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.

脊柱是人体的关键元素，由33个单独的骨骼组成，这些骨骼以连续的结构线性排列，容纳脊髓并为肋骨和相关肌肉提供支撑。这种结构揭示了我们身体的节段性，并起源于体节发生的过程，这是发生在早期胚胎中的一个事件，在该事件中，体轴的伸长导致一组细胞（中胚层）顺序划分成离散的块，称为体节。这些体节将产生骨骼和肌肉衍生物的祖细胞，包括椎骨。椎骨形成中的结构异常是早期发育的常见病理学，并且跨越一系列严重程度，在极端形式中，可以对个体的生活产生巨大影响。因此，我们更好地了解这些疾病的基础是至关重要的。已经在许多不同的模式生物中研究了体节发生的过程，包括鱼、鸡和小鼠。遗传分析表明，中胚层的周期性分配与某些基因在空间和时间上的表达振荡有关。这些基因的突变会导致各种节段缺陷，表现为脊柱和胸肌组织的畸变，如脊柱缺陷患者所见。然而，小鼠和人类的发育存在差异，这使得很难推断突变和特定综合征之间的因果关系和机械关系。在分割阶段难以进入胚胎，阻碍了对人类这些基因的分析。在过去几年中出现的解决这个问题的一种方法是多能干细胞（PSC），它已被证明可以在实验室环境中重现许多发育事件。特别是，有可能使用它们来模拟与小鼠和人类中的体节发生相关的基因表达模式。然而，这些模型缺乏胚胎的空间组织，即将中胚层的长度细分为恒定比例的区域，并导致体节结构的发育。本研究旨在建立一种新的体外研究正常和异常人类体节发生的模型系统。这将通过将具有互补专长的两个小组聚集在一起来实现。其中一个位于京都，已经建立了一个来自人类PSC的体节发生模型，该模型重现了骨骼和肌肉的振荡和分化，但以一种混乱的方式。位于剑桥的第二个研究小组开创了“类胃体”，这是一种基于PSC的早期哺乳动物发育三维模型，最近扩展到人类PSC。Gastruloids在3D中概括了体节发生的基本空间和时间方面，具有前后极性。该项目将把两个小组聚集在一起，优化人类类胃体，用于研究体节的形成和分化，并测试一些来自脊椎缺陷患者的iPSC系。该项目的成果将是建立一个新的平台，用于检测和分析与人类胚胎中胚层分割相关的病理，从而更好地了解这些综合征的原因。

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