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.

项目概要 使用微流体技术的完全图案化的人类神经管模型 脊椎动物中枢神经系统（CNS）的发育始于神经管（NT）的形成 及其区域模式，沿头侧 (R)-尾侧 (C) 和背侧 (D)-生成神经元亚型 腹 (V) 轴。人类 NT 的区域模式是一个严格调控的过程，偏离该过程可能会 导致神经发育障碍，并可能在以后导致不同的神经和精神疾病 生活。由于人类接触的机会有限，人类 NT 的区域模式仍然不完全清楚。 胚胎组织。动物模型有助于了解人类中枢神经系统的发育 和相关疾病。然而，它们在揭示发展的一些基本方面方面有限， 人类独有的遗传学、病理学和疾病机制。基于干细胞的体外模型 人类神经系统发育，包括神经类器官和生物工程 NT 发育模型， 正在成为有前途的实验工具。然而，目前还没有基于干细胞的神经 开发模型能够沿着 3D 管状结构中的两个正交轴重现神经模式 几何学，NT 体内图案的标志。此外，现有的神经发育模型仅 概括了人类大脑或脊髓区域（但不是两者）发育的某些方面。 在我们的初步研究中，我们已经成功地利用了发展潜力和自我 人类多能干细胞 (hPSC) 的组织特性与微流体技术相结合，开发 第一个合成的、完全模式化的人类 NT 模型。使用该微流体平台，外源性 可以建立沿两个正交轴的形态发生素梯度以实现区域图案化 沿着 R-C 和 D-V 轴的微流体人类 NT 样结构，在脑样和脊髓样中 地区。这种微流体图案化的人类 NT 样结构表现出 NT 发育的许多特征， 包括管状几何形状，由神经元祖细胞包围的单个连续中央腔， 包括 HOX 基因在内的规范 R-C 和 D-V 区域标记的模式化表达，以及 神经中胚层祖细胞和峡组织者。因此，人体微流控技术的发展 NT 样结构密切模仿 NT 发育，首次提供类似体内的组织结构 具有一致的时空细胞分化和组织。 这项 R01 研究的目标是开发这种令人兴奋的微流体人类 NT 样模型（目标 1）和 利用其技术优势，研究不同外源形态发生素信号在神经中的作用 图案化（目标 2 和 3）。将进行遗传扰动和谱系追踪分析来研究人类 神经中胚层祖细胞发育（目标 2）。在目标 3 中，我们进一步致力于实现 D-V 模式的人类 具有前脑或脊髓特性的类似 NT 的结构，并使用这些结构来概括 皮质发育过程中区域间细胞相互作用和兴奋与抑制相互作用。