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）：类固醇激素黄体酮（P）在怀孕建立和维持过程中深刻影响子宫的功能。 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，激素占据的 PR 会触发子宫内不同细胞类型中特定基因网络的表达，并且这些基因的产物介导激素效应。该提案的长期目标是确定 PR 调节途径并对其进行功能表征，这些途径是妊娠早期子宫内 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 DNA 微阵列衍生基因的 PR 异构体特异性调控和表达。利用寡核苷酸微阵列来鉴定一些基因，这些基因的表达在怀孕子宫中在植入时响应于 PR 拮抗剂而显着下调。 PR亚型特异性基因敲除（KO）小鼠模型PRAKO和PRBKO将用于鉴定对植入可能重要的基因。将分析这些基因在怀孕子宫中的时空表达。 2. 确定微阵列衍生基因在植入前子宫中的功能作用。使用新开发的方法，针对选定候选基因的 mRNA 转录本的反义寡脱氧核苷酸将被注射到植入前的子宫中，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 Irgl 表达阻断导致植入严重受损。妊娠子宫中 Irgl 的分子靶标将通过酵母双杂交方法进行鉴定。 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是滋养层侵袭和蜕膜化过程中由P诱导的丝氨酸蛋白酶抑制剂。将通过体外蛋白质相互作用方法和蛋白质组学来鉴定妊娠子宫中 p12 的靶蛋白酶。此外，还将开发 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识别在妊娠早期胚胎-子宫相互作用的 P 调节中起关键调节作用的分子。