Continuous Monitoring of Tau-Induced Neurotoxicity in Patient-Derived iPSC-Neurons.
Continuous Monitoring of Tau-Induced Neurotoxicity in Patient-Derived iPSC-Neurons.
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DOI：
10.1523/jneurosci.2590-20.2021
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发表时间：
2021-05-12
期刊：
影响因子：
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通讯作者：
Frosch MP
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文献类型：
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作者：
Oakley DH;Klickstein N;Commins C;Chung M;Dujardin S;Bennett RE;Hyman BT;Frosch MP
神经元内的Tau聚集是阿尔茨海默病（AD）和相关Tau病变的关键特征。据信，可溶性病理性tau种类以朊病毒样方式播种tau聚集体的形成，并在疾病进展期间通过连接的神经元传播。可溶性和聚集形式的tau被认为具有神经毒性性质。此外，不同株的错折叠tau蛋白可能导致不同的神经毒性。在这项工作中，我们提出了一个加速人类神经元模型的tau诱导的神经毒性，结合可溶性tau物种和tau聚集。使用表达tau聚集生物传感器的患者来源的诱导多能干细胞（iPSC）神经元，我们开发了一种细胞培养系统，其允许以单细胞分辨率连续评估诱导的tau聚集和神经元活力，持续时间>1周。我们表明，外源性tau“种子”的摄取，测量tau重复结构域（TauRD）报告聚集，增加了随后的神经元死亡的风险在体外。这些结果首次直接显现单个人iPSC神经元中的神经元TauRD聚集和随后的细胞死亡。然后鉴定特定的形态菌株或TauRD聚集模式，并与不同的神经毒性相关。此外，我们证明了表达PSEN 1 L435 F突变的家族性AD iPSC神经元与对照iPSC神经元相比表现出加速的TauRD聚集动力学和tau菌株传播偏倚。微管结合蛋白tau的神经元细胞内聚集发生在阿尔茨海默病和相关的神经退行性tau蛋白病中。Tau聚集体被认为通过朊病毒样错误折叠的Tau种子从神经元扩散到神经元。我们的工作开发了一种人类神经元实时成像系统，以可视化单个神经元内的种子tau聚集和tau诱导的神经毒性。使用聚集感应tau报告基因，我们发现神经元对tau种子的摄取和繁殖降低了随后的存活率。此外，在早老素-1中携带导致阿尔茨海默病的突变的人诱导多能干细胞（iPSC）神经元比对照iPSC神经元更快地经历tau接种。然而，它们在神经元存活方面没有表现出随后的差异。最后，tau聚集体的特定形态与增加的神经毒性相关。
Tau aggregation within neurons is a critical feature of Alzheimer's disease (AD) and related tauopathies. It is believed that soluble pathologic tau species seed the formation of tau aggregates in a prion-like manner and propagate through connected neurons during the progression of disease. Both soluble and aggregated forms of tau are thought to have neurotoxic properties. In addition, different strains of misfolded tau may cause differential neurotoxicity. In this work, we present an accelerated human neuronal model of tau-induced neurotoxicity that incorporates both soluble tau species and tau aggregation. Using patient-derived induced pluripotent stem cell (iPSC) neurons expressing a tau aggregation biosensor, we develop a cell culture system that allows continuous assessment of both induced tau aggregation and neuronal viability at single-cell resolution for periods of >1 week. We show that exogenous tau “seed” uptake, as measured by tau repeat domain (TauRD) reporter aggregation, increases the risk for subsequent neuronal death in vitro. These results are the first to directly visualize neuronal TauRD aggregation and subsequent cell death in single human iPSC neurons. Specific morphologic strains or patterns of TauRD aggregation are then identified and associated with differing neurotoxicity. Furthermore, we demonstrate that familial AD iPSC neurons expressing the PSEN1 L435F mutation exhibit accelerated TauRD aggregation kinetics and a tau strain propagation bias when compared with control iPSC neurons. SIGNIFICANCE STATEMENT Neuronal intracellular aggregation of the microtubule binding protein tau occurs in Alzheimer's disease and related neurodegenerative tauopathies. Tau aggregates are believed to spread from neuron to neuron via prion-like misfolded tau seeds. Our work develops a human neuronal live-imaging system to visualize seeded tau aggregation and tau-induced neurotoxicity within single neurons. Using an aggregation-sensing tau reporter, we find that neuronal uptake and propagation of tau seeds reduces subsequent survival. In addition, human induced pluripotent stem cell (iPSC) neurons carrying an Alzheimer's disease-causing mutation in presenilin-1 undergo tau seeding more rapidly than control iPSC neurons. However, they do not show subsequent differences in neuronal survival. Finally, specific morphologies of tau aggregates are associated with increased neurotoxicity.