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 胎盘是平衡孕产妇营养供应的调节和交换障碍 怀孕期间的胎儿代谢需求。胎盘发育不足，随后 功能障碍会导致一系列不良的胎儿和产妇结局。头三个月是至关重要的时间 建立适当的胎盘，但是我们目前对孕期发展的了解是 不足，对整个怀孕期间胎盘发育和功能的理解受到了阻碍 缺乏获得纵向样品的访问权限，缺乏适合的体外模型系统。 在早期胎盘形成中，滋养细胞（一种专门的胎盘干细胞）分为两种类型： 侵入母体螺旋动脉的额外滋养细胞细胞（EVT）将胎盘固定在 Decidua，对于为胎盘功能齐全的良好的血管基础至关重要。这 多核上皮的合成细胞塑性细胞（SYN）作为母亲交换表面。尽管 它们的重要性，了解滋养细胞细胞谱系规范的调节和分化的调节 由于缺乏适当的蜂窝模型系统和从早期进入胎盘组织而受到阻碍 妊娠。器官是自组织和繁殖的3维（3D）培养模型系统 源自干细胞。可以通过操纵生长来指示它们在小体内生长到迷你器官结构 条件并提供推动表型特异性细胞发育的发育线索。最近，第一个 已经开发了三个月的人滋养细胞器官系统。但是，胎盘是一种胎儿组织 导致与在研究中使用终止样本有关的道德问题。非人类的灵长类动物 （NHP）为此问题提供了解决方案。我们建议生成第一三个月的NHP器官模型，然后测试 通过使用超声引导的绒毛膜绒毛获得先发孕期样品的可行性 采样（CVS），因此避免了终止的需求。对使用器官进行体外的第二个挑战 实验是细胞极性。具体而言，使用矩阵悬浮液通常将顶端细胞表面定向 该中心产生了“内部”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证明了改变组成的能力 细胞外基质将3D结构转换为2D细胞层。 该提案的总体前提是开发一种新的离体工具，以扩展我们对 转化动物模型中的早期胎盘发育和功能。在科学目标中，我们将 除了整个胎盘组织收集外，还利用CV获得胎盘活检 直接比较两种采样方法。器官将被诱导分化和培养 操纵以改变细胞极性的条件。这项新颖的工作将建立以前无法访问的管道 研究正常和干扰胎盘功能的途径。