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）：类固醇激素孕酮（P）深刻影响子宫在妊娠的建立和维持期间的功能。 P的细胞作用是通过细胞内孕酮受体（PR）同工型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，激素占用的P​​R会触发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作用。该提案的长期目标是识别和在功能上表征PR调节的途径，PR调节的途径是妊娠早期子宫内P反应的关键介体。这项研究的具体目的是：1。分析植入前小鼠子宫中DNA微阵列衍生基因的PR同工型特异性调节和表达。利用寡核苷酸微阵列来鉴定几种基因，它们的表达在植入时响应PR拮抗剂时在孕妇的子宫中明显下降。将采用PR同工型特异性基因敲除（KO）小鼠模型Prako和Prbko，以识别对植入可能重要的基因。这些基因在怀孕子宫中的时空表达将进行分析。 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mRNA转录本的反义寡脱氧核苷酸将用于植入前子宫中，以阻止植入过程中特定的基因表达。将确定此干预的功能效应。在初步研究中，反义ODN诱导的表面上皮中IRGL表达的封锁会导致严重的植入损害。孕妇子宫中IRGL的分子靶标将通过酵母双杂交方法鉴定。 3。研究PR调节的蛋白酶抑制剂p12在牙本子宫中的功能作用，p12是P12在滋养细胞侵袭和decIDAILIDIAD期间由P诱导的丝氨酸蛋白酶抑制剂。孕妇子宫中p12的靶蛋白酶将通过体外蛋白质相互作用方法和蛋白质组学鉴定。另外，将开发并分析P12 KO小鼠的潜在生殖缺陷。拟议的研究将帮助我们确定妊娠早期胚胎相互作用的P调节的关键介体的分子。