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（由应用提供）：人类不育症是一个全球问题，胚胎植入的失败占自然怀孕和体外受精程序期间妊娠失败的很大一部分。植入是一个极其复杂的过程，需要精确控制的激素，生长因子信号传导和细胞 - 细胞接触，该过程协调了主管胚泡和相关子宫之间的相互作用。固定化是植入过程的一部分，涉及快速的增殖，然后将成纤维细胞样子宫基质细胞分化为上皮样细胞，后来成为植入概念的决定组织的一部分。义词化缺陷可以直接导致植入失败。此外，早期的决定性缺陷也可能导致其他妊娠缺陷，例如胎盘，胎儿内胎儿生长和分娩。到目前为止，由于缺乏合适的高吞吐量筛选工具，因此尚未系统地研究决定性化的过程。在此提案中，我们建议使用最近开发的最先进的CRISPR/CAS基因组编辑技术生成如此急需的工具。在目的中，我将使用CRISPR来生成双重培养基细胞系，通过将两个荧光记者击倒到内源性标记的局部局部局部，以确定化。在AIM II中，我们将使用该新工具对可以干扰决策的化合物进行小规模的高吞吐量筛选。该项目的成功完成将对多个研究领域产生持久影响，包括植入生物学， 避孕，体外受精和环境毒理学。