GENOME-WIDE IDENTIFICATION OF GENES REQUIRED FOR DECIDUALIZATION

 描述(申请人提供)：人类不孕不育是一个全球性的问题，胚胎植入失败是自然妊娠和体外受精过程中妊娠失败的重要原因。着床是一个极其复杂的过程，需要精确控制激素、生长因子信号和细胞-细胞接触，以协调合格胚泡和接受子宫之间的相互作用。蜕膜化是着床过程的一部分，涉及成纤维细胞样子宫内膜间质细胞的快速增殖，然后分化为上皮样蜕膜细胞，然后蜕膜细胞成为围绕着床胚胎的蜕膜组织的一部分。蜕膜化缺陷可直接导致植入失败。此外，早期蜕膜素缺陷还可能导致其他妊娠缺陷，如胎盘、胎儿宫内发育和分娩。到目前为止，由于缺乏合适的高通量筛选工具，蜕膜化过程还没有得到系统的研究。在这里，我们描述了这种体外筛选工具的产生，它包含用于蜕膜形成的荧光读数。在Aim I中，我们将使用这种体外筛选工具进行全基因组shRNA筛选，以确定蜕膜形成的新基因。在AIM II中，我们将分析蜕膜化所需转录因子的时间诱导，并建立导致蜕膜化的转录因子网络。该项目的成功完成将在植入生物学、避孕、体外受精和环境毒理学等多个研究领域产生长期影响。

 DESCRIPTION (provided by applicant): Human infertility is a global problem and failure of embryo implantation accounts for a significant percentage of pregnancy failure during both natural pregnancy and in vitro fertilization procedures. Implantation is an extremely complicated process requiring precisely controlled hormonal, growth factor signaling and cell-cell contacts which coordinate interactions between the competent blastocysts and the receptive uterus. Decidualization is part of the implantation process and involves rapid proliferation then differentiation of fibroblast-like endometrial stromal cells into epitheloid-like decidual cells whch later become part of the decidual tissue that surrounds the implanting conceptus. Decidualization defects can directly lead to implantation failure. In addition, early decidualizatin defects can also lead to other pregnancy defects such as placentation, intrauterine fetal growth, and parturition. Up to now, the process of decidualization has not been systematically studied due to the lack of a suitable high throughput screening tool. Here we describe the generation of such an in vitro screening tool that contains a fluorescent readout for decidualization. In Aim I we will use this in vitro screening tool to perform a genome-wide shRNA screen to identify novel genes for decidualization. In Aim II, we will analyze the temporal induction of transcription factors required for decidualization and establish a transcription factor network leading to decidualization. Successful completion of this project will have a long-lasting impact in multiple research fields including implantation biology, contraception, in vitro fertilization and environmental toxicology.