Novel AAVs Engineered for Efficient and Noninvasive Cross-Species Gene Editing Throughout the Central Nervous System

项目概要： 许多影响中枢神经系统（CNS）的遗传性疾病由于缺乏有效的治疗方法而无法治疗。 小分子药物或生物制剂。用体细胞基因靶向这些疾病的遗传基础 因此，编辑将特别有影响力，但其成功实施将需要安全的方法， 并在整个CNS中有效地传递基因和基因编辑机制。自动驾驶飞机是最先进的 载体，因为它们可以提供安全和持久的体内基因表达。自动增值 是唯一被监管机构批准用于直接给药于人类的基因治疗载体 美国和欧洲的代理商。此外，在2017年，无人驾驶飞机成为第一辆作为早期交通工具的一部分使用的车辆。 阶段临床试验，以评估体内基因编辑的安全性。尽管他们令人印象深刻的临床前和临床 安全记录，迄今为止测试的天然存在的AAV缺乏基因递送所需的效率， 器官系统，包括CNS。为了解决对更好的CNS基因递送载体的需求，我们最近 使用定向进化和新的细胞类型特异性体内选择方法来工程化几种新的AAV， 最值得注意的是AAV-PHP.B和AAV-PHP.eB，这是第一次，使非侵入性地 将基因转移到整个成年小鼠CNS的大多数神经元和星形胶质细胞。在这里，我们的目标是 在通过工程化能够在整个细胞中进行有效基因转移的AAV的这种选择方法成功后， 多个物种的中枢神经系统，包括非人类灵长类动物。我们开发的无人驾驶飞机将在几个 物种，因为它们能够在神经元中提供CNS范围的转基因表达和靶向基因组编辑， 改进的AAV变体将与科学界分享。成功完成该项目， 涉及将新的AAV与下一代基因编辑技术配对，将为评估 CNS基因编辑在人体试验中的安全性。

Project Summary: Many genetic diseases that affect the central nervous system (CNS) remain untreatable due to a lack effective small molecule drugs or biologics. Targeting the genetic underpinnings of these diseases with somatic cell gene editing would therefore be particularly impactful, but its successful implementation will require methods to safely and efficiently deliver genes and gene editing machinery throughout the CNS. AAVs are the state-of-the-art vehicles for in vivo gene transfer because they can provide safe and long lasting in vivo gene expression. AAVs are the only gene therapy vectors that have been approved for direct administration to humans by regulatory agencies in both the US and Europe. Moreover, in 2017, AAVs became the first vehicle used as part of an early phase clinical trial to evaluate the safety of in vivo gene editing. Despite their impressive preclinical and clinical safety record, naturally occurring AAVs tested to date lack the efficiency required for gene delivery across most organ systems, including the CNS. To address the need for better vehicles for CNS gene delivery, we recently used directed evolution and a new cell type-specific in vivo selection method to engineer several novel AAVs, most notably AAV-PHP.B and AAV-PHP.eB, that have, for the first time, made it possible to noninvasively transfer genes to the majority of neurons and astrocytes throughout the adult mouse CNS. Here, we aim to build upon the success of this selection approach by engineering AAVs that enable efficient gene transfer throughout the CNS of multiple species, including nonhuman primates. The AAVs we develop will be evaluated in several species for their ability to provide CNS-wide transgene expression and targeted genome editing in neurons, and improved AAV variants will be shared with the scientific community. Successful completion of this project, which involves pairing the new AAVs with next-generation gene editing technologies, will provide support for evaluating the safety of CNS gene editing in human trials.