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

AB 介导的神经元细胞周期折返的精密高内涵筛选测定

• 批准号: 9904312
• 负责人: ELIZABETH SHARLOW
• 金额: $ 40.38万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904312
• 关键词: 3-Dimensional; Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease diagnosis; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Behavioral Symptoms; Biological Assay; Brain; Brain region; Cell Culture Techniques; Cell Cycle; Cell Line; Cell division; Cell model; Chemicals; Coculture Techniques; Cognition; Cyclin D1; Cytokinesis; DNA; Disease Progression; Exposure to; Gene Proteins; Goals; Human; Libraries; Measures; Mediating; Memory; Methodology; Modeling; Molecular; Molecular Target; Mus; Neurons; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Process; Senile Plaques; Signal Pathway; Signal Transduction; Small Molecule Chemical Library; Surrogate Markers; Synapses; System; abeta oligomer; base; high throughput screening; induced pluripotent stem cell; insight; nerve stem cell; neuron loss; novel therapeutics; prevent; relating to nervous system; screening; small molecule; small molecule inhibitor; small molecule libraries

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.

项目摘要 控制记忆和认知的神经元的死亡是导致行为症状的原因。 阿尔茨海默病（AD）。AD中神经元死亡的最常见途径之一是细胞周期重入 (CCR)，其代表神经元细胞周期机制的再激活。通常，分化的神经元 虽然他们从未尝试分裂，但受AD影响的大脑区域中高达5-10%的神经元显示出CCR的迹象。 这些神经元通常复制了大部分DNA，无法进行胞质分裂。而是 最终死亡，并且可能占AD中所见的神经元损失的多达90%。该过程启动 暴露于可溶性淀粉样蛋白-β寡聚体（AβOs），A β Os是不溶性淀粉样蛋白的组成部分 在AD大脑中积累的斑块。鉴定介导Aβ O介导的神经元CCR的基因和蛋白 定义相关的信令网络为早期AD诊断和开发新的AD提供了希望 治疗学我们已经建立了一个Aβ O介导的神经元CCR的人神经细胞模型， 神经iPS细胞系。我们打算利用这些iPS系和衍生的亚系开发高含量的 通过HCS筛选试验鉴定抑制Aβ O介导的CCR的化学型。我们相信，通过这个分析， 我们可以鉴定出可以阻断Aβ O诱导的神经元CCR的小分子，并作为化学探针发挥作用， (1)了解CCR导致神经元死亡的信号通路，以及（2）作为亲本 药物的化学类型。