A Precision high content screening assay for AB-mediated neuronal cell cycle reentry

项目摘要 控制记忆和认知的神经元的死亡是导致 阿尔茨海默病(AD)。阿尔茨海默病最常见的神经元死亡途径之一是细胞周期重入 (Ccr)，代表神经细胞周期机制的重新激活。通常，分化的神经元 永远不要尝试分裂，然而，在受AD影响的脑区，高达5%-10%的神经元显示出CCR的迹象。 这些神经元通常复制了大量的DNA，无法进行胞质分裂。相反，他们 最终死亡，可能占阿尔茨海默病神经元丢失的90%。此过程启动 暴露在可溶性淀粉样蛋白-β寡聚体(AβOs)中，这是不溶性淀粉样蛋白的构建块 阿尔茨海默病脑内积聚的斑块。鉴定介导βO介导的神经元CCR的基因和蛋白 定义相关的信令网络为AD的早期诊断和开发新的AD提供了希望 治疗学。我们已经建立了βO介导的神经细胞CCR的人类神经细胞模型，该模型利用 神经性iPS细胞系。我们打算利用这些iPS系和衍生的亚系来开发高含量 筛选抑制AβO介导的CCR的化学类型。我们相信，通过这个化验， 我们可以识别出可以阻断βO诱导的神经元CCR的小分子，并发挥化学探针的作用 (1)了解CCR导致神经元死亡的信号通路；(2)作为亲本 药物的化学类型。

Project Summary The death of neurons that control memory and cognition are responsible for the behavioral symptoms of Alzheimer’s disease (AD). One of the most common pathways for neuronal death in AD is cell cycle re-entry (CCR), which represents the reactivation of neuronal cell cycle machinery. Usually, differentiated neurons never attempt to divide, yet up to 5-10% of the neurons in brain regions affected by AD show signs of CCR. These neurons, which typically have duplicated much of their DNA, fail to undergo cytokinesis. Instead, they eventually die and may account for as much as 90% of the neuronal loss seen in AD. This process initiates with exposure to soluble amyloid-β oligomers (AβOs), which are the building blocks of the insoluble amyloid plaques that accumulate in AD brain. Identifying genes and proteins that mediate AβO-mediated neuronal CCR and defining relevant signaling networks hold promise for early AD diagnosis and for developing new AD therapeutics. We have developed a human neural cell model of AβO-mediated neuronal CCR, which utilizes neural iPS cell lines. We intend to use these iPS lines and derived sublines to develop a high content screening (HCS) assay to identify chemotypes that inhibit AβO-mediated CCR. We believe, through this assay, we can identify small molecules that can block AβO-induced neuronal CCR and function as chemical probes to (1) understand the signaling pathways leading to neuronal death by CCR and (2) serve as parental chemotypes for drugs.