Alzheimer's Disease Risk Genes in Human Microglia and Neurons Derived from iPSCs

描述（由申请人提供）：到2050年，1380万美国人患有阿尔茨海默病（AD）的幽灵迫在眉睫，这促使我们在典型的“细胞系到啮齿动物到人体试验”模型之外扩展我们的生物医学研究范式。迄今为止，所有进入III期临床试验的AD药物的表现令人失望，这也迫使我们更有创造性地思考如何研究神经退行性变的机制。人类诱导多能干细胞（iPSC）的出现使我们能够从散发性AD患者以及具有确定家族突变的患者中创建疾病模型。此外，我们可以使用尖端的基因组编辑技术，如Crispr/Cas系统，将疾病相关突变引入健康的人类多能细胞的基因组中。在过去的五年里，神经科学家在从人类来源的多能细胞中创造不同的脑细胞类型，如神经元，星形胶质细胞和少突胶质细胞方面取得了令人难以置信的进展。然而，缺乏的是神经炎症的人类细胞模型，这是所有神经退行性疾病（包括AD）的关键组成部分。在本申请中，我们描述了从患者来源的多能细胞和对照多能细胞产生人类小胶质细胞，大脑的“免疫”细胞。我们提出了一套全面的实验，将确定已知和新的AD相关基因在这些细胞中发挥的作用，使用尖端的基因组编辑技术结合高通量功能测定，转录组学分析和高分辨率蛋白质组学。

DESCRIPTION (provided by applicant): The looming specter of 13.8 million Americans with Alzheimer's disease (AD) by the year 2050 motivates us to expand our biomedical research paradigm outside of the typical "cell line to rodent to human trials" model. The disappointing performance of all AD drugs that have come to Phase III clinical trials to date also forces us to think more creatively about how to study the mechanisms that underlie neurodegeneration. The advent of human induced pluripotent stem cells (iPSCs) allows us to create disease models from patients with sporadic AD as well as from those with defined familial mutations. In addition, we can use cutting-edge genome editing techniques, such as the Crispr/Cas system, to introduce disease-associated mutations into the genome of otherwise healthy human derived pluripotent cells. In the past five years, neuroscientists have made incredible advances in the creation of different brain cell types, such as neurons, astrocytes, and oligodendrocytes, from human-derived pluripotent cells. What is lacking, however, is a human cellular model of neuroinflammation, a critical component of all neurodegenerative disorders, including AD. In the current application, we describe the creation of human microglia, the brain's "immune" cell, from patient-derived and control pluripotent cells. We propose a comprehensive set of experiments that will determine the role that known and novel AD-associated genes play in these cells using cutting-edge genome editing techniques combined with high-throughput functional assays, transcriptomic profiling, and high-resolution proteomics.