A new animal model for investigating the invasive trophoblast lineage

用于研究侵袭性滋养层谱系的新动物模型

• 批准号: 10448294
• 负责人: Khursheed Iqbal
• 金额: $ 23.9万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-10 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10448294
• 关键词: Abruptio Placentae; Alleles; Animal Model; CRISPR/Cas technology; Candidate Disease Gene; Cell Lineage; Cells; Communities; Cre driver; Deposition; Development; Developmental Process; Disease; Early Diagnosis; Embryonic Development; Ensure; Enterobacteria phage P1 Cre recombinase; Environment; Etiology; Evaluation; Event; Failure; Fetal Development; Fetal Growth; Fetal Growth Retardation; Fetus; Functional disorder; Future; Generations; Genes; Growth and Development function; Hand; Health; Hemochorial Placental Development; High-Risk Pregnancy; Human; In Situ Hybridization; Invaded; Knock-in; Labyrinth; Link; Maternal Health; Missouri; Modeling; Modification; Molecular; Mothers; Nutrient; Physiological; Placenta; Placenta Accreta; Placentation; Play; Population; Positioning Attribute; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Premature Birth; Property; Rat Strains; Rattus; Regulation; Research; Resources; Role; Spiral Artery of the Endometrium; Testing; Therapeutic Intervention; Tissues; Transgenic Organisms; United States National Institutes of Health; Universities; Uterus; Vascular remodeling; child bearing; early pregnancy loss; experimental study; fetal; genome editing; healthy pregnancy; in vivo; novel; organizational structure; postnatal; single-cell RNA sequencing; targeted treatment; tool; trafficking; trophoblast; trophoblast stem cell

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.

项目摘要/摘要 胎盘在怀孕期间起着至关重要的作用，确保胎儿生长和发育。成功的 怀孕对孕产妇环境的适应性呈现。人胎盘被归类为 血小板和以宫内滋养细胞侵袭的广泛特征。在 妊娠，侵入性滋养细胞细胞或跨性滋养细胞，如人类所指的那样，从 胎盘进入子宫实质，在那里它们作用将胎盘锚定在子宫和改造子宫 螺旋动脉。子宫血管重塑对于提供足够的营养流向胎儿和正常 胎儿发育。滋养细胞细胞种群的组织和结构异常 与怀孕疾病状态。子宫脉管系统的浅滋养细胞入侵和异常修饰 直接与先兆子痫，早期妊娠丧失，宫内生长限制，预期出生和 胎盘破裂。另一方面，过多的滋养细胞入侵，例如胎盘cocta等疾病， 危害怀孕，孕产妇的健康和未来的孩子。触发分化的分子事件 侵入性滋养细胞细胞谱系和滋养细胞引导的子宫螺旋动脉的重塑很差 理解。实验工具是识别侵入性滋养细胞细胞的候选调节因子 谱系并测试其生理相关性。大鼠胎盘的特征是宫内广泛 滋养细胞的入侵和滋养细胞引导的螺旋动脉重塑类似于发育过程 在人类胎盘中很明显。大鼠中的全球基因组编辑是调查子集的有效工具 候选基因涉及调节深滤料，但并非所有调节深层的候选基因 胎盘。某些基因在胚胎发生中具有多种作用，排除了对特定作用的评估 在侵入性滋养细胞谱系中。有条件的等位基因的产生绕过了这个问题。在 拟议的研究，我们将生成一个大鼠模型，以生成侵入性滋养细胞固定特定条件 使用CRISPR/CAS9基因组编辑的等位基因。在这个项目中，我们生成一个表达Cre的转基因大鼠模型 在控制PRL7B1调节序列的侵入性滋养细胞细胞中的重组酶，并验证PRL7B1-- CRE驱动老鼠应变。拟议的实验将为科学界提供宝贵的新工具 候选基因在调节滋养细胞入侵和滋养细胞引导的子宫螺旋中的识别作用 动脉重塑，因此是一种新型的实验范式，以探索高危妊娠的病因。