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，这反过来又是由于使用药物发现平台，而这些平台往往不是 与目标疾病有关。人类诱导性多能干细胞（iPSC）的研究进展 使得在人类神经元上筛选和验证候选化合物成为可能，包括那些来自 患者，从而潜在地提高成功率并加快CNS药物开发的步伐。 BrainXell公司一直是开发基于人类患者神经细胞的CNS平台的先驱 药物发现。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。在第一阶段，我们 确定了几种分子，加速突触素（SYP）的表达，特别是在 皮质谷氨酸能（Glut）神经元或脊髓运动神经元（MN），并配制Glut和MN成熟 这些补充剂在一周内产生表达突触标记的成熟神经元。 在这个第二阶段的项目中，我们建议为其他两个神经元开发成熟补充剂。 亚型，皮质GABA能神经元和中脑多巴胺能神经元。这个项目的首要目标是 一个建议是加速神经元成熟，同时保持高纯度和同质的群体， 神经元，这将允许在HTS中实际使用成熟的人类神经元用于神经元活性和功能。 用于快速产生人成熟神经元的有效鸡尾酒的配制和确定最佳的 它的应用条件将消除建立基于人类患者神经元的HTS的主要障碍 用于中枢神经系统药物的发现。

Project Summary/Abstract Neurological and psychiatric disorders, including drug abuse and addiction, exert a devastating personal and economic toll on patients, families, caregivers, and society. This is in part due to our failure in developing effective medications, which in turn is due to the use of drug discovery platforms that are often not relevant to target diseases. The recent development of induced pluripotent stem cells (iPSCs) from humans makes it possible to screen and validate candidate compounds on human neurons, including those from patients, thus potentially increasing the success rate and speeding up the pace of CNS drug development. BrainXell, Inc. has been a pioneer 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. In our Phase I project, we identified several molecules that accelerate the expression of synaptophysin (SYP) expression specifically in cortical glutamatergic (Glut) neurons or spinal motor neurons (MN), and formulated Glut and MN maturation supplements that yield mature neurons expressing synaptic markers within a week. In this Phase II project, we propose to develop maturation supplements for the other two neuron subtypes, cortical GABAergic neurons and midbrain dopaminergic neurons. The overarching goal of this proposal is to accelerate neuronal maturation while maintaining a highly pure and homogenous population of neurons, which would allow the practical use of mature human neurons in HTS for neuron activity and function. Formulation of an effective cocktail for rapidly generating human mature neurons and determining the optimal conditions for its application will remove the major roadblock in establishing human patient neuron-based HTS for CNS drug discovery.