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.

项目摘要阿尔茨海默氏病（AD）是一种进行性神经退行性脑疾病，会损害记忆和认知功能。它是65岁以上老年人中最常见的痴呆症，据估计， 580万美国人可能患有AD引起的痴呆症。尽管AD的发病机理尚不清楚，但异常淀粉样蛋白β（Aβ）斑块的沉积物和整个大脑的高磷酸化tau蛋白被认为是在神经炎症，突触丧失和神经元细胞死亡中发挥作用。而Aβ似乎扩散假设磷酸化的tau蛋白是在突触连接的神经元之间传播的。最近的研究表明，Aβ的存在可能会增加传播的速率，这可能是由于神经炎症作用。此外，神经炎症可能直接诱导或加剧Aβ和TAU 蛋白质病，从而更糟的神经炎症并导致突触和神经元的进一步丧失，创建一个可能促进疾病进展的恶性循环。但是，基于的确切机制传播磷酸化的tau及其与神经炎症的相互作用仍然难以捉摸。区分这些在体内彼此之间存在许多混杂信号的彼此的复杂因素极具挑战性。因此，需要新方法来弥合这一差距，以揭示基本机制异常蛋白质和神经炎症的转运加速了AD的发展。为了满足这一需求，我们将使用一种新型三文化的体外平台使用微流体的体外平台（原发性神经元，星形胶质细胞，小胶质细胞）大鼠的神经炎症模型，这些模型已作为一部分开发 PIS的当前R03奖。微流体平台由两个物理上不同的培养室组成（例如，初级和次要），对应于微通道互连的近端和远端解剖区域这允许两种培养物之间的合成连通性。这种行政补充的直接目标是为了研究神经胶质细胞和感染对异常Tau蛋白的运输的贡献。具体来说，我们将（i）确定Aβ和磷酸化的人tau（由转染神经元表达）对三培养模型中的神经炎症，（ii）采用微流体平台将其脱离 Aβ添加本身以及Aβ触发的神经炎症沿着轴突的tau传播连接两种培养物保持在初级和次级室内。预计此处描述的试点研究（i）确定致病会议Aβ添加到培养和/或人tau表达中的影响通过转染神经元对神经炎症的转染，（ii）将致病性Aβ本身或其其影响的影响解散相应的神经炎性细胞因子特征在致病性TAU传播的轴突谱和（iii）的传播中为神经炎症与轴突之间的相互作用的未来机械研究建立基础在阿尔茨海默氏病的背景下，致病性tau的运输。