Role of Progesterone-Regulated Genes in Early Pregnancy

黄体酮调控基因在妊娠早期的作用

• 批准号: 6772480
• 负责人: Indrani C Bagchi
• 金额: $ 30.98万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-07 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6772480
• 关键词: embryo implantation; female; genetic regulation; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; laboratory mouse; messenger RNA; microarray technology; northern blottings; pregnancy; progesterone; progesterone receptors; protease inhibitor; protein protein interaction; proteomics; receptor expression; trophoblast; uterus; yeast two hybrid system

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.

描述（由申请人提供）：类固醇激素孕酮（P）在妊娠建立和维持期间深刻影响子宫功能。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，它们是众所周知的转录因子。据推测，子宫内占据的PR触发了子宫内不同细胞类型中特定基因网络的表达，这些基因的产物介导了激素效应。这项建议的长期目标是确定和功能特性的PR调节途径，这是在子宫内的P反应在妊娠早期的关键介质。本研究的具体目的是：1.分析PR亚型特异性调控和DNA微阵列衍生基因在着床前小鼠子宫中的表达。寡核苷酸微阵列被用来确定几个基因的表达显着下调，在怀孕的子宫在植入时，响应PR拮抗剂。将采用PR亚型特异性基因敲除（KO）小鼠模型PRAKO和PRBKO来鉴定对植入可能重要的基因。将分析这些基因在妊娠子宫中的时空表达。2.确定微阵列衍生基因在着床前子宫中的功能作用。使用一种新开发的方法，针对选定的候选基因的mRNA转录的反义寡脱氧核苷酸将被施用到植入前子宫中，以阻断植入期间的特定基因表达。将确定这种干预的功能效果。在初步研究中，反义ODN诱导的表面上皮中Irgl表达的阻断导致植入的严重损害。将通过酵母双杂交方法鉴定妊娠子宫中Irgl的分子靶标。3.研究PR调节的蛋白酶抑制剂p12在蜕膜子宫中的功能作用，p12是P诱导的丝氨酸蛋白酶抑制剂，在滋养层细胞侵袭和蜕膜化过程中发挥作用。将通过体外蛋白质相互作用方法和蛋白质组学鉴定妊娠子宫中p12的靶蛋白酶。此外，将开发p12 KO小鼠并分析其潜在的生殖缺陷。拟议的研究将帮助我们确定的分子是关键的P调节胚胎-子宫的相互作用在妊娠早期。