Novel 3D brain tissue-based screening assay for targeting microglia in CNS neurodegeneration

基于 3D 脑组织的新型筛选方法，用于靶向中枢神经系统神经变性中的小胶质细胞

• 批准号: 9168442
• 负责人: DONALD C LO
• 金额: $ 23.85万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9168442
• 关键词: 3-Dimensional; Address; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein Precursor; Area; Biological Assay; Brain; Cell Line; Cells; Central Nervous System Diseases; Clinical; Corpus striatum structure; Development; Disease; Disease Progression; Drug Targeting; Evaluation; Exons; Frontotemporal Dementia; Glucose; Goals; Heat shock proteins; Huntington Disease; Huntington gene; Immune; Infiltration; Inflammation; Injury; Ischemic Stroke; Knock-out; Microglia; Modeling; Mouse Strains; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Oxygen; Peripheral; Phenotype; Physiological; Process; Protein Isoforms; Public Health; Reporting; Research; Rest; Slice; Staging; Stroke; Therapeutic; Time; Tissue Model; Tissues; Validation; base; brain cell; brain tissue; clinically relevant; cost; deprivation; drug candidate; drug development; drug discovery; in vivo; interest; monocyte; mutant; nervous system disorder; neuroinflammation; novel; novel therapeutics; polyglutamine; protein aggregate; relating to nervous system; research study; screening; stress protein; tau Proteins; three dimensional structure

Aspects of microglial activation in neuroinflammation associated with CNS neurodegeneration have alternately been reported to be further damaging or to be protective against disease progression. Thus, the translational potential of targeting microglia in new drug development for CNS neurodegenerative diseases remains uncertain. A major challenge in this context has been that relevant microglial phenotypes and activation states have been exceedingly difficult to recapitulate in cell line models or even in primary cultures of microglia. Conversely, microglial studies in vivo are time- and cost-intensive, and consequently have limited scalability. To address this need, we propose to develop and provide validation for a novel, brain tissue- based drug discovery model for the identification and mechanistic evaluation of new drug and drug target candidates for modulating microglial activation in CNS neurodegenerative disorders. Brain tissue models capture important aspects of intercellular interactions within the intact, local 3-dimensional structure of native neural tissues, and thereby have increased physiological relevance and can be more predictive of clinical benefit compared to cell-based models. Moreover, we have shown previously that brain slice assays can be scaled to useful throughputs for drug discovery in Huntington's disease (HD), Alzheimer's disease (AD), and stroke. The goal of the present proposal is thus to establish the experimental framework for a brain slice-based screening and mechanistic assay for microglial-neuronal interactions, and to provide initial validation that perturbation of microglial activation and/or numbers leads to clear and reproducible effects on rates and/or extents of neurodegeneration. In addition, we will extend the assay to interrogate potential effects of peripheral monocytes, whose infiltration is associated with later stages of CNS disease. We will initially focus on an HD brain slice model that we have used extensively in both screening as well as mechanistic studies, and then ask if our findings are generalizable to different models of CNS neurodegeneration driven by amyloid precursor protein and tau isoforms relevant to AD and frontotemporal dementias (FTD), respectively. If successful, the proposed studies should provide a new 3-D brain tissue-based model for capturing clinically relevant microglial-neuronal interactions scalable to screening throughputs for the discovery of new candidate drugs and drug targets for CNS neurodegeneration, and for their mechanistic evaluation and validation.

与CNS神经变性相关的神经炎症中小胶质激活的方面 据报道，据报道会进一步破坏或保护疾病 进展。因此，将小胶质细胞靶向新药开发中的翻译潜力 中枢神经系统神经退行性疾病仍然不确定。在这种情况下的主要挑战是 相关的小胶质表型和激活状态非常困难 在细胞系模型甚至小胶质细胞培养物中概括。相反，小胶质细胞 体内的研究是时间和成本密集的，因此可扩展性有限。 为了满足这一需求，我们建议为新颖的脑组织开发和提供验证 基于新药的识别和机理评估的基于药物发现模型 药物目标候选物用于调节中枢神经系统神经退行性疾病中的小胶质细胞激活。 脑组织模型捕获了完整的局部局部间相互作用的重要方面 天然神经组织的3维结构，从而增加了生理 与基于细胞的模型相比，相关性，可以更可预测临床益处。 此外，我们先前已经证明，大脑切片分析可以缩放到有用的吞吐量 用于亨廷顿氏病（HD），阿尔茨海默氏病（AD）和中风的药物发现。 因此，本提案的目的是为大脑建立实验框架 基于切片的筛选和小胶质细胞神经元相互作用的机理测定，并提供 初始验证小胶质激活和/或数字的扰动导致清除和 对神经变性的速率和/或范围的可再现影响。此外，我们将扩展 测定询问周围单核细胞的潜在影响，其浸润与 CNS病的后期阶段。我们最初将专注于我们使用过的高清脑切片模型 在筛选和机械研究中广泛，然后询问我们的发现是否是 可推广到由淀粉样前体蛋白驱动的CNS神经退行性模型的不同模型 和与AD和额颞痴呆（FTD）相关的TAU同工型。 如果成功，拟议的研究应为新的3-D脑组织模型提供 捕获可扩展到可扩展到筛查吞吐量的临床相关的小胶质细胞 - 神经元相互作用 发现新的候选药物和中枢神经系统神经退行性的药物靶标的 机械评估和验证。