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

基本信息

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
项目类别：
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

项目摘要

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).

项目概要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脊椎动物中枢神经系统（CNS）的发育始于神经管（NT）的形成和它的区域模式产生前脑、中脑、后脑和脊髓。区域格局大脑是一个受到严格调控的发育过程，偏离该过程可能会导致神经发育障碍脑部疾病。多种原因与神经发育性脑疾病有关，包括遗传、环境、传染性和创伤性因素。尽管神经发育的确切病因学脑部疾病仍然很大程度上未知，神经发育性脑部疾病的遗传成分已经随着个性化医疗的出现，这一问题被越来越多地解读。然而，详细的病理生理学由于获取途径有限，神经发育性脑疾病的机制仍然难以研究人类中枢神经系统组织。动物模型有助于理解人类神经发育和相关疾病。然而，它们在揭示一些最基本的方面方面是有限的。人类特有的发育、遗传学、病理学和疾病机制。以干细胞为基础人类神经发育的体外模型正在成为有前途的实验工具。然而，这些模型的可控性和再现性仍然不够理想。此外，目前没有任何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。 R21 项目的目标是通过使用人类多能干细胞（hPSC）开发人类大脑发育模型，可以忠实地概括区域大脑模式。重要的是，我们建议应用该模型来研究核分布元件 1 (NDE1)，一种与多种神经发育状况有关的基因，包括小头畸形（小大脑）、小无脑畸形（具有简化脑回模式的小大脑），或微积水脑畸形（更严重的表现）。在我们的初步研究中，我们成功地利用了 hPSC 的发展潜力和自组织特性与创新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。我们来自大脑的初步数据由 NDE1 敲除 (KO) hPSC 生成的类器官进一步表明，NDE1-KO 脑类器官表现出生长和回转减少，此外还显示出异常的大脑区域化。于是，我们初步数据表明，存在一种新颖且以前未探索过的机制，涉及大脑区域化失调 NDE1 介导的小头畸形。在这个提案中，我们建议首先扩展微流体图案的人体 NT 模型概括大脑区域化（目标 1）。然后我们就利用这个可控的人脑发育模型和 Nde1-KO 小鼠模型，用于研究 NDE1 突变在大脑失调中的作用区域化（目标 2）。