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.

项目总结/摘要 胎盘在怀孕期间起着至关重要的作用，确保胎儿的生长和发育。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 血绒膜的，其特征在于广泛的子宫内滋养层细胞浸润。过程中 妊娠期间，侵袭性滋养层细胞或绒毛外滋养层（在人类中被称为滋养层）从 胎盘进入子宫实质，在那里它们起作用以将胎盘锚在子宫上并重塑子宫 螺旋动脉子宫血管重塑对于为胎儿和正常人提供充足的营养流至关重要 胎儿发育滋养层细胞群的组织和结构异常与 妊娠疾病的国家。滋养细胞浅侵及子宫血管的异常改变 与先兆子痫、早孕流产、宫内生长受限、早产直接相关， 胎盘萎缩另一方面，过度的滋养层侵入，如胎盘植入， 危害怀孕、产妇健康和未来生育。引发分化的分子事件 浸润性滋养层细胞谱系和滋养层引导的子宫螺旋动脉重塑的发生率很低， 明白实验工具是在手，以确定候选调节浸润性滋养层细胞 血统，并测试其生理相关性。大鼠胎盘形成的特点是广泛的子宫内 滋养层浸润和滋养层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局基因组编辑是研究基因组亚组的有效工具。 参与调节深层胎盘形成的候选基因，但不是所有的候选基因都调节深层胎盘形成。 胎座式有些基因在胚胎发生中具有多重作用，因此无法对某一特定作用进行评价 在侵袭性滋养层细胞谱系中。条件等位基因的产生避免了这个问题。在 建议的研究，我们将产生一个大鼠模型，用于产生侵袭性滋养层细胞系特异性条件 使用Crispr/Cas9基因组编辑的等位基因。在本项目中，我们建立了表达Cre的转基因大鼠模型， 在Prl 7 b1调控序列的控制下，在侵袭性滋养层细胞中重组酶，并验证Prl 7 b1- Cre驱动大鼠品系。拟议的实验将为科学界提供宝贵的新工具， 识别候选基因在滋养细胞侵袭和滋养细胞引导的子宫螺旋调节中的作用 动脉重塑，从而一种新的实验范式，探索高危妊娠的病因。