A Fully Patterned Human Neural Tube Model Using Microfluidics

使用微流体技术的完全图案化的人类神经管模型

• 批准号: 10732812
• 负责人: Jianping Fu
• 金额: $ 52.34万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10732812
• 关键词: 3-Dimensional; Address; Anatomy; Animal Model; Architecture; Biological Assay; Biomedical Engineering; Brain; Cell Differentiation process; Cell Therapy; Central Nervous System; Central Nervous System Diseases; Data; Development; Diagnosis; Disease; Disease model; Dorsal; Ectoderm; Embryo; Embryology; Event; Exhibits; Foundations; Genetic; Geometry; Goals; Homeobox Genes; Human; Knowledge; Life; Measurement; Mental disorders; Microfluidics; Midbrain structure; Modeling; Molecular; Nerve Tissue; Neural Tube Development; Neural tube; Neurodevelopmental Disorder; Neuronal Differentiation; Neurons; Organoids; Pathology; Pattern; Pharmaceutical Preparations; Play; Prevention; Process; Property; Prosencephalon; Research; Role; Signal Transduction; Specific qualifier value; Spinal Cord; Stem Cell Development; Structure; Techniques; Testing; Time; Tissues; Toxic effect; Tubular formation; differentiation protocol; embryo tissue; high throughput screening; hindbrain; human model; human pluripotent stem cell; human stem cells; in vitro Model; in vivo; innovation; morphogens; nerve stem cell; nervous system development; nervous system disorder; neural; neural patterning; neural plate; neurodevelopment; neuroregulation; notochord; progenitor; public health relevance; rational design; self organization; spatiotemporal; stem cell model; stem cell therapy; stem cells; success; tool

Project Summary A Fully Patterned Human Neural Tube Model Using Microfluidics The development of vertebrate central nervous system (CNS) begins with the formation of neural tube (NT) and its regional patterning to generate neuronal subtypes along the rostral (R)-caudal (C) and dorsal (D)- ventral (V) axes. Regional patterning of the human NT is a tightly regulated process, deviation from which can result in neurodevelopmental disorders and may lead to distinct neurological and psychiatric diseases later in life. Regional patterning of the human NT remains incompletely understood due to limited access to human embryonic tissues. Animal models have been instrumental in understanding the development of human CNS and associated disorders. However, they are limited in revealing some fundamental aspects of development, genetics, pathology, and disease mechanisms that are unique to humans. Stem cell-based in vitro models of human nervous system development, including neural organoids and bioengineered NT development models, are emerging as promising experimental tools. However, none of the current stem cell-based neural development models is capable of recapitulating neural patterning along two orthogonal axes in a 3D tubular geometry, the hallmark of NT patterning in vivo. Furthermore, the existing neural development models only recapitulate certain aspects of the development of either human brain or spinal cord regions but not both. In our preliminary study, we have successfully leveraged the developmental potential and self- organizing property of human pluripotent stem cells (hPSCs) in conjunction with microfluidics to develop the first of its kind, synthetic, fully patterned human NT model. Using this microfluidic platform, exogenous morphogen gradients along two orthogonal axes can be established to achieve regional patterning of the microfluidic human NT-like structure along both the R-C and D-V axes, in both brain-like and spinal cord-like regions. This microfluidic patterned human NT-like structure exhibits many hallmarks of NT development, including a tubular geometry, a single continuous central lumen enclosing by neuronal progenitor cells, patterned expression of canonical R-C and D-V regional markers including HOX genes, and the emergence of neural mesodermal progenitors and the isthmic organizer. Thus, the development of the microfluidic human NT-like structure closely mimics NT development, offering for the first time an in vivo-like tissue architecture with consistent spatiotemporal cell differentiation and organization. The goal of this R01 research is to develop this exciting microfluidic human NT-like model (Aim 1) and leverage its technical advantages to study the roles of different exogenous morphogen signals in neural patterning (Aim 2 & 3). Genetic perturbations and lineage tracing assays will be conducted to study human neural mesodermal progenitor development (Aim 2). In Aim 3 we further aim to achieve D-V patterned human NT-like structures with either forebrain or spinal cord identities and use these structures to recapitulate interregional cellular interactions and excitation-to-inhibition interplays during cortical development.

项目摘要 使用微流体的完全模式的人类神经管模型 脊椎动物中枢神经系统（CNS）的发展始于形成神经管（NT） 及其区域模式以产生沿rostral（r）-caudal（c）和背（d）的神经元亚型 - 腹侧（V）轴。人类NT的区域模式是一个严格调节的过程，偏差可以从中 导致神经发育障碍，后来可能导致明显的神经和精神病 生活。人类NT的区域模式由于有限的人类机会而尚未完全理解 胚胎组织。动物模型有助于理解人类中枢神经系统的发展 和相关的疾病。但是，它们在揭示发展的某些基本方面受到限制， 人类独有的遗传学，病理和疾病机制。基于干细胞的体外模型 人类神经系统的发展，包括神经器官和生物工程的NT开发模型， 正在成为有希望的实验工具。但是，当前基于干细胞的神经没有 开发模型能够沿着3D管状的两个正交轴概括神经模式 几何，在体内图案的标志。此外，现有的神经发展模型 概括人脑或脊髓区域发展的某些方面，但并非两者兼而有之。 在我们的初步研究中，我们成功利用了发展潜力和自我 组织多能干细胞（HPSC）与微流体的组织特性 首先，合成的，完全构图的人类NT模型。使用此微流体平台，外源 可以建立沿两个正交轴的形态学梯度，以实现区域模式 沿R-C和D-V轴的微流体人类NT样结构，脑样和脊髓状的结构 地区。这种微流体图案化的人类NT样结构具有NT发展的许多标志， 包括管状几何形状，一个由神经元祖细胞封闭的单个连续中央管腔， 包括HOX基因在内的规范R-C和D-V区域标记的图案表达，以及出现 神经中胚层祖细胞和峡部组织者。因此，微流体人类的发展 NT状结构紧密模仿NT的开发，首次提供体内的组织结构 具有一致的时空细胞分化和组织。 这项R01研究的目的是开发这种令人兴奋的微流体人类NT样模型（AIM 1）和 利用其技术优势研究不同的外源形态学信号在神经中的作用 图案（AIM 2和3）。遗传扰动和谱系追踪测定将进行研究 神经中胚层祖细胞发展（AIM 2）。在目标3中，我们进一步旨在实现D-V图案的人 具有前脑或脊髓身份的NT样结构，并使用这些结构概括 皮质发育过程中区域间细胞相互作用和激发对抑制作用的互动。