High-Throughput In Vitro Analyses of Trauma-Induced Tauopathy

创伤引起的 Tau 病的高通量体外分析

• 批准号: 10647481
• 负责人: PATRICK W ALFORD
• 金额: $ 21.51万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647481
• 关键词: Affect; Axon; Cell Culture Techniques; Cells; Craniocerebral Trauma; Data; Dendritic Spines; Devices; Drug Screening; Functional disorder; Goals; Image; In Vitro; Infrastructure; Injury; Link; Measures; Mechanics; Methods; Microtubules; Modeling; Mus; Nervous System Trauma; Neurodegenerative Disorders; Neuronal Injury; Neurons; Outcome; Pathology; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacological Treatment; Stretching; Swelling; Synapses; System; TBI Patients; Tauopathies; Testing; Trauma; Work; chronic traumatic encephalopathy; design; drug discovery; efficacy testing; experimental study; fluorescence imaging; high throughput screening; in vitro Model; inhibitor; insight; machine learning algorithm; mild traumatic brain injury; patch clamp; pharmacologic; scale up; screening; tau Proteins

PROJECT SUMMARY Chronic traumatic encephalopathy (CTE) is a neurodegenerative disease associated with repeated mild traumatic brain injury (TBI). CTE is among the many neurodegenerative diseases characterized as tauopathies, wherein the protein tau, which is usually associated with microtubules in the axons of neurons, becomes separated from microtubules, initiating a degenerative cascade and leading to eventual neurofunctional loss. There are currently no pharmacological treatments available for CTE patients, so any treatment that could limit or reverse tau-associated dysfunction would have an important impact on TBI patient outcomes. Moreover, given the similarity between CTE and other tauopathies, insights into CTE treatment could be broadly applicable to other common neurodegenerative diseases. We have recently developed an in vitro model that directly links mechanical injury to tau pathology in cultured neurons. One notable outcome from our prior studies is that we found that synaptic dysfunction (measured using patch clamp) is correlated with mislocalization of tau to dendritic spines (measured using fluorescent imaging). This result suggests that this relatively simple image-based readout could be used for screening the effects of pharmacological agents on the functional progression of trauma-induced tauopathy. There are currently no high-throughput in vitro models for screening the effects of pharmaceuticals on trauma-induced tauopathy. We have developed several cell stretching systems, both for neurotrauma and other applications. However, all of these systems are far too low-throughput for performing drug discovery studies. Thus, our goal is to scale up our current in vitro neurotrauma model to a high-enough throughput for drug screening studies. We will design a new stretchable multi-well plate for neuronal cell culture and a new high strain rate stretcher that can apply trauma-like loads to the cells in the plate. In addition, we will employ machine learning based algorithms to quickly and efficiently analyze the data collected from our new device. Finally, we will use the device to test inhibitors known to be effective against other aspects of neuronal injury affect tau mislocalization

项目摘要 慢性创伤性脑病（CTE）是一种神经退行性疾病， 反复轻度创伤性脑损伤（TBI）。CTE是许多神经退行性疾病之一， 其特征在于tau蛋白病，其中通常与 神经元轴突中的微管与微管分离， 退化级联并导致最终的神经功能丧失。目前没有 CTE患者可用的药物治疗，因此任何可能限制或 逆转tau相关的功能障碍将对TBI患者的结果产生重要影响。 此外，鉴于CTE和其他tau蛋白病之间的相似性， 可广泛适用于其他常见的神经退行性疾病。 我们最近开发了一种将机械损伤与tau病理学直接联系起来的体外模型 在培养的神经元中。我们先前研究的一个值得注意的结果是，我们发现突触 功能障碍（使用膜片钳测量）与tau蛋白到树突的错误定位相关 棘（使用荧光成像测量）。这一结果表明， 基于图像的读出可用于筛选药理学试剂对 创伤诱导的tau蛋白病的功能进展。 目前还没有高通量的体外模型来筛选 治疗创伤诱导的tau蛋白病的药物我们已经开发了几种细胞拉伸 系统，用于神经创伤和其他应用。然而，所有这些系统都太 用于进行药物发现研究的低通量。因此，我们的目标是扩大我们的电流， 体外神经创伤模型的药物筛选研究足够高的吞吐量。我们将设计 一种新的用于神经细胞培养的可拉伸多孔板和一种新的高应变率拉伸器 它可以给培养板中的细胞施加类似创伤的负荷。此外，我们将使用机器 基于学习的算法，以快速有效地分析从我们的新系统收集的数据。 设备.最后，我们将使用该设备来测试已知对其他方面有效的抑制剂 神经元损伤影响tau蛋白的错误定位