Accelerating Functional Maturation of Human iPSC-Derived Astrocytes

项目摘要/摘要 神经和精神障碍，包括阿尔茨海默病，对个人和 患者、家庭、照顾者和社会的经济损失。这在一定程度上是因为我们未能开发出有效的 药物，反映了药物发现平台往往与目标疾病无关。最近的 人类诱导多能干细胞(IPSCs)的开发使筛选和验证成为可能 人类脑细胞上的候选化合物，包括来自患者的那些，因此潜在地增加了 成功率和加快中枢神经系统药物开发的步伐。 BrainXell，Inc.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 发现号。星形胶质细胞的功能丧失或损害与广泛的病理过程有关 以及神经障碍，包括阿尔茨海默病、亨廷顿病和肌萎缩侧索硬化症。 我们开发了一种新的方法，可以从IPSCs中快速生成丰富的具有功能的星形胶质细胞 Week通过应用胶质形成转录因子NFIA和SOX9的可诱导表达。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，这使得它很难呈现。 用于疾病表型和高通量筛选(HTS)，以寻找针对大脑疾病患者的药物线索 是完全成熟的。这个第一阶段SBIR项目的目标是发现能够加速 IPSC来源的星形胶质细胞中与成熟状态相关的基因和产生成熟的鸡尾酒 培养后1-3周内可见星形胶质细胞。我们将设计一种带有纳米纤维素酶(Nluc)的人类iPSC报道线 融合到EAAT2(由SLC1A2基因编码)，一个在成熟星形胶质细胞中高表达的谷氨酸转运体。 这条线将使筛选促进星形胶质细胞成熟的小分子成为可能并简化这一过程。 快速生成成熟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 在建立以人星形胶质细胞为基础的HTS用于中枢神经系统药物开发方面。

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.