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.

项目摘要： 由于缺乏效果，许多影响中枢神经系统（CNS）的遗传疾病仍然无法治疗 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，但是其成功的实施将需要安全的方法来安全 并有效地在整个中枢神经系统中传递基因和基因编辑机械。 AAV是最先进的 体内基因转移的车辆是因为它们可以提供安全且持久的体内基因表达。 AAVS 是唯一已通过调节批准直接给人类行政给予人类的唯一基因治疗载体 美国和欧洲的机构。此外，在2017年，AAV成为第一辆用作早期的一部分 阶段临床试验评估体内基因编辑的安全性。尽管它们令人印象深刻 安全记录，自然发生的AAVS测试迄今为止缺乏大多数基因传递所需的效率 器官系统，包括CNS。为了满足对中枢神经系统基因输送更好的车辆的需求，我们最近 使用的定向进化和一种新的细胞类型在体内选择方法中，以设计几种新颖的AAV， 最值得注意的是AAV-PHP.B和AAV-PHP.EB，这是第一次使其无创的 将基因转移到整个成年小鼠中枢神经系统中的大多数神经元和星形胶质细胞中。在这里，我们的目标是建造 在这种选择方法成功之后，通过工程AAVS来实现整个过程中有效的基因转移 多种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将在几个 物种具有在神经元中提供CNS范围范围范围的转基因表达和靶向基因组编辑的能力，并且 改进的AAV变体将与科学界共享。成功完成该项目，该项目 涉及将新的AAV与下一代基因编辑技术配对，将为评估提供支持 中枢神经系统基因编辑在人体试验中的安全性。