Automation of Assay Endpoints for Brain Slice Models of Neurodegenerative Disease

描述（由申请人提供）：对于神经和神经精神疾病的新药和药物靶点发现，从基于细胞的试验的有效性过渡到在全动物模型中的获益一直是困难和不确定的。理想情况下，药物发现研究应该尽可能地在全动物疾病模型中进行，但在动物体内进行实验是非常昂贵和耗时的。相反，尽管基于细胞系和原代培养的检测快速且廉价，但当神经元培养和/或永生化时，它们会受到表型变化的影响，更重要的是，正常的三维环境和脑组织结构的局部细胞间相互作用不可避免地会丢失。为了帮助弥合基于细胞和全动物功效研究之间的差距，我们开发了一系列基于完整脑组织的中枢神经系统疾病模型，包括中风、亨廷顿病（HD）和阿尔茨海默病（AD）。在我们发表的研究中，我们已经展示了这种检测在推进一系列基因靶标鉴定和药物开发计划中的效用。为了支持这些脑切片外植体模型在大规模发现工作中的使用，我们在过去十年中开发了许多技术和工艺创新，包括高通量脑切片和用于转染疾病相关基因的脑切片的生物基因枪装置

DESCRIPTION (provided by applicant): For new drug and drug target discovery in neurological and neuropsychiatric disorders, transitioning from efficacy in cell-based assays to benefit in whole-animal models has always been difficult and uncertain. Ideally, drug discovery studies would be conducted, as much as possible, in whole-animal models of disease, but in vivo animal experiments are tremendously costly and time- consuming. Conversely, while cell line and primary culture-based assays are rapid and inexpensive, they are compromised by phenotypic changes when neurons are cultured and/or immortalized, and, importantly, by the normal 3-dimensional milieu and local inter-cellular interactions of brain tissue architecture being unavoidably lost. To help bridge this gap between cell-based and whole-animal efficacy studies, we have developed a series of intact brain tissue-based models for CNS disorders including stroke, Huntington's disease (HD), and Alzheimer's disease (AD). In our published studies, we have shown the utility of such assays in advancing a range of gene target identification and drug development programs. To support the use of these brain slice explant models in the context of larger-scale discovery efforts, we have developed numerous technological and process innovations over the last decade, including high-throughput brain slicing and biolistic gene gun devices for transfection of brain slices with disease-relevant genes and assay reporter constructs. The overarching goal of the present proposal is to solve the final rate-limiting barrier to full scalability of this approach, namely, the automation of brain slice-based assay endpoints. To date, all of the brain slice disease models we have developed have been analyzed using laborious manual endpoint assays; nevertheless, assay throughput has been sufficient to support the screening of hundreds to thousands of compounds or gene targets per year even with a modest-sized scientific team. Implementation of "turnkey" unbiased, automated microscopy and high-content analysis (HCA) platforms would increase the throughput of these assays by 10-fold or more, and enable full support of large-scale systems biology, bioinformatics, and drug discovery programs. Such a bridging stage of "high-throughput biology" screening between cell-based and whole-animal models should significantly increase the likelihood of success of drug and drug target discovery and development programs, by providing a preview of both efficacy as well as potential adverse off-target effects in intact neurl tissue assays before substantial time and financial commitments are made to full in vivo efficacy studies.