Interplay of Neuroinflammation and Tau Transport in a Microfluidic Primary Neural Cell Tri-Culture Model

微流体原代神经细胞三培养模型中神经炎症和 Tau 转运的相互作用

基本信息

批准号：
10289580
负责人：
Erkin Seker
金额：
$ 7.14万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10289580
关键词：
Achievement Address Administrative Supplement Alzheimer&apos s Disease American Amyloid beta-Protein Anatomy Anti-Inflammatory Agents Apoptosis Astrocytes Award Axon Axonal Transport Beta Cell Biochemical Biological Assay Brain Brain Diseases Brain region Cessation of life Chemicals Collaborations Complex Dementia Deposition Diffuse Disease Progression Distal Elderly Engineering Ensure Fluorescence Microscopy Formulation Foundations Future Goals Health Histologic Human Hydrostatic Pressure In Vitro Inflammation Interneurons Lead Liquid substance Memory impairment Methodology Microfabrication Microfluidics Microglia Modeling Molecular Conformation Monitor Morphology Nerve Degeneration Neuroglia Neurons Pathogenesis Pathogenicity Pathologic Pilot Projects Play Protein Conformation Proteins Proteomics Rattus Resistance Role Route Senile Plaques Signal Transduction Synapses Tissues Transfection axon growth cognitive function cytokine human old age (65+)hyperphosphorylated tau in vivo insight miniaturize neuroinflammation neuron loss neutralizing antibody novel rapid testing relating to nervous system response tau Proteins tau expression tau mutation tau-1 therapeutic evaluation uptake

项目摘要

PROJECT SUMMARY Alzheimer’s Disease (AD) is a progressive neurodegenerative brain disorder that impairs memory and cognitive functions. It is the most common dementia among older adults over age 65 and it is estimated that more than 5.8 million Americans may have dementia caused by AD. While the pathogenesis of AD is unclear, abnormal deposits of amyloid-β (Aβ) plaques and hyperphosphorylated tau proteins throughout the brain are thought to play a role in neuroinflammation, synapse loss, and neuronal cell death. While Aβ appears to spread in a diffuse manner, phosphorylated tau proteins are hypothesized to propagate between synaptically connected neurons. Recent studies suggest that the presence of Aβ may increase the rate of propagation, potentially due to neuroinflammatory effects. In addition, neuroinflammation may directly induce or exacerbate Aβ and tau proteinopathies, thereby worsen neuroinflammation and leading to further loss of synapses and neurons, creating a vicious cycle that likely promotes disease progression. However, the exact mechanisms that underlie propagation phosphorylated tau and its interplay with neuroinflammation remain elusive. Distinguishing these complex factors from each other in vivo, where numerous confounding signals exist, is extremely challenging. There is, therefore, a need for new methodologies to bridge this gap for revealing the underlying mechanisms by which transport of aberrant proteins and neuroinflammation accelerate the progression of AD. In order to address this need, we will employ a microfluidic in vitro platform in combination with a novel tri-culture (primary neuron, astrocyte, microglia) rat model of neuroinflammation that have been developed as part of the PIs’ current R03 award. The microfluidic platform consists of two physically distinct culture chambers (e.g., primary and secondary), corresponding to proximal and distal anatomic regions interconnected by microchannels that allow synaptic connectivity between the two cultures. The immediate goal of this administrative supplement is to study the contribution of glial cells and inflammation to the transport of abnormal tau proteins. Specifically, we will (i) determine the influence of Aβ and phosphorylated human tau (expressed by transfected neurons) on neuroinflammation in the tri-culture model, and (ii) employ the microfluidic platform to decouple the influences of Aβ addition itself and the Aβ-triggered neuroinflammation on tau propagation along the axonal tracts connecting the two cultures maintained in the primary and secondary chambers. The pilot study described here is expected to (i) identify the influence of pathogenic conformations Aβ added to the culture and/or human tau expression by transfected neurons on neuroinflammation, (ii) decouple the influence of pathogenic Aβ itself or its corresponding neuroinflammatory cytokine profile on propagation of pathogenic tau via axonal tracts, and (iii) establish the foundation for future mechanistic studies of the interplay between neuroinflammation and axonal transport of pathogenic tau in the context of Alzheimer’s Disease.

项目概要阿尔茨海默病 (AD) 是一种进行性神经退行性脑部疾病，会损害记忆和认知它是 65 岁以上老年人中最常见的痴呆症，据估计超过 580万美国人可能患有AD引起的痴呆，而AD的发病机制尚不清楚，不正常。 β 淀粉样蛋白 (Aβ) 斑块和过度磷酸化的 tau 蛋白在整个大脑中的沉积被认为 Aβ 在神经炎症、突触损失和神经元细胞死亡中发挥作用，而 Aβ 似乎呈弥漫性扩散。以这种方式，磷酸化的 tau 蛋白被捕获并在突触连接的神经元之间传播。最近的研究表明，Aβ 的存在可能会增加传播速度，这可能是由于此外，神经炎症可能直接诱导或加剧 Aβ 和 tau 蛋白的产生。蛋白质病，从而加剧神经炎症并导致突触和神经元进一步丧失，造成一个可能促进疾病进展的恶性循环，然而，其背后的确切机制。磷酸化 tau 蛋白的传播及其与神经炎症的相互作用仍然难以区分。在体内存在大量混杂信号的情况下，研究彼此之间的复杂因素极具挑战性。因此，需要新的方法来弥补这一差距，以揭示潜在的机制异常蛋白质的转运和神经炎症加速了 AD 的进展。为了满足这一需求，我们将采用微流体体外平台与新型三培养相结合（原代神经元、星形胶质细胞、小胶质细胞）神经炎症大鼠模型，已作为 PI 当前的 R03 奖微流体平台由两个物理上不同的培养室组成（例如，主要和次要），对应于通过微通道互连的近端和远端解剖区域允许两种文化之间的突触连接。本行政补充的近期目标。具体来说，是研究神经胶质细胞和炎症对异常 tau 蛋白运输的贡献。我们将 (i) 确定 Aβ 和磷酸化人 tau（由转染的神经元表达）对三培养模型中的神经炎症，以及（ii）利用微流体平台来解耦以下因素的影响 Aβ 添加本身以及 Aβ 触发的 tau 蛋白沿着连接的轴突束传播的神经炎症预计此处描述的试点研究将在初级室和次级室中维持两种培养物。 (i) 确定添加到培养物中的致病构象 Aβ 和/或人类 tau 表达的影响通过转染神经元对神经炎症的影响，(ii) 解耦致病性 Aβ 本身或其影响致病性 tau 蛋白通过轴突束传播的相应神经炎症细胞因子谱，以及 (iii) 为未来神经炎症和轴突之间相互作用的机制研究奠定基础阿尔茨海默病中致病性 tau 蛋白的转运。