定基础，并诱导胚胎着床的容受性，孕酮抵抗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和病理生理过程。目前的知识空白包括正常子宫内膜功能所需的磷作用量和持续时间，子宫内膜异位症患者的磷需求可能发生变化的程度，以及磷抵抗的功能后果。我们已经开始使用一种新的人体月经周期活体模型来定义未受影响的女性的磷作用和需求，在该模型中，循环中的磷浓度是通过实验确定的。在有生育能力的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)和功能性(~5-10 ng/mL)子宫内膜分化所需的大约最低血清P浓度，并确定了可作为P作用的剂量敏感标志物的mRNA种类。根据初步数据和现有文献，我们假设：(1)子宫内膜异位症患者在P浓度下表现出异常的子宫内膜结构和/或功能发育，在生育对照组中实现正常分化；(2)超生理P浓度可以克服孕激素抵抗；(3)受影响妇女子宫内膜上皮基因异常表达模式是由于耐P间质中异常的旁分泌信号所致。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妇女和患有和不患有子宫内膜异位症的不育妇女对正常子宫内膜分化的磷需求。P对子宫内膜结构的影响将通过组织学评估，P的功能效应将通过qRT-PCR和分离的上皮和间质细胞组分的微阵列分析和组织切片的免疫染色来评估。为了验证第三个假设，我们将正常的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相结合，作为小鼠宿主的异种移植。用雌激素和不同剂量的P进行治疗，可以评估间质对子宫内膜miRNA、mRNA和蛋白异常表达的贡献。这些研究将直接评估子宫内膜的磷抵抗，间质对子宫内膜功能异常的影响，提供磷作用的剂量特异性标记物，并确定患有和不患有子宫内膜异位症的生育和不育妇女的磷需求。这些人类数据将为开发治疗子宫内膜异位症的新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.