Generation of a Model to Study Uterine Gland Function

项目总结 不孕不育和流产是影响女性的常见健康疾病。子宫腺有 在生育、发育不良和疾病中发挥重要的生物学作用。妊娠丢失是最常见的 在子宫中观察到人类妊娠的并发症，以及反复妊娠丢失和不孕。 腺体基因敲除动物模型。子宫腺上皮(GE)及其分泌物 在子宫容受性、胚泡/胚胎存活和着床方面具有生物学作用 间质细胞蜕膜化--建立和维持 怀孕了。然而，由于缺乏合适的小鼠，子宫腺生物学研究受到阻碍。 有条件地删除或过度表达成年子宫GE中的基因的模型。这个 这项提案的目标就是解决这个问题。Prs29(蛋白酶、丝氨酸29或植入丝氨酸 蛋白水解酶基因2)是一个在青春期后在成人的GE中强势而特异地表达的基因 小鼠子宫和其他雌性生殖道组织中不存在。在目标1中，改进了Cre重组酶 使用CRISPR/Cas9基因组将(Icre)插入到Prss 29基因的3‘非翻译区 工程学。ICre的活性和细胞特异性表达将在发育和 成年组织用rosa26报告小鼠。目标2是建立和验证Prss 29的有用性- ICRE小鼠用于研究成年子宫腺功能。具体表示叉头盒a2(Foxa2) 在成年小鼠和人子宫的GE中具有潜在的生物学作用 妊娠期卵巢类固醇激素反应基因与GE功能Prss 29-iCre小鼠将 用于有条件地删除成人子宫中的Foxa2，并确定其对生育和 怀孕了。Prss 29-iCre小鼠将与RiboTag小鼠结合用于主动分离 翻译的mRNAs，这将被测序，以揭示怀孕的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 探索/发展研究资助计划(家长R21)“，鼓励研究资助 支持项目早期和概念阶段的应用程序。在这些结束时 研究，我们希望能产生一种新的小鼠模型，可用于子宫腺的研究 生物学、再生和疾病，填补了我们对子宫腺现有知识的一个重大空白 在怀孕期间的功能。

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.