Developing organoid model to study active folding in a human genetic context

开发类器官模型来研究人类遗传背景下的主动折叠

基本信息

批准号：
9810045
负责人：
Sebastian J Streichan
金额：
$ 19.38万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9810045
关键词：
3-Dimensional Actomyosin Anatomy Anencephaly Animal Model Architecture Atlases Behavior Biological Models Brain Cell Culture Techniques Cells Cellular Structures Characteristics Complex Controlled Environment Cytoskeleton Data Analyses Defect Development Developmental Biology Dimensions Disease Embryonic Development Epithelium Extracellular Space Failure Foundations Gene Expression Profile Genetic Human Human Development Human Genetics Image In Vitro Investigation Kinetics Light Live Birth Lung Mechanics Methods Microfluidics Modeling Molecular Natural regeneration Neural Tube Closure Neural tube Neurons New Territories Nonmuscle Myosin Type IIA Organ Organism Organogenesis Organoids Pattern Pharmaceutical Preparations Physics Positioning Attribute Process Protocols documentation Reproducibility Role SHH gene Severities Shapes Signal Transduction Spinal Dysraphism Stem cells Structure Technology Tissues Tongue Tube Vertebral column base behavior measurement cell behavior cell type convergent extension design flexibility human model imaging approach in vivo insight matrigel morphogens neural model neuroepithelium novel overexpression prevent programs quantitative imaging relating to nervous system small molecule smoothened signaling pathway stillbirth transcription factor

项目摘要

ABSTRACT Organ shape is critical for the organism to function properly. For the example neural tube, early formation defects may have devastating consequences to the body. A fundamental building principal in embryogenesis is first organizing cells into simple structures, such as a closed sheet surrounding a lumen. Cells then undergo well-concerted programs to reconfigure tissue shape, or fold out of the plane of the sheet. In plane flows elongate one axis while shortening another during convergent extension. Folding in contrast dramatically changes tissue form, to endow the organ with its characteristic shape. In neural tube closure, an initially flat sheet of cells undergoes two sequential bending steps to fold up into a cylindrical tube surrounding a single lumen. A large body of work uncovered fundamental insights on fate determination. Yet, mechanisms controlling shape, particularly folding in the context of early human development, remain poorly understood. At the cellular level genetic patterning coordinates behaviors over long distances to instruct active forces that underlie folding. Before distilling a physical picture of how forces fold tissue, we need to identify behaviors leading to folding, and characterize coordination. Organoids harbor promise for studying early development, disease and regeneration in tightly controlled environments and human genetic background. However, progress is hampered by technical limitations, due to irregular patterning, and shape. This proposal seeks to develop a new strategy for designing robust models to study basic mechanisms of human organogenesis. Using micro patterning we successfully generated highly reproducible organoids with set size, shape, and formation kinetics. Our approach features flat rectangular shaped sheets that spontaneously fold out of the plane, and seamlessly close along the short dimension, generating a tube. The resulting platform is high throughput, compatible with live imaging, and well suited for quantitative data analysis strategies. This setup recapitulates major steps during neural tube closure in animal model systems, such as hinge formation and a zippering mechanism with actomyosin cable accompanying closure. Here we aim to adapt existing protocols to our platform to 1) induce formation of neuroepithelium and more closely model neural tube formation. We then aim to 2) assemble a quantitative atlas of cell behaviors observed during closure. Finally, we aim to 3) enter new territory and generate anatomical axes spanning the organoid tube using microfluidics. Our approach opens up a new path towards controlled models of human organogenesis, using the neural tube as example.

抽象的器官形状对于生物体正常运行至关重要。对于示例神经管，早期形成缺陷可能对身体产生毁灭性后果。胚胎发生的基本建筑物主要是首先将细胞整理成简单的结构，例如围绕管腔的封闭板。然后经过细胞良好的节目以重新配置组织形状，或从纸张平面上折叠。在平面流中在收敛延伸过程中缩短另一个轴，同时缩短另一个轴。对比度折叠很大改变组织形式，以赋予器官特征形状。在神经管闭合中，最初是平坦的一组细胞经历两个顺序的弯曲步骤，将围绕一个单一的圆柱管折叠成一个流明。大量的工作发现了对命运决定的基本见解。但是，机制控制形状，尤其是在早期人类发展的背景下折叠的形状，对形状仍然很少理解。在细胞水平上折叠基础。在蒸馏出强力如何折叠组织的物理图片之前，我们需要识别行为导致折叠并表征协调。类器官港口有望研究早期发展，在紧密控制的环境和人类遗传背景下的疾病和再生。然而，由于模式不规则和形状，技术局限性受到了技术限制的阻碍。该提议试图制定一种新的策略来设计强大的模型来研究人体器官发生的基本机制。使用微模式，我们成功地生成了具有设定尺寸，形状和形状和编队动力学。我们的方法具有扁平矩形形状的床单平面，并沿短尺寸无缝闭合，生成管。最终的平台很高吞吐量，与实时成像兼容，非常适合定量数据分析策略。此设置在动物模型系统中，概括神经管闭合期间的主要步骤，例如铰链形成和A 带有肌动蛋白电缆的拉链机制伴随闭合。在这里，我们旨在调整现有协议我们的平台1）诱导神经上皮的形成，并更紧密地模拟神经管的形成。然后我们目的是2）组装闭合过程中观察到的细胞行为的定量地图集。最后，我们的目标是3）输入新区域并生成使用微流体跨器官管的解剖轴。我们的方法以神经管为例，为受控人体器官发生模型打开了新的路径。