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. !

项目摘要/摘要 在动态组织尺度的背景下，精确的细胞命运规范是如何进行的 重排是发育生物学中最令人兴奋的问题之一。在 我们旨在破译命运收购之间的相互作用， 在 人神经。我们最近开发了一个强大的协议，该协议允许 为了产生极其可重现的人类神经素：自组织 概括组织的人类胚胎干细胞（HESC）组件 通过组织神经神经的神经阶段的外胚层室 在粘合剂上同一菌落中的波峰，位置和表皮种群 微图案。这种自组织非常可复制，可以是 用亚细胞分辨率和数百个菌落实时量化。 在这项新型技术中，我们提出了三个具体目标。首先是 揭示细胞 - 细胞信号传导驱动自组织的机制。第二个 是通过现场将信号与命运的获取和形态发生结合 报告基因成像和依赖时间的单细胞RNASEQ。最后，第三个目标 着重于外胚层的原点和亚群的全部表征 衍生物及其体内验证通过与 阶段匹配的果酱胎儿样品和小鸡的嫁接实验 胚胎。大量同质的人神经素的产生， 在一段时间内可以动态遵循外胚层命运的自组织 一周，具有亚细胞分辨率，不仅解决了固有的 在大脑器官中观察到的异质性，但为我们提供了独特的 在人类胚胎模型中研究这些事件的机会。这将有一个 在基础研究和临床应用中都有很大的影响 已经在地平线上了。 呢