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: 7098758
• 负责人: Indrani C Bagchi
• 金额: $ 30.25万
• 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/7098758
• 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）：类固醇激素黄体酮(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激素受体（PR）异构体PR- a和PR- b介导的，它们是众所周知的转录因子。据推测，激素占据的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的作用。本研究的长期目标是确定pr调控通路的功能特征，这些通路是妊娠早期子宫内P反应的关键介质。本研究的具体目的是：1。分析植入前小鼠子宫中PR亚型特异性调控和DNA微阵列衍生基因的表达。利用寡核苷酸微阵列鉴定了几个基因，这些基因的表达在植入时对PR拮抗剂的反应中在妊娠子宫中明显下调。PR亚型特异性基因敲除（KO）小鼠模型PRAKO和PRBKO将用于鉴定对着床可能重要的基因。我们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植入前子宫中的功能作用。使用一种新开发的方法，针对选定候选基因的mRNA转录物的反义寡脱氧核苷酸将被施用于着床前子宫，以阻断着床期间特定基因的表达。这种干预的功能效果将被确定。在初步研究中，反义odn诱导的表面上皮中Irgl表达的阻断导致了植入的严重损害。利用酵母双杂交方法确定Irgl在妊娠子宫中的分子靶点。3. 研究pr调控的蛋白酶抑制剂p12在蜕膜子宫中的功能作用，p12是P在滋养细胞侵袭和蜕膜过程中诱导的丝氨酸蛋白酶抑制剂。通过体外蛋白相互作用方法和蛋白质组学方法鉴定妊娠子宫中p12的靶蛋白酶。此外，将开发p12 KO小鼠并分析其潜在的生殖缺陷。该研究将帮助我们确定在妊娠早期胚胎-子宫相互作用中P调节的关键介质分子。