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 使用微流体CNS模型在失调的大脑发育中建模NDE1功能 脊椎动物中枢神经系统（CNS）的发展始于形成神经管（NT）和 它的区域模式以产生前脑，中脑，后脑和脊髓。区域构图 大脑是一个严格调节的发育过程，偏差会导致神经发育 脑部疾病。多种原因与神经发育脑疾病有关，包括遗传， 环境，传染性和创伤因素。即使神经发育的精确病因 脑部疾病在很大程度上尚不清楚，神经发育脑疾病的遗传成分具有 随着个性化医学的出现，越来越多地破译。但是，详细的病理生理学 神经发育脑部疾病的机制仍然具有挑战性 人类中枢神经系统组织。动物模型对了解人类神经发育和 相关疾病。但是，它们在揭示一些最根本的方面受到限制 人类独有的发展，遗传学，病理和疾病机制。基于干细胞IN 人类神经发育的体外模型正在成为有前途的实验工具。但是， 这些模型的可控性和可重复性仍然是最佳的。此外，当前都没有 神经发育模型能够在3D管状中忠实地概括大脑的区域模式 几何，神经发育的标志。 该R21项目的目标是通过使用 人类多能干细胞（HPSC）开发可以忠实的人脑发育模型 概括区域大脑模式。重要的是，我们建议应用此模型来研究 核分布元件1（NDE1），该基因与广泛的神经发育条件有关， 包括小头畸形（一个小大脑），微卵形畸形（具有简化陀螺模式的小大脑）或 微水脑（更严重的表现）。在我们的初步研究中，我们成功利用了 HPSC的发展潜力和自组织特性与创新的微流体相结合 为了开发同类的第一个，合成的，完全构图的人类NT模型。我们的大脑初步数据 NDE1-KNOCKOUT（KO）HPSC产生的类器官进一步表明NDE1-KO脑器官展示 降低的生长和回旋以及进一步显示出异常的大脑区域化。因此，我们的初步 数据表明，一种新颖且以前未开发的机制，涉及大脑失调的大脑区域化 NDE1介导的小头畸形。在此提案中，我们建议首先扩展微流体图案化的人类 概括大脑区域化的NT模型（AIM 1）。然后，我们将利用这种可控的人脑 开发模型和NDE1-KO小鼠模型研究NDE1突变在大脑失调失调中的作用 区域化（目标2）。