Accelerating Functional Maturation of Human iPSC-Derived Astrocytes

项目总结/摘要 神经和精神疾病，包括阿尔茨海默氏病，对个人和社会造成毁灭性的影响。 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有效地开发 这反映了药物发现平台往往与目标疾病无关。近期 人类诱导多能干细胞（iPSC）的开发使得筛选和验证 候选化合物对人类脑细胞的作用，包括来自患者的那些，从而潜在地增加 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， 的发现星形胶质细胞的功能丧失或损伤与广泛的病理过程有关 和神经障碍，包括阿尔茨海默病、亨廷顿病和肌萎缩侧索硬化。 我们开发了一种新的方法，用于在四个实验室中从iPSC快速产生富集和功能性星形胶质细胞。 周通过应用胶质生成转录因子NFIA和SOX 9的诱导表达。然而，iPSC- 衍生的星形胶质细胞是不成熟的，与胎儿阶段的星形胶质细胞相当，这使得其难以呈现。 疾病表型和高通量筛选（HTS）的药物线索，旨在为那些 完全成熟了。SBIR项目第一阶段的目标是发现加速表达的分子， 与iPSC衍生的星形胶质细胞中的成熟状态相关的基因，并配制产生成熟的 接种后1-3周内的星形胶质细胞。我们将用纳米荧光素酶（Nluc）工程化人类iPSC报告细胞系。 融合到EAAT 2（由SLC 1A 2基因编码），一种在成熟星形胶质细胞中高度表达的谷氨酸转运蛋白。 这条生产线将使加速星形胶质细胞成熟的小分子筛选变得可能和简单。 快速生成成熟的人类星形胶质细胞的有效鸡尾酒配方将消除一个主要障碍 在建立用于CNS药物开发的基于人类患者星形胶质细胞的HTS方面。

Project Summary/Abstract Neurological and psychiatric disorders, including Alzheimer’s disease, 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. Functional loss or impairment of astrocytes is implicated in a wide range of pathological processes and neural disorders, including Alzheimer's disease, Huntington’s disease, and amyotrophic lateral sclerosis. We developed a novel method for rapid generation of enriched and functional astrocytes from iPSCs in four weeks by applying inducible expression of gliogenic transcription factors NFIA and SOX9. However, iPSC- derived astrocytes 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 astrocyte and to formulate cocktails that yield mature astrocyte within 1-3 weeks after plating. We will engineer a human iPSC reporter line with nanoluciferase (Nluc) fused to EAAT2 (encoded by the SLC1A2 gene), a glutamate transporter highly expressed in mature astrocytes. This line will enable and simplify the screening of small molecules for accelerating astrocyte maturation. Formulation of an effective cocktail for rapidly generating mature human astrocyte will remove a major roadblock in establishing human patient astrocyte-based HTS for CNS drug development.