A new animal model for investigating the invasive trophoblast lineage

项目摘要/摘要 胎盘在妊娠期间起着至关重要的作用，保证胎儿的生长发育。成功 怀孕取决于胎盘对母体环境的适应。人类胎盘被归类为 血液病，特点是子宫内滋养细胞广泛侵袭。在这个过程中， 妊娠、侵袭性滋养细胞或绒毛外滋养细胞，如人类所称，从 胎盘进入子宫实质，在那里它们将胎盘固定在子宫上，重塑子宫 螺旋动脉。子宫血管重塑是为胎儿和正常胎儿提供足够的营养流的核心 胎儿发育。滋养层细胞群体的组织和结构异常与 患有妊娠疾病的人。浅层滋养细胞侵袭与子宫血管异常改变 与先兆子痫、早孕丢失、宫内发育受限、早产和 胎盘早剥。另一方面，滋养层细胞过度侵袭，如植入性胎盘， 危及怀孕、产妇健康和未来生育。触发分化的分子事件 侵袭性滋养层细胞谱系和滋养层引导的子宫螺旋动脉重塑较差 明白了。已有实验工具可用于确定侵袭性滋养层细胞的候选调控因子 并测试它们的生理相关性。大鼠胎盘形成的特点是广泛的宫内 滋养层侵袭和滋养层引导的螺旋动脉重塑类似发育过程 在人类胎盘中很明显。大鼠的全球基因组编辑是研究亚群的有效工具 涉及调控深层胎盘的候选基因，但并非所有调控深层胎盘的候选基因 胎盘摆放。有些基因在胚胎发育中具有多种作用，因此无法评估特定的作用。 在侵袭性滋养层细胞谱系中。条件等位基因的产生绕过了这个问题。在 建议的研究，我们将产生一种大鼠模型，用于产生侵袭性滋养细胞系特异性条件 使用Crispr/Cas9基因组编辑的等位基因。在本项目中，我们建立了一个表达Cre的转基因大鼠模型 Pr17b1调控序列在侵袭性滋养层细胞中的重组酶及验证 Cre司机大鼠品系。拟议中的实验将为科学界提供有价值的新工具 候选基因在调控滋养层侵袭和滋养层引导的子宫螺旋中的作用 动脉重塑，因此是探索高危妊娠病因学的新的实验范式。

PROJECT SUMMARY/ABSTRACT The placenta plays a crucial role during pregnancy ensuring fetal growth and development. Successful pregnancy hinges upon placental adaptations to the maternal environment. The human placenta is classified as hemochorial and is characterized by extensive intrauterine trophoblast cell invasion. During the course of the gestation, invasive trophoblast cells or extravillous trophoblast, as they are referred to in humans, migrate from the placenta into the uterine parenchyma where they act to anchor the placenta to the uterus and remodel uterine spiral arteries. Uterine vascular remodeling is central to providing adequate nutrient flow to the fetus and normal fetal development. Abnormalities in the organization and structure of trophoblast cell populations are associated with pregnancy disease states. Shallow trophoblast invasion and aberrant modification of the uterine vasculature are directly linked to preeclampsia, early pregnancy loss, intrauterine growth restriction, pre-term birth, and placental abruption. On the other hand, excessive trophoblast invasion, as in disorders such as placenta accreta, jeopardizes pregnancy, maternal health, and future-child bearing. Molecular events that trigger the differentiation of invasive trophoblast cell lineages and trophoblast-guided uterine spiral artery remodeling are poorly understood. Experimental tools are in-hand to identify candidate regulators of the invasive trophoblast cell lineage and to test their physiological relevance. Placentation in the rat is characterized by extensive intrauterine trophoblast invasion and trophoblast-guided spiral artery remodeling resembling developmental processes evident in human placentation. Global genome-editing in the rat is an effective tool for investigating a sub-set of candidate genes implicated in regulating deep placentation but not all candidate genes regulating deep placentation. Some genes possess multiple roles in embryogenesis, precluding an evaluation of a specific role in the invasive trophoblast cell lineage. The generation of a conditional allele circumvents this issue. In the proposed research, we will generate a rat model for generating invasive trophoblast-lineage specific conditional alleles using Crispr/Cas9 genome editing. In this project, we generate a transgenic rat model expressing Cre recombinase in invasive trophoblast cells under control of Prl7b1 regulatory sequences and validate the Prl7b1- Cre driver rat strain. The proposed experiments will provide valuable new tools for the scientific community to discern roles for candidate genes in the regulation of trophoblast invasion and trophoblast-guided uterine spiral artery remodeling, and thus a novel experimental paradigm to explore the etiology of high-risk pregnancies.