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: 10371545
• 负责人: Victoria HJ Roberts
• 金额: $ 21.88万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371545
• 关键词: 3-Dimensional; Animal Model; Apical; Biological Models; Biopsy; Biopsy Specimen; Blood Vessels; Cell Communication; Cell Differentiation process; Cell Lineage; Cell Polarity; Cell model; Cell surface; Cells; Characteristics; Chorionic Villi Sampling; Clinical; Cues; Decidua; Development; Digestion; Dimensions; Discipline of obstetrics; Epithelial; 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 of activation protein; 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; 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; 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; stem cell differentiation; 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.

项目概要 胎盘是调节和交换屏障，具有平衡母体营养供应的作用 怀孕期间胎儿的代谢需求。胎盘发育不足及后续 功能障碍会导致一系列不良的胎儿和母亲结局。孕早期是关键时期 建立适当的胎盘，但我们目前对妊娠早期发育的了解是 不足，并且对整个怀孕期间胎盘发育和功能的了解受到阻碍 缺乏纵向样本和合适的体外模型系统。 在胎盘形成的早期，滋养层细胞（一种特殊的胎盘干细胞）分化为两种类型： 绒毛外滋养层细胞（EVT）侵入母体螺旋动脉，将胎盘固定在胎盘中 蜕膜对于形成功能齐全的胎盘的强大血管基础至关重要。这 合体滋养层（SYN）是一种多核上皮，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规范和分化的调节已经 由于缺乏适当的细胞模型系统和早期胎盘组织的获取而受到阻碍 妊娠。类器官是自组织和传播 3 维 (3D) 培养模型系统， 源自干细胞。通过操纵生长，可以引导它们离体生长成微型器官结构 条件并提供驱动表型特异性细胞发育的发育线索。近日，第一 已开发出妊娠期人类滋养层类器官系统。然而，胎盘是胎儿的组织， 导致与在研究中使用终止样本相关的伦理问题。非人类灵长类动物 （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，并进行测试 通过超声引导绒毛膜绒毛获取早孕期胎盘样本的可行性 采样（CVS），从而避免了终止的需要。体外使用类器官的第二个挑战 实验是细胞极性。具体来说，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“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效用。 重要的是，其他类器官系统的最新进展已经证明了改变其组成的能力 细胞外基质将 3D 结构转换为 2D 细胞层。 该提案的首要前提是开发一种新的体外工具来扩大我们对 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 获取胎盘活检 直接比较两种抽样方法。类器官将被诱导分化，并进行培养 操纵条件来改变细胞极性。这项新颖的工作将为以前无法​​访问的项目建立一条管道 研究正常和异常胎盘功能的途径。