3D Human Axial Development In Vitro: using novel human in vitro somitogenesis models to study birth defects with patient-relevant iPS cell lines

3D 人体轴体外发育：使用新型人体体外体细胞发生模型研究患者相关 iPS 细胞系的出生缺陷

• 批准号: MR/V005367/2
• 负责人: Naomi Moris
• 金额: $ 69.17万
• 依托单位: The Francis Crick Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV005367%2F2
• 关键词: 3D; Human; Axial; Development; Vitro

The vertebral column is a key element of the human body, made up of 33 individual bones arranged linearly in a continuous structure that houses the spinal cord and provides support for the ribs and associated muscles. This structure reveals the segmental nature of our body and has its origin in the process of somitogenesis, an event that takes place in the early embryo where elongation of the body axis leads to the sequential partitioning of a group of cells, the mesoderm, into discrete blocks, called somites. These somites will give rise to progenitors for bone and muscle derivatives, including the vertebrae. Structural abnormalities in vertebrae formation are a common pathology of early development, and span a range of severities that, in extreme forms, can have dramatic effects on the life of individuals. It is therefore critically important that we better understanding the basis of these disorders.The process of somitogenesis has been studied in many different model organisms, including fish, chicken and mice. Genetic analysis has revealed that the periodic partitioning of the mesoderm is associated with oscillations in the expression of certain genes in space and time. Mutations in these genes lead to a variety of segmentation defects that manifest as aberrations in the organisation of the vertebral column and thoracic muscles, as seen in patients with vertebral column defects. However, there are differences in the development of mice and humans that make it difficult to infer causal, mechanistic relationships between mutations and specific syndromes.Analysis of these genes in humans is hampered by the difficulty of accessing embryos at the stages of segmentation. One approach to this problem that has emerged over the last few years is Pluripotent Stem Cells (PSCs), which have been shown to recapitulate many developmental events in a laboratory setting. In particular, it is possible to use them to model gene expression patterns associated with somitogenesis in mouse and human. However, these models lack the spatial organization of the embryo that subdivides the length of the mesoderm into constantly proportioned territories and leads to the development of somite structures. This project aims to develop a new model system to study normal and aberrant human somitogenesis in vitro. This will be achieved by bringing together two groups with complementary expertise. One, based in Kyoto, has established a somitogenesis model from human PSCs that recapitulates oscillations and differentiation into bone and muscle, but in a disorganized manner. A second group, based in Cambridge, has pioneered 'gastruloids', a three-dimensional PSC-based model of early mammalian development recently extended to human PSCs. Gastruloids recapitulate the basic spatial and temporal aspects of somitogenesis in 3D with an anteroposterior polarity. The project will bring the two groups together to optimize humans gastruloids for the study of the formation and differentiation of somites, and to test a number of iPSC lines derived from patients with vertebral defects. The outcome of the project will be the establishment of a new platform for the detection and analysis of pathologies associated with the segmentation of the mesoderm in human embryos, leading to a better understanding of the causes of these syndromes.

椎骨是人体的关键要素，由33个单独的骨头组成，该骨骼在连续结构中线性排列，该结构容纳脊髓，并为肋骨和相关的肌肉提供了支撑。这种结构揭示了我们身体的分段性质，并起源于体内生成过程，这一事件发生在早期的胚胎中，在早期的胚胎中，人体轴的伸长导致一组细胞（中胚层）的顺序分配到离散的块中，称为somites。这些节点将引起骨和肌肉衍生物（包括椎骨）的祖细胞。椎骨形成的结构异常是早期发育的常见病理，并且跨越了一系列严重性，这些严重性以极端形式可以对个人的生活产生巨大影响。因此，至关重要的是，我们可以更好地理解这些疾病的基础。在许多不同的模型生物（包括鱼类，鸡肉和小鼠）中已经研究了体型的过程。遗传分析表明，中胚层的周期性分配与时空中某些基因表达的振荡有关。这些基因的突变导致各种分割缺陷，这些缺陷表现为椎骨和胸肌的组织中的畸变，如椎骨缺陷患者所见。然而，小鼠和人类的发展存在差异，使得很难推断出突变与特定综合症之间的机理关系。这些基因在人类中的分析受到了在分段阶段获得胚胎的困难。在过去几年中，解决此问题的一种方法是多能干细胞（PSC），这些干细胞已被证明可以在实验室环境中概括许多发展事件。特别是，可以使用它们来建模与小鼠和人类中的体体发生相关的基因表达模式。但是，这些模型缺乏胚胎的空间组织，该胚胎将中胚层的长度细分为不断成比例的领土，并导致体系结构的发展。该项目旨在开发一种新的模型系统，以研究体外人类的正常和异常的人体生成。这将通过将两个互补专业知识的小组汇集在一起​​来实现。一家位于京都的人已经从人类PSC建立了一个体体发生模型，该模型概括了振荡并分化为骨骼和肌肉，但以混乱的方式。一组位于剑桥的第二组率先启用了“胃肠道”，这是一种基于三维PSC的早期哺乳动物发育模型，该模型最近扩展到人类PSC。胃突对3D的基本空间和时间生成概括了前后极性的基本空间和时间方面。该项目将将这两个小组聚集在一起，以优化人类的胃肠样，以研究体系的形成和分化，并测试许多来自椎骨缺陷患者的IPSC线。该项目的结果将是建立一个新平台，用于检测和分析与人类胚胎中中胚层相关的病理学，从而更好地理解了这些综合征的原因。

项目成果

Reconstituting human somitogenesis in vitro

体外重建人类体节发生

• DOI: 10.1038/s41586-022-05649-2
• 发表时间: 2022
• 期刊: Nature
• 影响因子: 64.8
• 作者: Yamanaka Yoshihiro;Hamidi Sofiane;Yoshioka-Kobayashi Kumiko;Munira Sirajam;Sunadome Kazunori;Zhang Yi;Kurokawa Yuzuru;Ericsson Rolf;Mieda Ai;Thompson Jamie L.;Kerwin Janet;Lisgo Steven;Yamamoto Takuya;Moris Naomi;Martinez-Arias Alfonso;Tsujimura Taro;Alev
• 通讯作者: Alev