Accelerating Functional Maturation of Human Neurons for High-Throughput Screening

项目总结/摘要 神经和精神疾病，包括药物滥用和成瘾， 对患者、家庭、护理人员和社会造成的个人和经济损失。这部分是由于我们的失败， 开发有效的药物，这反过来又与药物发现平台，往往是不相关的， 目标疾病。最近开发的人类诱导多能干细胞（iPSC）使其成为可能， 在人类神经细胞上筛选和验证候选化合物，包括来自患者的化合物， 潜在地增加了成功率并加快了CNS药物开发的步伐。 BrainXell公司在开发基于人类患者神经细胞的CNS药物平台方面处于领先地位 的发现我们能够产生大量高度富集的，功能特化的神经元， 通过我们的定向神经分化平台技术， 扩展承诺的祖细胞。然而，iPSC衍生的神经元是不成熟的，与那些在 胎儿期，这使得疾病表型的呈现和高通量的 药物筛选（HTS）旨在为那些大脑完全成熟的人提供药物。第一阶段SBIR的目标是 一个项目是发现加速大脑皮层神经元和中脑成熟的分子， 多巴胺能神经元，并制定鸡尾酒，产生成熟的神经元在一个星期内铺板， 祖先我们已经工程化了人iPSC报告细胞系，其具有纳米荧光素酶（Nluc）与 突触素（SYP）是一种突触囊泡糖蛋白，在几乎所有成熟神经元中表达， 简化用于加速神经元成熟的小分子的筛选。制定有效的 快速产生人类成熟神经元的鸡尾酒将消除建立人类神经元的主要障碍。 用于CNS药物开发的基于患者神经元的HTS。

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.