Probing Neuropsychiatric Diseases Using Targeted Epigenome and Genome Engineering

摘要 严重抑郁障碍(MDD)是一种毁灭性的精神疾病，由遗传、 表观遗传和环境影响。尽管经过了几十年的调查，我们诊断和治疗的能力 MDD仍然有限，很大一部分MDD患者对可用的治疗方案没有反应。 表观遗传基因组变化可能在抑郁症的病理生理学中发挥重要作用，特别是在 环境刺激和经验是MDD发展的重要因素。我们建议 基因组甲基化和染色质的偶联技术发展和电路特异性表观遗传分析 重塑以逆转重大抑郁障碍背后的表观遗传机制，并 发现新的药物靶点，以开发从根本上新型的抗抑郁药物。使用动物模型 对于抑郁症，我们将识别大脑中特定的细胞回路，其功能在 疾病状态，并确定起作用的表观遗传机制。为了支持拟议的研究，我们 将开发一个创新的技术平台，使目标基因组和表观基因组修改成为可能 并将其应用于系统地识别特定电路组件的表观遗传机制 抑郁症。我们还将探索已确定的表观遗传机制，以发展新的类别 抗抑郁药。除了我们的基因组和表观基因组工程核心技术外，我们还将整合 全面的技术专长，涵盖电生理学、成像、行为分析、 计算生物学、合成生物学、高通量基因组和表观基因组分析以及高通量 药物筛选和分析开发。我们的愿景的成功完成将产生四个 广泛的影响： I.开拓者一种新的药物靶标发现方法，对广泛的 发育性和慢性病。 为大规模靶向基因组工程开发强大的技术平台，以实现更多 动物模型中人类疾病基因分型的完整概括。我们将使精确度 将与疾病相关的基因突变组合引入单一动物模型。 开发一种有针对性的表观基因组修饰技术，以实现对因果关系的直接功能测试 特定的表观遗传修饰和疾病病理生理学之间的联系。 从根本上找出治疗重度抑郁症的新疗法。

Abstract Major depressive disorder (MDD) is a devastating mental illness arising from a combination of genetic, epigenetic, and environmental influences. Despite decades of investigation, our ability to diagnose and treat MDD remains limited, and a large fraction of MDD patients fail to respond to available treatment options. Epigenetic genome changes likely play a significant role in the pathophysiology of depression, especially since environmental stimuli and experience are important contributors to the development of MDD. We propose to couple technology development and circuit-specific epigenetic analysis of genome methylation and chromatin remodeling to reverse engineer the epigenetic mechanisms underlying major depressive disorder, and to discover novel drug targets for developing fundamentally new classes of antidepressants. Using animal models of depression we will identify specific circuits of cells in the brain whose functions are compromised in the disease state and determine the contributing epigenetic mechanisms. To enable the proposed research, we will develop an innovative platform of technologies to enable targeted genome and epigenome modifications and apply it to systematically identify epigenetic mechanisms in specific circuit components underlying depression. We will also explore the identified epigenetic mechanisms for developing new classes of antidepressants. In addition to our core technologies for genome and epigenome engineering, we will integrate a comprehensive range of technical expertise spanning electrophysiology, imaging, behavioral analysis, computational biology, synthetic biology, high-throughput genome and epigenome analysis, and highthroughput drug screening and assay development. The successful completion of our vision will yield four broad impacts: I. Pioneer a new approach for drug target discovery that has implications for a broad range of developmental and chronic illnesses. II. Develop a robust technology platform for large-scale targeted genomic engineering to enable more complete recapitulation of human disease genotypes in animal models. We will enable precise introduction of combinations of disease-associated genetic mutations into a single animal model. III. Develop a technology for targeted epigenome modification to enable direct functional testing of causal links between specific epigenetic modifications and disease pathophysiology. IV. Identify fundamentally new classes of therapeutics for major depression.