Accelerating Functional Maturation of Human Neurons for High-Throughput Screening

项目摘要/摘要 神经和精神障碍，包括药物滥用和成瘾，会造成毁灭性的 患者、家属、照顾者和社会的个人和经济损失。这在一定程度上是因为我们在 开发有效的药物，这些药物反过来又流向药物发现平台，而这些平台往往与 针对疾病。最近人类诱导多能干细胞(IPSCs)的发展使这成为可能 筛选和验证人类神经细胞上的候选化合物，包括来自患者的化合物，从而 潜在地提高了中枢神经系统药物开发的成功率并加快了步伐。 BrainXell，Inc.率先为中枢神经系统药物开发基于人类患者神经细胞的平台 发现号。我们能够产生大量高度丰富的、功能专门化的神经元 通过我们的定向神经分化平台技术从人类IPSCs获得稳定的质量 扩大承诺的祖辈。然而，IPSC来源的神经元是不成熟的，与 胎儿期，这使得疾病表型的表现和高通量很难 筛选(HTS)针对大脑完全成熟的人的药物线索。这一阶段SBIR的目标是 项目是发现加速皮质谷氨酸能和中脑成熟的分子 多巴胺能神经元，并配制出在培养一周内产生成熟神经元的鸡尾酒 祖先。我们已经设计出了一个人ipSC报道系，它融合了纳米荧光素酶(Nluc)和 突触素(SYP)，一种几乎在所有成熟神经元中表达的突触囊泡糖蛋白，使 简化小分子的筛选，以加速神经元成熟。制定一项有效的 用于快速生成人类成熟神经元的鸡尾酒将消除建立人类 基于患者神经元的HTS用于中枢神经系统药物开发。

Project Summary/Abstract Neurological and psychiatric disorders, including substance abuse and addiction, exert a devastating personal and economic toll on patients, families, caregivers, and society. This is in part due to our failures in developing effective medications, which in turn to the drug discovery platforms that are often not relevant to target diseases. Recent development of induced pluripotent stem cells (iPSCs) from humans makes it possible to screen and validate candidate compounds on human nerve cells, including those from patients, thus potentially increasing the success rate and speeding up the pace in CNS drug development. BrainXell, Inc. has pioneered in developing human patient neural cell-based platforms for CNS drug discovery. We are able to produce large quantities of highly enriched, functionally specialized neurons of consistent quality from human iPSCs through our platform technology of directed neural differentiation and expansion of committed progenitors. However, iPSC-derived neurons are immature, comparable to those at the fetal stage, which makes it difficult for presentation of disease phenotypes and for high-throughput screening (HTS) for drug leads intended for those whose brains are fully mature. The goal of this Phase I SBIR project is to uncover molecules that accelerate the maturation of cortical glutamatergic and midbrain dopaminergic neurons and to formulate cocktails that yield mature neurons within a week of plating their progenitors. We have engineered a human iPSC reporter line with the fusion of nanoluciferase (Nluc) with synaptophysin (SYP), a synaptic vesicle glycoprotein expressed in virtually all mature neurons to enable and simplify the screening of small molecules for accelerating neuronal maturation. Formulation of an effective cocktail for rapidly generating human mature neurons will remove the major roadblock in establishing human patient neuron-based HTS for CNS drug development.