Deconstructing human body plan development with stem cells

用干细胞解构人体计划制定

• 批准号: 10644147
• 负责人: Yuchuan Miao
• 金额: $ 11.56万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644147
• 关键词: 3-Dimensional; Adopted; Animal Model; Anterior; Biological; Capsicum; Cell Differentiation process; Cell Maturation; Cell Separation; Cells; Complex; Data; Development; Development Plans; Developmental Process; Digit structure; Disease; Distal; Embryo; Engineering; Ensure; Epithelium; Ethics; Etiology; Foundations; Future; Genes; Genetic; Goals; Human; Human Development; Human body; In Vitro; Individual; Investigation; Knowledge; Length; Light; Limb Bud; Limb Development; Limb structure; Link; Literature; Mediating; Mentors; Mesenchymal; Mesoderm Cell; Modeling; Molecular; Morphogenesis; Musculoskeletal Diseases; Organ; Organoids; Paraxial Mesoderm; Pattern; Pattern Formation; Periodicals; Peripheral Nervous System; Phase; Pluripotent Stem Cells; Postdoctoral Fellow; Proxy; Regenerative Medicine; Research; Resolution; Role; Segmentation Clock Pathway; Sodium Chloride; Somites; Specific qualifier value; Structure; Testing; Tissues; Training; Work; appendage; blastomere structure; career; differentiation protocol; embryo cell; experimental study; human model; human pluripotent stem cell; in vitro Model; in vivo; induced pluripotent stem cell; insight; medical specialties; novel; progenitor; self organization; somitogenesis; spatiotemporal; spine bone structure; stem cells; three dimensional cell culture; three-dimensional modeling

How the precise patterns are achieved during development of the human body plan remains obscure due to limited access to human embryos, while elucidating the mechanisms will advance knowledge of human development and contribute to regenerative medicine. The goal of my research is to reconstruct development of the human body axis and appendages using pluripotent stem cells (iPSC) and to illuminate general principles of pattern formation. I have used iPSC to successfully establish two 3D models recapitulating spatiotemporal features of human somite formation, which provide a valuable platform to decode mechanisms of body axis patterning with unprecedented resolution. Using these models, I found that active cell sorting underlies the antero-posterior (AP) polarity patterning of somites and put forward a novel framework explaining the formation of repeated structures along the body axis. In this proposal I will further utilize the in vitro models to expand and refine the cell sorting centered framework of somite AP patterning, as well as develop 3D models of human limb development to set up foundation for my future investigation of digit periodic patterning. In Aim1, I will investigate how cell sorting are coordinated with classical modules of somitogenesis. In the mentored phase, I will uncover roles of the segmentation clock in synchronizing the onset of cell sorting within a forming segment to ensure robust tissue-level patterning. In the independent phase, I will interrogate how somite size control is coordinated with AP pattern formation. Together these will bridge the novel finding of cell sorting with established concepts of somitogenesis. In Aim2, I will elucidate the molecular and cellular mechanisms of cell sorting and shine light on general cell biological principles of pattern formation. In parallel, I will set out to develop organoid models of human limb development using iPSCs. In Aim3, I will combine a preliminary 2D differentiation protocol of Limb bud cells in the Pourquie lab, the expertise in maintaining limb progenitors in vitro of the Tabin lab, and my specialty in 3D organoid culture to build 3D models recapitulating features of limb bud elongation and patterning. The proposed work will strengthen my knowledge of body axis development, equip me with expertise in limb development, and establish groundwork for my future career in elucidating periodic pattern formation during human development. Altogether, this proposal will provide valuable insights into the early development of human body plan and guide complex organ engineering to further dissect mechanisms and treat diseases.

在人体计划的开发过程中，如何实现精确的模式仍然不清楚， 有限地获得人类胚胎，同时阐明机制将促进人类胚胎发育的知识。 发展和促进再生医学。我的研究目标是重建 使用多能干细胞（iPSC）的人体轴和附件，并阐明 图案形成我已经使用iPSC成功地建立了两个3D模型， 体节形成的特征，为解读体轴机制提供了有价值的平台 以前所未有的分辨率形成图案。使用这些模型，我发现活跃的细胞分选是 体细胞的前后极性模式，并提出了一个新的解释框架 沿着身体轴线的重复结构。在这个建议中，我将进一步利用体外模型来扩大和 完善以细胞分选为中心的体节AP图案化框架，以及开发人体肢体的3D模型 发展，为今后数字周期图案的研究奠定基础。 在目标1中，我将研究细胞分选如何与体节发生的经典模块协调。在 在指导阶段，我将揭示分段时钟在同步细胞分选的开始中的作用。 以确保稳健的组织水平图案化。在独立阶段，我将询问如何体节 尺寸控制与AP图案形成相协调。这些将共同为细胞分选的新发现搭建桥梁 已经建立的体节发生的概念。在Aim 2中，我将阐明 细胞分选和照射模式形成的一般细胞生物学原理。与此同时，我将着手 使用iPSC开发人类肢体发育的类器官模型。在目标3中，我将结合联合收割机 Pourquie实验室的肢芽细胞分化方案，体外维持肢祖细胞的专业知识 Tabin实验室的，我在3D类器官培养方面的专长是建立重现肢芽特征的3D模型， 延伸和图案化。 建议的工作将加强我的身体轴发展的知识，装备我的专长， 肢体发育，并为我未来的职业生涯奠定基础，阐明周期性模式的形成， 人类发展总之，这一建议将为人类的早期发展提供有价值的见解 身体规划和指导复杂的器官工程，以进一步剖析机制和治疗疾病。