Modeling NDE1 function in dysregulated brain development using a microfluidic CNS model

使用微流体中枢神经系统模型模拟 NDE1 在大脑发育失调中的功能

• 批准号: 10666902
• 负责人: Jianping Fu
• 金额: $ 20.64万
• 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: 2023-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666902
• 关键词: 3-Dimensional; Address; Animal Model; Benchmarking; Biological Models; Brain; Brain Diseases; Brain region; Cell division; Cells; Central Nervous System; DNA Sequence Alteration; Data; Development; Developmental Process; Disease; Disease model; Dorsal; Elements; Embryo; Etiology; Exhibits; Genes; Genetic; Genetic study; Geometry; Goals; Growth; Health; Human; Individual; Knock-out; Knockout Mice; Measurement; Mediating; Microcephaly; Microfluidic Microchips; Microfluidics; Midbrain structure; Modeling; Mus; Mutation; Neural Tube Development; Neural tube; Nuclear; Organoids; Pathology; Pattern; Positioning Attribute; Pregnancy; Prevalence; Process; Property; Prosencephalon; Proteins; Protocols documentation; Reproducibility; Research; Role; Societies; Spinal Cord; Structure; Technology; Tissues; Tubular formation; brain abnormalities; cell motility; comparative; disease phenotype; genome sequencing; hindbrain; human model; human pluripotent stem cell; in vitro Model; in vivo; innovation; model development; morphogens; mouse model; mutant; mutant mouse model; nerve stem cell; neural network; neural patterning; neurodevelopment; novel; personalized medicine; public health relevance; self organization; single-cell RNA sequencing; spatiotemporal; stem cells; tool; transcriptome

Project Summary Modeling NDE1 function in dysregulated brain development using a microfluidic CNS model Development of the vertebrate central nervous system (CNS) begins with the formation of neural tube (NT) and its regional patterning to generate the forebrain, midbrain, hindbrain, and spinal cord. Regional patterning of the brain is a tightly regulated developmental process, deviation from which can result in neurodevelopmental brain diseases. Multiple causes are associated with neurodevelopmental brain diseases, including genetic, environmental, infectious, and traumatic factors. Even though the precise etiology of neurodevelopmental brain diseases remains largely unknown, the genetic components of neurodevelopmental brain diseases have been increasingly deciphered with the advent of personalized medicine. However, detailed pathophysiological mechanisms of neurodevelopmental brain diseases remain challenging to study, due to limited access to human CNS tissues. Animal models have been instrumental in understanding human neurodevelopment and associated disorders. However, they are limited in revealing some of the most fundamental aspects of development, genetics, pathology, and disease mechanisms that are unique to humans. Stem cell-based in vitro models of human neurodevelopment are emerging as promising experimental tools. However, the controllability and reproducibility of these models remain suboptimal. Furthermore, none of the current neurodevelopment models is capable of recapitulating regional patterning of the brain faithfully in a 3D tubular geometry, a hallmark of neurodevelopment. The goal of this R21 project is to specifically address this significant technological need, by using human pluripotent stem cells (hPSCs) to develop a human brain development model that can faithfully recapitulate regional brain patterning. Importantly, we propose to apply this model to study the function of nuclear distribution element 1 (NDE1), a gene implicated in a wide range of neurodevelopmental conditions, including microcephaly (a small brain), microlissencephaly (a small brain with a simplified gyral pattern), or microhydranencephaly (a more severe presentation). In our preliminary study, we have successfully leveraged the developmental potential and self-organizing property of hPSCs in conjunction with innovative microfluidics to develop the first of its kind, synthetic, fully patterned human NT model. Our preliminary data from brain organoids generated from NDE1-knockout (KO) hPSCs further show that NDE1-KO brain organoids exhibit reduced growth and gyrification and furthermore show abnormal brain regionalization. Thus, our preliminary data suggest a novel and previously unexplored mechanism involving dysregulated brain regionalization in NDE1-mediated microcephaly. In this proposal, we propose to first extend the microfluidic patterned human NT model to recapitulate brain regionalization (Aim 1). We will then utilize this controllable human brain development model and a Nde1-KO mouse model to study the role of NDE1 mutations in dysregulated brain regionalization (Aim 2).

项目摘要 用微流控中枢神经系统模型模拟脑发育异常中NDE1的功能 脊椎动物中枢神经系统的发育始于神经管的形成和 它的区域模式产生前脑、中脑、后脑和脊髓。区域格局的形成 大脑是一个受到严格控制的发育过程，偏离这个过程会导致神经发育。 脑部疾病。多种原因与神经发育性脑部疾病有关，包括遗传， 环境因素、传染病因素和创伤因素。即使神经发育的确切病因学 脑部疾病在很大程度上仍不清楚，神经发育性脑部疾病的遗传成分 随着个性化医学的出现而日益被破译。然而，详细的病理生理 神经发育性脑疾病的机制研究仍然具有挑战性，因为获得的机会有限 人中枢神经系统组织。动物模型在理解人类神经发育和 伴发的疾病。然而，它们在揭示一些最基本的方面方面受到限制。 人类独有的发育、遗传学、病理学和疾病机制。干细胞为基础的 人类神经发育的体外模型正在成为很有前途的实验工具。然而， 这些模型的可控性和重复性仍然不是最优的。此外，没有一个当前的 神经发育模型能够在3D管状结构中忠实地再现大脑的区域模式 几何学，神经发育的标志。 该R21项目的目标是通过使用以下方法来具体解决这一重要的技术需求 人类多能干细胞(HPSCs)开发出一种人脑发育模型，可以忠实地 概括一下大脑的局部模式。重要的是，我们建议应用这一模型来研究 核分布元件1(NDE1)是一种与一系列神经发育疾病有关的基因， 包括小头症(小脑)、小脑(具有简化的脑回模式的小脑)，或 小脑积水(一种更严重的表现)。在我们的初步研究中，我们成功地利用 HPSCs与创新微流控相结合的发展潜力和自组织特性 开发出首个人工合成的全图案人体NT模型。我们来自大脑的初步数据 从NDE1基因敲除(KO)hPSCs产生的有机类化合物进一步表明NDE1-KO脑有机类化合物表现出 生长和脑回变减慢，并进一步显示大脑区域化异常。因此，我们初步的 数据表明，一种新的、以前未被探索的机制涉及到脑功能失调 NDE1介导的小头畸形。在这个方案中，我们建议首先扩展微流控图案的人体 NT模型重现脑区划(目标1)。然后我们将利用这个可控的人脑 研究NDE1基因突变在脑功能失调中作用的发育模型和NDE1-KO小鼠模型 区域化(目标2)。