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

专为整个中枢神经系统进行高效、非侵入性跨物种基因编辑而设计的新型 AAV

• 批准号: 10490394
• 负责人: Benjamin E Deverman
• 金额: $ 135.97万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10490394
• 关键词: Address; Adult; Affect; Animal Testing; Animals; Astrocytes; Biological Products; Blood - brain barrier anatomy; Brain; Callithrix; Capsid; Cells; Central Nervous System Diseases; Characteristics; Clinical; Clinical Trials; Clinical Trials Design; Collaborations; Communities; Dependovirus; Directed Molecular Evolution; Disease; Engineering; Ensure; Enterobacteria phage P1 Cre recombinase; Europe; Evaluation; Future; Gene Delivery; Gene Expression; Gene Transduction Agent; Gene Transfer; Genes; Genetic; Genetic Diseases; Genetic Models; Human; Injections; Intravenous; Knock-in; Lead; Libraries; Macaca mulatta; Mental disorders; Methods; Methyl-CpG-Binding Protein 2; Mucopolysaccharidosis II; Mus; Nature; Neuraxis; Neuroglia; Neurons; Pharmaceutical Preparations; Phase; Population; Production; Rattus; Recovery; Reporter; Rhesus; Rodent; Safety; Somatic Cell; System; Technology; Testing; Therapeutic; Time; Toxic effect; United States National Institutes of Health; Variant; Virus; base; body system; cell type; early phase clinical trial; gene delivery system; gene therapy; genome editing; genome-wide; improved; in vivo; insertion/deletion mutation; interest; intravenous administration; lead candidate; nervous system disorder; next generation; nonhuman primate; novel; pre-clinical; programs; scale up; small molecule; somatic cell gene editing; success; tool; transduction efficiency; transgene expression; vector; vector genome

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.

项目概要： 由于缺乏有效的治疗方法，许多影响中枢神经系统（CNS）的遗传性疾病仍然无法治疗。 小分子药物或生物制剂。利用体细胞基因针对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 并有效地将基因和基因编辑机器传递到整个中枢神经系统。 AAV 是最先进的 体内基因转移的载体，因为它们可以提供安全且持久的体内基因表达。腺相关病毒 是唯一已被监管部门批准直接用于人类的基因治疗载体 美国和欧洲的机构。此外，2017 年，AAV 成为第一款用作早期 阶段临床试验评估体内基因编辑的安全性。尽管它们的临床前和临床效果令人印象深刻 安全记录显示，迄今为止测试的天然 AAV 缺乏大多数基因传递所需的效率 器官系统，包括中枢神经系统。为了满足对中枢神经系统基因传递更好的载体的需求，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设计几种新型 AAV， 最值得注意的是 AAV-PHP.B 和 AAV-PHP.eB，它们首次使非侵入性 将基因转移到成年小鼠中枢神经系统的大多数神经元和星形胶质细胞。在这里，我们的目标是打造 这种选择方法成功后，通过工程 AAV 实现了高效的基因转移 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 AAV 将在多个方面进行评估 物种能够提供中枢神经系统范围内的转基因表达和神经元中的靶向基因组编辑，以及 改进的 AAV 变种将与科学界共享。该项目的顺利完成， 涉及将新的 AAV 与下一代基因编辑技术配对，将为评估提供支持 中枢神经系统基因编辑在人体试验中的安全性。