Probing Neuropsychiatric Diseases Using Targeted Epigenome and Genome Engineering

摘要 重度抑郁症（MDD）是一种毁灭性的精神疾病，由遗传、 表观遗传和环境影响。尽管经过几十年的研究，我们诊断和治疗的能力 MDD仍然有限，并且大部分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起着重要作用，特别是因为 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开发和电路特异性表观遗传学分析 重塑以逆向工程重性抑郁症的表观遗传机制， 发现新的药物靶点，以开发全新的抗抑郁药。使用动物模型 我们将确定大脑中功能受损的细胞的特定回路， 疾病状态，并确定贡献表观遗传机制。为了使拟议的研究，我们 将开发一个创新的技术平台，以实现靶向基因组和表观基因组修饰 并将其应用于系统地识别特定电路组件中的表观遗传机制， 萧条我们还将探讨已确定的表观遗传机制，以开发新的类别， 抗抑郁药除了我们的基因组和表观基因组工程的核心技术外，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。成功实现我们的愿景将产生四个 广泛影响： I.开创了一种新的药物靶点发现方法，对广泛的 发展和慢性疾病。 二.为大规模靶向基因组工程开发强大的技术平台， 在动物模型中完全重现人类疾病基因型。我们将使精确 将疾病相关基因突变的组合引入单一动物模型。 三.开发靶向表观基因组修饰技术，以实现因果关系的直接功能测试。 特定表观遗传修饰与疾病病理生理学之间的联系。 四.确定治疗抑郁症的新疗法。

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.