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.

项目摘要 胎盘是平衡母体营养供应的调节和交换屏障 与怀孕期间胎儿的代谢需求有关。胎盘发育不足， 功能障碍导致一系列不利的胎儿和母体结果。前三个月是一个关键时期， 建立适当的胎盘，但我们目前的知识，第一个三个月的发展， 不充分，对整个怀孕期间胎盘发育和功能的理解受到阻碍， 缺乏纵向样品的获取途径和缺乏合适的体外模型系统。 在胎盘形成早期，滋养层细胞，一种专门的胎盘干细胞，分化成两种类型： 绒毛外滋养层细胞（EVT）侵入母体螺旋动脉，将胎盘锚在胎盘中。 蜕膜，并且对于形成功能齐全的胎盘的强大血管基础至关重要。的 合胞体滋养层（SYN）是一种多核上皮细胞，充当母胎交换表面。尽管 它们的重要性，了解滋养层细胞谱系特化和分化的调节， 由于缺乏合适的细胞模型系统和从早期就能接触胎盘组织， 怀孕类器官是自组织和繁殖的三维（3D）培养模型系统， 来源于干细胞。它们可以通过操纵生长来引导离体生长到微型器官结构中 条件和提供驱动表型特异性细胞发育的发育线索。最近，一名 妊娠期人类滋养层类器官系统已经开发出来。然而，胎盘是胎儿组织， 导致与研究中使用终止样本相关的伦理问题。非人灵长类 (NHP)为这个问题提供了一个解决方案。我们建议生成妊娠早期NHP类器官模型，并测试 通过超声引导下绒毛膜绒毛活检获取妊娠早期胎盘样本的可行性 采样（CVS），从而避免了终止的需要。使用类器官进行体外培养的第二个挑战 实验是细胞极性。具体地，基质悬浮液的使用通常使顶端细胞表面取向为 该中心产生了一种“由内而外”的类器官结构，从而限制了它们在胎盘屏障研究中的应用。 重要的是，其他类器官系统的最新进展表明， 将3D结构转化为2D细胞层。 该提案的首要前提是开发一种新的体外工具，以扩展我们对以下内容的理解： 转化动物模型中的早期胎盘发育和功能。在科学目标范围内，我们将 除了收集整个胎盘组织用于类器官制备外，还利用CVS获得胎盘活检 直接比较两种抽样方法。类器官将被诱导分化， 改变细胞极性的条件。这项新颖的工作将建立一个管道，以前无法进入 研究正常和紊乱的胎盘功能的途径。