Role of Progesterone-Regulated Genes in Early Pregnancy

描述(由申请人提供)：类固醇激素孕酮(P)在妊娠的建立和维持过程中对子宫的功能有深远的影响。P的细胞作用是通过细胞内孕酮受体(PR)亚型PR-A和PR-B介导的，PR-A和PR-B是众所周知的转录因子。据推测，激素占据的PR在子宫内不同类型的细胞中触发特定基因网络的表达，这些基因的产物介导激素效应。这一建议的长期目标是确定PR调节的通路，并对其进行功能表征，这些通路是妊娠早期子宫内P反应的关键调节因子。本研究的具体目的是：1.分析DNA微阵列衍生基因在小鼠着床前子宫中PR异构体特异性调控和表达。利用寡核苷酸微阵列鉴定了几个在妊娠子宫中表达显著下调的基因，这些基因在植入时对PR拮抗剂有反应。PR异构体特异性基因敲除(KO)小鼠模型PRAKO和PRBKO将被用来识别对植入具有潜在重要性的基因。将分析这些基因在怀孕子宫中的时空表达。2.确定基因芯片衍生基因在植入前子宫中的功能作用。利用一种新开发的方法，针对选定候选基因的mRNA转录产物的反义寡核苷酸将被注射到植入前子宫中，以阻断植入期间特定基因的表达。这一干预的功能效果将得到确定。在初步研究中，反义ODN诱导的Irg1在表面上皮细胞中的表达被阻断，导致着床严重受损。用酵母双杂交方法确定Irg1在妊娠子宫中的分子靶点(S)。3.研究PR调节的蛋白水解酶抑制物p12在蜕膜组织中的作用。p12是P诱导的丝氨酸蛋白酶抑制物，在滋养层侵袭和蜕膜形成中起作用。将通过体外蛋白质相互作用方法和蛋白质组学方法鉴定妊娠子宫中p12的靶蛋白(S)。此外，还将开发一只p12KO小鼠，并对其潜在的生殖缺陷进行分析。这项拟议的研究将帮助我们确定在怀孕早期P调节胚胎-子宫相互作用的关键中介分子。

DESCRIPTION (provided by applicant): The steroid hormone progesterone (P) profoundly influences the function of the uterus during establishment and maintenance of pregnancy. The cellular actions of P are mediated through intracellular progesterone receptor (PR) isoforms, PR-A and PR-B, which are well-known transcription factors. It is postulated that hormone-occupied PR triggers the expression of specific gene networks in different cell types within the uterus and the products of these genes mediate the hormonal effects. The long-term goal of this proposal is to identify and functionally characterize the PR-regulated pathways, which are critical mediators of P response within the uterus during early pregnancy. The specific aims of this study are to: 1. Analyze PR isoform-specific regulation and expression of DNA microarray-derived genes in the preimplantation mouse uterus. Oligonucleotide microarrays were utilized to identify several genes whose expression is markedly down regulated in pregnant uterus at the time of implantation in response to a PR antagonist. The PR isoform-specific gene knock-out (KO) mouse models, PRAKO and PRBKO, will be employed to identify the genes that are potentially important for implantation. The spatio-temporal expression of these genes in the pregnant uterus will be analyzed. 2. Determine the functional roles of microarray-derived genes in the preimplantation uterus. Using a newly developed methodology, antisense oligodeoxynucleotides directed against mRNA transcripts of selected candidate genes will be administered into the preimplantation uterus to block specific gene expression during implantation. The functional effects of this intervention will be determined. In preliminary studies, antisense ODN-induced blockade of the expression of Irgl in the surface epithelium results in a severe impairment of implantation. The molecular target(s) of Irgl in the pregnant uterus will be identified by yeast two-hybrid approach. 3. Investigate the functional role of the PR-regulated protease inhibitor p12 in the decidual uterus, p12 is a serine protease inhibitor induced by P during trophoblast invasion and decidualization. The target protease(s) of p12 in the pregnant uterus will be identified by in vitro protein interaction methods and proteomics. Additionally, a p12 KO mouse will be developed and analyzed for potential reproductive defects. The proposed study will help us to identify molecules that are critical mediators of P regulation of embryo-uterine interactions during early pregnancy.