Generation of a Model to Study Uterine Gland Function

研究子宫腺功能的模型的生成

• 批准号: 9360767
• 负责人: THOMAS E SPENCER
• 金额: $ 19.19万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-28 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9360767
• 关键词: 3' Untranslated Regions; Address; Adult; Affect; Animal Model; Applications Grants; Bioinformatics; Biological; Biology; CRISPR/Cas technology; Cell model; Cells; Characteristics; Complication; Conceptus; Decidual Cell Reactions; Development; Developmental Biology; Diagnosis; Disease; Dysplasia; Endometrial adenocarcinoma; Endometrium; Enterobacteria phage P1 Cre recombinase; Epithelium; Female; Fertility; Generations; Genes; Genetic Models; Genome engineering; Genomics; Gland; Goals; Health; Hormone Responsive; Human; Infertility; Knock-in Mouse; Knowledge; Messenger RNA; Modeling; Mus; NIH Program Announcements; Natural regeneration; Ovarian Steroid Hormone; Parents; Pathway interactions; Peptide Hydrolases; Pregnancy; Pregnancy Complications; Pregnancy Maintenance; Pregnancy loss; Prevention; Process; Puberty; Publishing; Recurrence; Regulation; Regulatory Element; Reporter; Reproduction; Research; Research Personnel; Research Project Grants; Research Proposals; Role; Serine; Serine Protease; Stromal Cells; Study models; Tissues; Training; Translating; United States National Institutes of Health; Uterine Cancer; Uterine Gland; Uterus; Woman; Women' s Health; blastocyst; endometriosis; functional genomics; human tissue; implantation; improved; infertility treatment; interest; knockout animal; mouse model; negative affect; novel; overexpression; prevent; programs; recombinase; reproductive tract; success; transcription factor; transcriptome; transcriptome sequencing; uterine receptivity; uterus endometriosis

PROJECT SUMMARY Infertility and pregnancy loss are common health disorders affecting women. Uterine glands have important biological roles in fertility, dysplasia and disease. Pregnancy loss is the most common complication of human gestation, and recurrent pregnancy loss and infertility are observed in uterine gland knockout animal models. Uterine glandular epithelia (GE) and, by inference, their secretions have biological roles in uterine receptivity, blastocyst/conceptus survival and implantation, and stromal cell decidualization- all essential processes for the establishment and maintenance of pregnancy. However, uterine gland biology research is hampered by the lack of suitable mouse models to conditionally delete or overexpress genes specifically in the GE of the adult uterus. The goal of this proposal is to address that problem. Prss29 (protease, serine 29 or implantation serine proteinase gene two) is a gene expressed robustly and specifically after puberty in the GE of the adult mouse uterus and not in other female reproductive tract tissues. In Aim 1, improved Cre recombinase (iCre) will be inserted into the 3’ untranslated region of the Prss29 gene using CRISPR/Cas9 genome engineering. Activity and cell-specific expression of iCre will be determined in the developing and adult tissues using Rosa26 reporter mice. Aim 2 is to establish and validate the usefulness of Prss29- iCre mice for study of adult uterine gland function. Forkhead box a2 (Foxa2) is specifically expressed in the GE of the adult mouse and human uterus and has a potential biological role in regulation of ovarian steroid hormone responsive genes and GE function during pregnancy. Prss29-iCre mice will be used to conditionally delete Foxa2 in the adult uterus and determine its impact on fertility and pregnancy. Prss29-iCre mice will be used in conjuction with RiboTag mice to isolate actively translated mRNAs, which will be sequenced to uncover the GE active transcriptome of pregnancy. This application specifically targets NIH program announcement PA-13-303 entitled “NIH Exploratory/Developmental Research Grant Program (Parent R21)” that encourages research grant applications to support the early and conceptual stages of a project. At the conclusion of these studies, we expect to have generated a novel mouse model useful for the study of uterine gland biology, regeneration, and disease and fill a significant gap in our existing knowledge of uterine gland function during pregnancy.

项目摘要 不育和怀孕丧失是影响女性的常见健康障碍。子宫腺具有 在生育，发育不良和疾病中重要的生物学作用。怀孕是最常见​​的 在子宫中观察到人妊娠的并发症以及复发性怀孕丧失和不育 腺体敲除动物模型。子宫腺上皮（GE），并通过推断它们的分泌物 在子宫受体，胚泡/概念的生存和植入中具有生物学作用，以及 基质细胞决策 - 建立和维护的所有基本过程 怀孕。但是，子宫腺生物学研究受到缺乏合适的小鼠的阻碍 在成年子宫的GE中有条件删除或过表达基因的模型。 该建议的目标是解决该问题。 Press29（蛋白酶，串行29或植入串行 蛋白酶基因二）是一个基因，在成年人的GE中富有稳健和特别表达 小鼠子宫，而不是其他女性生殖道组织。在AIM 1中，改进的CRE重组酶 （ICRE）将使用CRISPR/CAS9基因组插入Press29基因的3'未翻译区域 工程。 ICRE的活性和细胞特异性表达将在发育和 使用ROSA26报告基因小鼠的成年组织。目标2是建立和验证Press29-的实用性 ICRE小鼠研究成年子宫腺功能。叉子盒A2（FOXA2）专门表示 在成年小鼠和人子宫的GE中，在调节中具有潜在的生物学作用 怀孕期间卵巢类固醇马酮反应性基因和GE功能。 PRSS29-icre小鼠会 用于在成年子宫中有条件删除FOXA2，并确定其对生育能力的影响 怀孕。 Press29-icre小鼠将与Ribotag小鼠一起使用，以主动分离 翻译的mRNA将测序以发现妊娠的GE主动转录组。 该应用程序专门针对NIH计划公告PA-13-303标题为“ NIH 鼓励研究赠款的探索/发展研究赠款计划（父母R21） 支持项目的早期和概念阶段的应用。在这些结束时 研究，我们希望产生一种新型的小鼠模型，可用于研究子宫腺 生物学，再生和疾病，并填补了我们对子宫腺的现有知识的显着空白 怀孕期间的功能。

项目成果

Generation of Mouse for Conditional Expression of Forkhead Box A2.

• DOI: 10.1210/en.2018-00158
• 发表时间: 2018-04
• 期刊: Endocrinology
• 影响因子: 4.8
• 作者: Peng Wang;San-pin Wu;K. Brooks;A. M. Kelleher;Jessica Milano-Foster;F. DeMayo;T. Spencer