Large-Scale Production of High Purity Human Neurons for CNS Drug Discovery

 描述（由申请人提供）：中枢神经系统（CNS）疾病给美国带来的个人、社会和经济负担是巨大的。仅精神保健一项，每年的直接费用估计为575亿美元。这部分是由于未能开发有效的药物。从2003年到2014年，FDA只批准了37种治疗CNS疾病的新药，涵盖神经病学，精神病学和疼痛的临床领域。这平均每年只有三个新的化学实体（NEI），而CNS药物开发的临床试验失败率仍超过90%。尽管这种极高的损耗水平的原因是无数的和复杂的，药物开发的核心挑战是，用于早期临床前鉴定和验证新化合物的平台通常与目标疾病无关。 BrainXell公司试图通过产生更接近人类CNS的体外模型系统来解决这些问题。SBIR第一阶段的建议是生产大量的高度富集，功能特异性神经元的一致质量，满足高通量筛选（HTS）的要求。我们选择了皮层神经元作为这个项目的神经元亚型。这些神经元在许多精神疾病中受到影响，包括抑郁症和精神分裂症。我们已经设计了一种具有纳米荧光素酶组成型表达的人iPSC系，使我们能够评估旨在快速扩增皮质神经祖细胞的小分子组合。这一目标将通过修改以前用于产生大量高纯度运动神经元的方法来实现。我们还将确定和开发新的解离溶液和冷冻介质，以促进解聚和解冻后的存活，从而降低成本，使其适用于HTS。制定一个有效的鸡尾酒扩大和维持皮质祖细胞不仅使我们能够 本发明的方法可以从正常以及患者iPSC制造用于HTS的皮质GABA能神经元，而且还提供了用其他神经元亚型（例如皮质GABA能神经元或中脑多巴胺能神经元）实现相同目标的技术基础。

 DESCRIPTION (provided by applicant): The personal, societal, and economic burden that disorders of the central nervous system (CNS) place on the United States is tremendous. For mental health care alone, direct costs were estimated at $57.5 billion annually. This is in part due to failures in developing effective medications. From 2003-2014, the FDA approved just 37 new drugs to treat CNS disorders covering the clinical areas of neurology, psychiatry, and pain. That averages to just three new chemical entities (NEIs) per year, while clinical trial failure rats for CNS drug development remain greater than 90%. Although the reasons for this exceedingly high level of attrition are myriad and complex, the central challenge to drug development is that platforms for early preclinical identification and validation of novel compounds are often not relevant to the target disease. BrainXell, Inc. seeks to address these issues head-on by generating in vitro model systems that more closely approximate the human CNS. This SBIR Phase I proposal is to produce large quantities of highly enriched, functionally specialized neurons of consistent quality that meet high-throughput screening (HTS) requirements. We have selected cortical glutamatergic neurons as the neuron subtype for this project. These neurons are affected in a large number of psychiatric diseases, including depression and schizophrenia. We have engineered a human iPSC line with constitutive expression of nanoluciferase, allowing us to evaluate combinations of small molecules designed to rapidly expand cortical neural progenitors. This goal will be achieved by modifying approaches used previously to generate large numbers of highly pure motor neurons. We will also identify and develop novel dissociation solutions and freezing media to promote survival following disaggregation and thawing, thus reducing costs and making it practical for HTS. Formulation of an effective cocktail for expanding and maintaining cortical progenitors will not only enable us to manufacture cortical glutamatergic neurons for HTS from normal as well as patient iPSCs, but also provide the technical basis to accomplish the same goal with other neuronal subtypes, such as cortical GABAergic neurons or midbrain dopaminergic neurons.