Bioartificial Brain Slices for Drug Screening

描述（由申请人提供）：ISurTec正在寻求NIH的资助，以开发一种人类生物人工脑切片原型，用于电生理药物筛选试验。从大鼠或小鼠制备的急性脑切片目前是表征神经组织对药物、毒素或其他药剂的反应的“金标准”制备。然而，每只动物产生的脑切片数量有限，加上动物护理和使用的高成本，仍然是这种方法的主要限制。此外，由于种间遗传变异，通过使用急性脑切片获得的临床前数据通常对人体临床试验和上市后监测的预测能力有限。在这项研究中，三维神经回路支架将被图案化后的谢弗侧支-海马CA 1区锥体细胞突触。350微米厚的支架将通过层压和退火无规非织造材料和对齐的光反应性增透垫，然后对其进行表面改性来制备。所得的生物人工脑切片支架将为共培养的神经元群体建立开放场电路架构，以响应于顺向刺激产生突触后场电位。在第一阶段，将制备原型支架，并用E18大鼠海马神经元填充，以确认支架支持产生强大的突触后场电位的能力。I阶段II，支架将用源自可扩增神经祖细胞（hNP）细胞系的人神经元填充。在生长因子图案的支架上原位的hNPs的末端分化也将在第二阶段进行研究。生物人工脑切片支架将以标准化的形式生产，与常见的电生理设备兼容。最终，我们设想支架将使电生理检测具有高预测能力的多种神经递质系统，包括多巴胺能，胆碱能，多巴胺能和gabaergic突触。因此，人类生物人工脑切片将是药物开发的有价值的工具，涉及各种人类神经系统疾病和障碍，包括成瘾，ADD/ADHD，抑郁症，帕金森病，疼痛处理和精神分裂症。该技术将通过克服来自啮齿动物的急性切片的两个局限性来推进HTS领域（1）有限的人类相关性和（2）动物护理合规性和监督的高负担和成本。

DESCRIPTION (provided by applicant): ISurTec is seeking NIH funding to develop a human bioartificial brain slice prototype, designed for use in electrophysiological drug-screening assays. Acute brain slices, prepared from rats or mice, are currently the 'gold standard' preparation for characterizing neural tissue responses to drugs, toxins, or other agents. However, the limited number of brain slices produced per animal, coupled with the high costs of animal care and use, remains a major limitation of this approach. Furthermore, due to interspecies genetic variation, preclinical data acquired through the use of acute brain slices wil usually be of limited predictive power for human clinical trials and postmarket surveillance. In this study, 3-dimensional neural circuit scaffolds will be patterned after the Schaffer collateral-CA1 pyramidal cell synapse of the hippocampus. 350 micron thick scaffolds will be prepared by laminating and annealing random nonwoven and aligned photoreactive nanofiber mats, following their photopatterned surface modification. The resulting bioartificial brain slice scaffolds will establish open field circuit architecture for co- cultured neuronal populations to generate postsynaptic field potentials in response to orthodromic stimulation. In Phase I, prototype scaffolds will be prepared and populated with E18 rat hippocampal neurons, to confirm the scaffold's ability to support the generation of robust postsynaptic field potentials. I Phase II, scaffolds will be populated with human neurons derived from expandable Neural Progenitor (hNP) cell lines. Terminal differentiation of hNPs in situ on growth factor-patterned scaffolds will also be investigated in Phase II. The bioartificial brain slice scaffold will be produced in a standardized format that is compatible with common electrophysiological equipment. Ultimately, we envision the scaffolds will enable electrophysiological assays with high predictive power for multiple neurotransmitter systems including glutamatergic, cholinergic, dopaminergic, and gabaergic synapses. Thus, the human bioartificial brain slice will be a valuable tool for drug development concerning a wide variety of human neurological diseases and disorders, including addiction, ADD/ADHD, depression, Parkinson's disease, pain processing, and schizophrenia. This technology will advance the field of HTS by overcoming two limitations of acute slices derived from rodents (1) limited human relevance and (2) high burden and cost of animal care compliance and oversight.