Bioartificial Brain Slices for Drug Screening

描述（由申请人提供）：ISurTec正在寻求NIH资助开发人类生物人工脑切片原型，设计用于电生理药物筛选分析。从大鼠或小鼠身上制备的急性脑切片是目前表征神经组织对药物、毒素或其他药物反应的“金标准”制备方法。然而，每只动物的大脑切片数量有限，加上动物护理和使用的高成本，仍然是这种方法的主要局限性。此外，由于物种间的遗传变异，通过使用急性脑切片获得的临床前数据通常对人类临床试验和上市后监测的预测能力有限。在本研究中，三维神经回路支架将以海马的Schaffer侧侧- ca1锥体细胞突触为模板。将随机无纺布和排列的光反应纳米纤维垫层合退火，并对其表面进行光化修饰，制备350微米厚的支架。由此产生的生物人工脑切片支架将为共培养的神经元群建立开放场电路结构，以响应正交刺激产生突触后场电位。在第一阶段，将制备原型支架并填充E18大鼠海马神经元，以证实支架支持产生强大的突触后场电位的能力。在II期，支架将填充来自可扩展神经祖细胞（hNP）细胞系的人类神经元。hNPs在生长因子支架上的原位分化也将在II期进行研究。生物人工脑切片支架将以与普通电生理设备兼容的标准化格式生产。最终，我们设想这种支架将使电生理检测对多种神经递质系统具有高预测能力，包括谷氨酸能、胆碱能、多巴胺能和gabaergy突触。因此，人类生物人工脑切片将是一个有价值的药物开发工具，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(1)有限的人类相关性和(2)高负担和高成本的动物护理依从性和监督，推动HTS领域的发展。

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.