High-Throughput In Vitro Analyses of Trauma-Induced Tauopathy

项目总结 慢性创伤性脑病(CTE)是一种与 反复轻度颅脑损伤(TBI)。CTE是许多神经退行性疾病中的一种 特征为tauopathy，其中tau蛋白通常与 神经元轴突中的微管与微管分离，启动了 退行性级联反应，最终导致神经功能丧失。目前没有 为CTE患者提供的药物治疗，因此任何可能限制或 逆转tau相关功能障碍将对脑外伤患者的预后产生重要影响。 此外，鉴于CTE与其他疾病之间的相似之处，对CTE治疗的见解 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体外模型，将机械损伤与tau病理直接联系起来。 在培养的神经元中。我们之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向树突状细胞的错误定位有关 脊椎(使用荧光成像测量)。这一结果表明，这一相对简单的 基于图像的读数可用于筛选药理药物对心脏功能的影响。 外伤性直肠痉挛的功能进展。 目前还没有高通量的体外模型来筛选 治疗创伤引起的紧张症的药物。我们已经开发了几种细胞伸展 系统，既用于神经创伤，也用于其他应用。然而，所有这些系统都太 进行药物发现研究的吞吐量低。因此，我们的目标是扩大我们目前的 体外神经创伤模型要有足够高的吞吐量进行药物筛选研究。我们将设计 一种用于神经细胞培养的新型可伸缩多孔板和新型高应变率拉伸器 可以对平板中的细胞施加创伤般的负载。此外，我们还将使用机器 基于学习的算法，以快速高效地分析从我们的新 设备。最后，我们将使用该设备测试已知对其他方面有效的抑制剂 神经元损伤对tau错误定位的影响

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