AN IN VITRO SCREENING TOOL FOR DECIDUALIZATION

一种用于分化的体外筛选工具

• 批准号: 8969886
• 负责人: Liang Ma
• 金额: $ 19.06万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-03 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8969886
• 关键词: Accounting; Biology; Birth; Cell Line; Cells; Chemicals; Clinic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Conceptus; Contraceptive Agents; Contraceptive methods; Decidual Cell; Decidual Cell Reactions; Decidual Reaction; Defect; Development; Endocrine disruption; Endometrial Stromal Cell; FDA approved; Failure; Fertility Agents; Fertilization in Vitro; Fetal Growth; Fibroblasts; Foundations; Genes; Genomics; Growth Factor; Hormonal; Human; IGFBP1 gene; Implant; In Vitro; Infertility; Lead; Libraries; Modification; Pharmaceutical Preparations; Placentation; Play; Polyploidy; Preclinical Drug Evaluation; Pregnancy; Procedures; Process; Reporter; Reproduction; Research; Role; Technology; Tissues; Uterus; blastocyst; drug development; environmental chemical; environmental toxicology; failure Implantation; genome editing; high throughput screening; human embryonic stem cell; implantation; intercellular communication; natural Blastocyst Implantation; novel; public health relevance; reproductive; research study; screening; small molecule libraries; tool

 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. In this proposal, we propose to generate such much-needed tool using the recently developed state-of-the-art CRISPR/Cas genome editing technology. In Aim I we will use CRISPR to generate a dual-reporter cell line for decidualization by knocking two fluorescent reporters into endogenous loci of decidual markers. In Aim II, we will use this new tool to perform a small scale high throughput screening for chemical compounds that can interfere with 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. .

 描述（由申请人提供）：人类不育是一个全球性问题，胚胎植入失败占自然妊娠和体外受精过程中妊娠失败的很大比例。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，以协调有能力的囊胚和接受子宫之间的相互作用。蜕膜化是着床过程的一部分，包括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，后者后来成为围绕着床孕体的蜕膜组织的一部分。蜕膜化缺陷可直接导致着床失败。此外，早期蜕膜化缺陷也可导致其他妊娠缺陷，如胎盘形成、胎儿宫内生长和分娩。由于缺乏合适的高通量筛选工具，迄今为止尚未对蜕膜化过程进行系统的研究。在这项提案中，我们建议使用最近开发的最先进的CRISPR/Cas基因组编辑技术来生成这种急需的工具。在目的I中，我们将使用CRISPR通过将两个荧光报告基因敲入蜕膜标志物的内源基因座来产生用于蜕膜化的双报告基因细胞系。在Aim II中，我们将使用这种新工具来进行小规模的高通量筛选，以筛选可以干扰蜕膜化的化合物。该项目的成功完成将对多个研究领域产生长期影响，包括植入生物学， 避孕、体外受精和环境毒理学。.