In vitro modelling of Alzheimer's disease: Degeneration and cell death induced by viral delivery of amyloid and tau
In vitro modelling of Alzheimer's disease: Degeneration and cell death induced by viral delivery of amyloid and tau
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DOI：
10.1016/j.expneurol.2011.01.018
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发表时间：
2011-06
影响因子：
5.3
通讯作者：
S. Stoppelkamp;H. Bell;Jon Palacios-Filardo;D. Shewan;G. Riedel;B. Platt
中科院分区：
文献类型：
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作者：
S. Stoppelkamp;H. Bell;Jon Palacios-Filardo;D. Shewan;G. Riedel;B. Platt
随着预期寿命的增加，阿尔茨海默病（AD）和其他痴呆症构成了日益严重且尚未解决的健康问题。AD的各种细胞模型有助于破译淀粉样蛋白和tau蛋白相关变性的一些关键方面。合成肽的细胞外应用的最初方法现在已经被淀粉样前体蛋白（APP）和tau基因的引入所取代。在本研究中，利用腺病毒转导将基因递送到原代大鼠海马和背根神经节（DRG）培养物中，以在细胞水平上进行比较和机制研究，并随后进行药物测试。时间推移实验揭示了不同的细胞死亡模式：APP呈类凋亡，而tau阳性细胞连接并形成簇。突变的人APP或tau表达引起加速的神经元损伤和细胞死亡（参见图11）。EGFP：APP在第5天为− 50%; tau在第3天为− 40%）。这种活力的降低之前，降低兴奋性，通过响应去极化KCl的挑战，在Ca 2+成像实验监测。此外，两种转基因都减少了DRG神经元中的神经突生长。治疗研究证实，APP诱导的损伤可以通过β-和γ-分泌酶抑制剂（提供对照水平的60-100%的保护）、氯碘羟喹（80%）和锂（100%）改善;而抗聚集治疗有益于tau诱导的损伤（相对于对照60-90%恢复）。有趣的是，咖啡因是最有希望的候选药物，在APP（77%）和tau诱导的模型（72%的恢复率）中具有高疗效。总的来说，这些细胞模型为AD和相关疾病的机制研究和靶点鉴定提供了优势。
With increasing life expectancy, Alzheimer's disease (AD) and other dementias pose an increasing and as yet unresolved health problem. A variety of cellular models of AD has helped to decipher some key aspects of amyloid and tau related degeneration. The initial approach of extracellular applications of synthetic peptides has now been replaced by the introduction of amyloid precursor protein (APP) and tau genes. In the present study adenoviral transductions were exploited for gene delivery into primary rat hippocampal and dorsal root ganglion (DRG) cultures to enable comparative and mechanistic studies at the cellular level and subsequent drug testing. Time lapse experiments revealed a different pattern of cell death: apoptotic-like for APP whereas tau positive cells joined and formed clusters. Mutated human APP or tau expression caused accelerated neuronal damage and cell death (cf. EGFP: − 50% for APP at 5 days; − 40% for tau at 3 days). This reduction in viability was preceded by decreased excitability, monitored via responses to depolarising KCl-challenges in Ca2+imaging experiments. Additionally, both transgenes reduced neurite outgrowth in DRG neurones. Treatment studies confirmed that APP induced-damage can be ameliorated by β- and γ-secretase inhibitors (providing protection to 60–100% of control levels), clioquinol (80%) and lithium (100%); while anti-aggregation treatments were beneficial for tau-induced damage (60–90% recovery towards controls). Interestingly, caffeine was the most promising drug candidate for therapeutic intervention with high efficacy in both APP (77%) and tau-induced models (72% recovery). Overall, these cellular models offer advantages for mechanistic studies and target identification in AD and related disorders.