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/V005367/1
• 负责人: Naomi Moris
• 金额: $ 80.84万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV005367%2F1
• 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 块单独的骨头组成，呈线性排列，呈连续结构，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质，并且起源于体细胞发生过程，体细胞发生过程发生在早期胚胎中，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（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，其严重程度多种多样，在极端情况下，可能会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至关重要。体节发生的过程已经在许多不同的模型生物体中进行了研究，包括鱼、鸡和小鼠。遗传分析表明，中胚层的周期性分裂与某些基因的表达在空间和时间上的振荡有关。这些基因的突变会导致各种分割缺陷，表现为脊柱和胸肌组织的畸变，如脊柱缺陷患者中所见。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制关系。对人类这些基因的分析因在分割阶段难以获取胚胎而受到阻碍。过去几年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，可以使用它们来模拟与小鼠和人类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，无法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。该项目旨在开发一种新的模型系统来在体外研究正常和异常的人类体节发生。这将通过将两个具有互补专业知识的团队聚集在一起来实现。位于京都的一家公司已经建立了人类 PSC 的体细胞发生模型，该模型以无序的方式重现了骨骼和肌肉的振荡和分化。位于剑桥的第二个小组首创了“类原肠胚”，这是一种基于三维 PSC 的早期哺乳动物发育模型，最近已扩展到人类 PSC。原肠胚以前后极性概括了体节发生的基本空间和时间方面。该项目将把两个小组聚集在一起，优化人类原肠胚，用于研究体节的形成和分化，并测试一些来自椎骨缺陷患者的 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学，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

In vitro teratogenicity testing using a 3D, embryo-like gastruloid system.

• DOI: 10.1016/j.reprotox.2021.08.003
• 发表时间: 2021-10
• 期刊: Reproductive toxicology (Elmsford, N.Y.)
• 作者: Mantziou V;Baillie-Benson P;Jaklin M;Kustermann S;Arias AM;Moris N
• 通讯作者: Moris N