Deciphering Principles of Human Embryonic Patterning in Development and Disease

项目摘要/摘要 发育生物学中的基本问题围绕着理解分子和 调节血统规范和分化的细胞事件。没有比这更令人费解的了 与早期人类胚胎相比，由于过去的技术和伦理原因，早期人类胚胎在实验上仍然难以解决 原肠形成的开始-人体的三个主要生殖层被指定为 胚胎上皮细胞组织。我和其他人之前开发了基于干细胞的系统 概述了三维(3D)哺乳动物体外发育的关键方面，为更多的 完全了解管理胚胎发育的具体步骤。在这里，我提出了一种创新的 新的实验方法，使用人类多能干细胞(HPSCs)，结合尖端技术 用于可控、高效和可扩展的人类外胚层发育模型的生物工程技术 在试管中。该模型将使研究人类生殖层分化轨迹和时间动力学成为可能 以正确的3D构象。我将结合遗传学、成像和单细胞多细胞- 组学与高通量计算分析相结合，以定义关键的分子和细胞事件 在不同的条件下调节发育模式。我将进一步调查这些事件是如何 由细胞新陈代谢和营养可获得性控制，这对理解 许多人类疾病的早期胚胎起源。我的提议将开启一个全新而强大的 剖析人类发育遗传学基本的、相互关联的原则的实验范式 在早期胚胎阶段，否则是无法接触到的。这一新知识还将直接为以下努力提供信息 高效地从干细胞产生成熟的组织和细胞类型，用于疾病建模和细胞治疗。 最终，这些发现将对可能预防不良妊娠结局至关重要，提供了一种 了解发育障碍背后的细胞和分子机制的独特机会 先天病症。

Project Summary/Abstract Fundamental questions in developmental biology revolve around understanding the sequence of molecular and cellular events that regulates lineage specification and differentiation. Nowhere is this more poorly understood than in the early human embryo, which remains experimentally intractable for technical and ethical reasons past the onset of gastrulation - the point when the three major germ layers of the human body are specified within the embryonic epiblast tissue. Myself and others have previously developed stem cell-based systems that recapitulate key aspects of 3-dimensional (3D) mammalian development in vitro, paving the way to a more complete understanding of the specific steps that govern embryonic development. Here, I propose an innovative new experimental approach, using human pluripotent stem cells (hPSCs), combined with cutting-edge bioengineering technologies for a controllable, efficient and scalable modeling of human epiblast development in vitro. This model will permit studying the human germ layer differentiation trajectory and temporal dynamics in the correct 3D-conformation. I will combine state-of-the-art tools in genetics, imaging, and single-cell multi- omics in tandem with high-throughput computational analyses to define the key molecular and cellular events that regulate developmental patterning under different conditions. I will further investigate how these events are controlled by cellular metabolism and nutrient availability, which has important implications for understanding the early embryonic origin of numerous human diseases. My proposal will open up a completely new and powerful experimental paradigm to dissect the fundamental, inter-connected, principles of human developmental genetics at early embryonic stages that are otherwise inaccessible. This new knowledge will also directly inform efforts to efficiently generate mature tissues and cell types from stem cells for disease modeling and cell therapies. Ultimately, these findings will be critical for possible prevention of adverse pregnancy outcomes, offering a unique opportunity to understand the cellular and molecular mechanisms behind developmental disorders and congenital pathologies.