Micropattern differentiation and morphogenesis of the human ectoderm

人类外胚层的微模式分化和形态发生

• 批准号: 10665577
• 负责人: ALI H BRIVANLOU
• 金额: $ 35.71万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-24 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665577
• 关键词: Adhesives; Automobile Driving; Basic Science; Biological Assay; CRISPR/Cas technology; Callithrix; Cells; Cellular biology; Central Nervous System; Cerebrum; Chick Embryo; Clinic; Coin; Development; Developmental Biology; Disease; Dissection; Ectoderm; Ectoderm Cell; Embryo; Embryonic Development; Environment; Epidermis; Event; Fibroblast Growth Factor; Generations; Genes; Germ Layers; Heterogeneity; Human; Image; In Vitro; Knowledge; Link; Mediating; Modeling; Molecular; Molecular Profiling; Morphogenesis; Neural Crest; Neurula; Organoids; Pathologic Processes; Pattern; Phase; Population; Positioning Attribute; Process; Protocols documentation; Reporter; Reproducibility; Resolution; Sampling; Side; Signal Transduction; Specific qualifier value; Specificity; Stains; Standardization; Structure; Time; Tissues; Validation; cell assembly; cell fate specification; cell type; clinical application; combinatorial; data integration; direct application; experimental study; fetal; human embryonic stem cell; human fetus tissue; human model; in vivo; intercellular communication; mathematical model; neural; new technology; primate development; progenitor; self organization; single-cell RNA sequencing; spatiotemporal

Project Summary/Abstract How accurate cell fate specification occurs in the context of dynamic tissue-scale rearrangements is one of the most exciting questions in developmental biology. In this proposal we aim at deciphering the interplay between fate acquisition, patterning and morphogenesis of the ectodermal germ layer in the context of human neurulation. We have recently developed a robust protocol which allows for the generation of extremely reproducible human neuruloids: self-organized human Embryonic Stem Cell (hESC) assemblies that recapitulate the organization of the ectodermal compartment at neurulation stages by organizing neural, neural crest, placodes and epidermis populations within the same colony on adhesive micropatterns. This self-organization is extremely reproducible and can be quantified with sub-cellular resolution and in real time over hundreds of colonies. Armed with this novel technology, we propose three specific aims. The first is to unravel the mechanism of cell-cell signaling driving self-organization. The second is to integrate signaling with fate acquisition and morphogenesis through live reporter imaging and time dependent single cell RNAseq. Finally, the third aim focuses on the full characterization of the origin and sub-populations of ectodermal derivatives and their in vivo validation by performing side by side comparisons with stage-matched marmoset fetal samples and grafting experiments in chick embryos. The generation of large numbers of homogenous human neuruloids, where self-organization of ectodermal fate can be followed dynamically for a period of one week, with sub-cellular resolution, not only solves the inherent heterogeneity observed in cerebral organoids, but provides us a unique opportunity to study these events in models of human embryos. This will have a high impact in both basic research as well as clinical application, a prospect already on the horizon. !

项目概要/摘要 在动态组织规模的背景下如何准确地进行细胞命运规范 重排是发育生物学中最令人兴奋的问题之一。在 这个提案我们的目的是破译命运获取之间的相互作用， 外胚层胚层的模式和形态发生 人类神经系统。我们最近开发了一个强大的协议，允许 用于生成极其可重复的人类神经样细胞：自组织 人类胚胎干细胞 (hESC) 组装体概括了该组织 通过组织神经，神经形成阶段的外胚层室 嵴、基板和表皮群体位于粘合剂上的同一群体内 微图案。这种自组织具有极强的可重复性，并且可以 以亚细胞分辨率对数百个菌落进行实时量化。 借助这项新技术，我们提出了三个具体目标。第一个是 揭示细胞间信号传导驱动自组织的机制。第二个 是通过活体将信号传导与命运获取和形态发生整合起来 报告成像和时间依赖性单细胞 RNAseq。最后，第三个目标 侧重于外胚层起源和亚群的全面表征 衍生物及其体内验证通过并排比较 阶段匹配的狨猴胎儿样本和雏鸡移植实验 胚胎。大量同质人类神经样细胞的产生， 外胚层命运的自组织可以在一段时间内动态跟踪 一周的时间，具有亚细胞分辨率，不仅解决了固有的问题 在大脑类器官中观察到的异质性，但为我们提供了独特的 有机会在人类胚胎模型中研究这些事件。这将有一个 在基础研究和临床应用方面均具有较高影响力，前景广阔 已经在地平线上了。 ！