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.

项目概要 不孕症和流产是影响女性的常见健康疾病。 在生育、发育不良和疾病中发挥重要的生物学作用。 子宫内观察到人类妊娠并发症、反复流产和不孕 子宫腺上皮（GE）及其分泌物（通过敲除推断） 在子宫容受性、囊胚/概念存活和着床中具有生物学作用，并且 基质细胞蜕膜化——建立和维持基质细胞的所有重要过程 然而，由于缺乏合适的小鼠，子宫腺生物学研究受到阻碍。 模型有条件地删除或过度表达特定于成人子宫 GE 中的基因。 该提案的目标是解决 Prss29（蛋白酶、丝氨酸 29 或植入丝氨酸）。 蛋白酶基因2)是青春期后在成人GE中强烈且特异表达的基因 目标 1 中，改进了 Cre 重组酶。 (iCre) 将使用 CRISPR/Cas9 基因组插入 Prss29 基因的 3' 非翻译区 iCre 的活性和细胞特异性表达将在开发和研究中确定。 使用 Rosa26 报告小鼠的成人组织目标 2 是建立并验证 Prss29- 的有效性。 用于研究成人子宫腺功能的 iCre 小鼠特异表达 Forkhead box a2 (Foxa2)。 在成年小鼠和人类子宫的 GE 中，并且在调节中具有潜在的生物学作用 Prss29-iCre 小鼠在怀孕期间的卵巢类固醇激素反应基因和 GE 功能。 用于有条件地删除成人子宫中的 Foxa2 并确定其对生育力和生育力的影响 Prss29-iCre 小鼠将与 RiboTag 小鼠联合使用以主动隔离。 翻译的 mRNA，将被测序以揭示妊娠的 GE 活性转录组。 该应用程序专门针对 NIH 计划公告 PA-13-303，标题为“NIH 探索性/发展性研究资助计划（母版 R21）”鼓励研究资助 在项目结束时支持项目的早期和概念阶段的应用程序。 研究中，我们期望生成一种可用于子宫腺研究的新型小鼠模型 生物学、再生和疾病，填补了我们现有子宫腺知识的重大空白 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