Accelerating Functional Maturation of Human iPSC-Derived Microglia

项目总结/摘要 神经和精神疾病，包括阿尔茨海默氏病（AD），对个人造成毁灭性的影响。 以及对患者、家庭、护理人员和社会造成的经济损失。这部分是由于我们未能发展 有效的药物，反映了药物发现平台往往与目标疾病无关。的 来自人类的诱导多能干细胞（iPSC）的最新发展使得筛选和 在人脑细胞上验证候选化合物，包括来自患者的化合物，从而可能增加 成功率和加快CNS药物开发的步伐。 BrainXell公司率先开发了基于人类患者脑细胞的中枢神经系统药物平台， 的发现我们能够从人类中生产大量质量一致的高度富集的小胶质细胞， iPSC通过我们的平台技术定向分化和扩增定向祖细胞。 然而，iPSC衍生的小胶质细胞是不成熟的，可与胎儿阶段的小胶质细胞相比，这使得很难 用于呈现疾病表型和用于高通量筛选（HTS）预期用于 那些大脑完全成熟的人第一阶段SBIR项目的目标是发现能够加速 与iPSC衍生的小胶质细胞中成熟状态相关的基因的表达，并配制 在接种后1-3周内产生成熟的小胶质细胞。我们将设计一条人类iPSC报告线， 纳米荧光素酶（Nluc）与P2 RY 12融合，P2 RY 12是在成熟小胶质细胞中高度表达的嘌呤能受体。这条线路将 能够并简化用于加速小胶质细胞成熟的小分子的筛选。制订 用于快速产生成熟人类小胶质细胞的有效鸡尾酒将消除建立一种新的神经细胞的主要障碍， 用于CNS药物开发基于人类患者小胶质细胞的HTS。

Project Summary/Abstract Neurological and psychiatric disorders, including Alzheimer’s disease (AD), exert a devastating personal and economic toll on patients, families, caregivers, and society. This is in part due to our failure to develop effective medications, reflecting 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 brain cells, including those from patients, thus potentially increasing the success rate and speeding the pace of CNS drug development. BrainXell, Inc. has pioneered the development of human patient brain cell-based platforms for CNS drug discovery. We are able to produce large quantities of highly enriched microglia of consistent quality from human iPSCs through our platform technology of directed differentiation and expansion of committed progenitors. However, iPSC-derived microglia 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 speed the expression of genes associated with a mature state in iPSC-derived microglia and to formulate cocktails that yield mature microglia within 1-3 weeks after plating. We will engineer a human iPSC reporter line with nanoluciferase (Nluc) fused to P2RY12, a purinergic receptor highly expressed in mature microglia. This line will enable and simplify the screening of small molecules for accelerating microglia maturation. Formulation of an effective cocktail for rapidly generating mature human microglia will remove a major roadblock in establishing human patient microglia-based HTS for CNS drug development.