Developing a novel approach to the nonhuman primate placental organoid model

开发非人灵长类胎盘类器官模型的新方法

• 批准号: 10589154
• 负责人: Victoria HJ Roberts
• 金额: $ 26.25万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-10 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589154
• 关键词: 3-Dimensional; Animal Model; Apical; Biological Models; Biopsy; Biopsy Specimen; Blood Vessels; Cell Communication; Cell Differentiation Induction; Cell Differentiation process; Cell Lineage; Cell Polarity; Cell model; Cell surface; Cells; Characteristics; Chorionic Villi Sampling; Clinical; Cues; Decidua; Development; Digestion; Discipline of obstetrics; Epithelium; Equilibrium; Ethics; Extracellular Matrix; Fetal Tissues; First Pregnancy Trimester; Foundations; Functional disorder; Growth; Histologic; Human; In Vitro; Invaded; Investigation; Knowledge; Macaca; Macaca mulatta; Maternal-Fetal Exchange; Mediator; Metabolic; Methodology; Methods; Modeling; Nutritional; Organ; Organoids; Pathway interactions; Phenotype; Placenta; Placentation; Pregnancy; Prenatal Diagnosis; Preparation; Procedures; Process; Property; Protocols documentation; Regulation; Research; Resources; Sampling; Source; Specific qualifier value; Spiral Artery of the Endometrium; Structure; Surface; Suspensions; Syncytiotrophoblast; System; Techniques; Termination of pregnancy; Testing; Time; Tissue Banks; Tissue Model; Tissue Sample; Tissues; Villous; Work; avoid pregnancy; cell dimension; cell type; critical period; experimental study; feasibility testing; fetal; in vitro Assay; in vitro Model; insight; maternal outcome; minimally invasive; nonhuman primate; novel; novel strategies; placental stem cell; pregnant; sample collection; self organization; stem cells; three dimensional structure; tissue culture; tool; trophoblast; trophoblast stem cell; two-dimensional; ultrasound

PROJECT SUMMARY The placenta is the regulatory and exchange barrier that functions to balance maternal nutritional supply with fetal metabolic demands during pregnancy. Inadequate placental development and subsequent dysfunction results in a range of adverse fetal and maternal outcomes. The first trimester is a crucial time for the establishment of appropriate placentation, yet our current knowledge of first trimester development is inadequate, and understanding of placental development and function throughout pregnancy is impeded by the lack of access to longitudinal samples and a lack of suitable in vitro model systems. In early placental formation, trophoblast cells, a specialized placental stem cell, differentiate into two types: extravillous trophoblast cells (EVTs), which invade the maternal spiral arteries, anchor the placenta in the decidua and are critical for forming a strong vascular foundation for a fully functioning placenta. The syncytiotrophoblast (SYN), a multinucleated epithelium, serves as the maternal-fetal exchange surface. Despite their importance, understanding the regulation of trophoblast cell lineage specification and differentiation has been hindered by the lack of appropriate cellular model systems and access to placental tissue from early gestation. Organoids are self-organizing and propagating 3-dimensional (3D) culture model systems that are derived from stem cells. They can be directed to grow ex vivo in to mini organ structures by manipulating growth conditions and providing developmental cues that drive phenotype-specific cell development. Recently, a first trimester human trophoblast organoid system has been developed. However, the placenta is a fetal tissue which leads to ethical concerns associated with the use of termination samples in research. The nonhuman primate (NHP) offers a solution to this problem. We propose to generate a first trimester NHP organoid model, and test the feasibility of obtaining first trimester placenta samples through the use of ultrasound-guided chorionic villous sampling (CVS), thus avoiding the need for termination. A second challenge to the use of organoids for in vitro experiments is cell polarity. Specifically, use of a matrix suspension typically orientates the apical cell surface to the center creating an `inside out' organoid structure, thus limiting their utility in placental barrier studies. Importantly, recent advances in other organoid systems have demonstrated the ability to alter the composition of the extracellular matrix to convert 3D structures to 2D cell layers. The overarching premise of this proposal is to develop a new ex vivo tool to expand our understanding of early placental development and function in a translational animal model. Within the scientific objectives we will utilize CVS to obtain placental biopsies, in addition to whole placental tissue collection for organoid preparations to directly compare the two sampling methodologies. Organoids will be induced to differentiate, and culture conditions manipulated to alter cell polarity. This novel work will establish a pipeline for previously inaccessible pathways for the study of normal and perturbed placental function.

项目总结 胎盘是调节和交换屏障，起到平衡母体营养供应的作用。 在怀孕期间有胎儿新陈代谢需求。胎盘发育不全和随后 功能障碍会导致一系列不良的胎儿和产妇结局。前三个月是一个关键的时期 建立适当的胎盘，但我们目前对早期妊娠发育的了解是 不充分，对胎盘发育和功能的理解在整个怀孕过程中受到阻碍 缺乏获得纵向样本的途径，以及缺乏合适的体外模型系统。 在胎盘形成的早期，滋养层细胞，一种专门的胎盘干细胞，分化为两种类型： 绒毛外滋养层细胞(EVT)侵袭母体螺旋动脉，将胎盘固定在 蜕膜，对形成充分发挥功能的胎盘的强大血管基础至关重要。这个 合体滋养层细胞(SYN)是一种多核上皮细胞，是母胎之间的交换面。尽管 它们的重要性，了解滋养层细胞谱系的规范和分化的调节 由于缺乏适当的细胞模型系统和早期获取胎盘组织的途径而受到阻碍 怀孕了。有机体是自组织和传播三维(3D)培养模型系统， 来源于干细胞。它们可以通过控制生长，在体外生长到微型器官结构中。 并提供驱动特定表型细胞发育的发育线索。最近，第一次 已研制出妊娠三个月的人滋养层类器官系统。然而，胎盘是一种胎儿组织， 导致与在研究中使用终止样本相关的伦理问题。非人灵长类 (NHP)为这一问题提供了解决方案。我们建议生成一个怀孕早期的NHP有机物模型，并进行测试 超声引导绒毛采集早孕胎盘标本的可行性研究 采样(CVS)，从而避免了终止的需要。在体外使用有机化合物面临的第二个挑战 实验是细胞的极性。具体地说，使用基质悬浮液通常将顶端细胞表面定向到 该中心创造了一种‘内向外’的器官结构，从而限制了它们在胎盘屏障研究中的应用。 重要的是，其他有机系统的最新进展已经证明了改变组成的能力。 以将3D结构转换为2D细胞层。 这项提议的首要前提是开发一种新的体外工具来扩大我们对 翻译动物模型中胎盘的早期发育和功能。在科学目标范围内，我们将 利用CVS获取胎盘活检，并收集整个胎盘组织用于有机制剂 直接比较两种抽样方法。有机化合物将被诱导分化，并培养 被操控以改变细胞极性的条件。这项新颖的工作将建立一条管道，将以前无法进入的 研究正常和紊乱胎盘功能的途径。