Directed Evolution of Novel AAV Capsis for Global CNS Delivery in Rodents and Primates

标题：新型AAV衣壳在啮齿动物和哺乳动物中用于全球CNS递送的定向进化 灵长 摘要 许多基于单基因的神经系统疾病为基因治疗提供了有吸引力的靶点，但 即使啮齿类动物的概念验证研究前景看好， 在整个中枢神经系统中有效的转基因递送和表达后 （CNS）。定向进化是开发新型腺相关病毒载体的有效方法， 病毒（AAV）载体衣壳，其表现出与天然存在的血清型不同的性质。 然而，到目前为止，大多数新型衣壳都是在啮齿动物或体外模型中获得的 其特性可能会也可能不会转化到其他物种，特别是灵长类动物身上。这 一项利用AAV衣壳DNA改组定向进化来开发基因递送的提案 用于许多人类CNS疾病应用的载体。为此，并行选择和 将在小鼠和非人灵长类动物（NHP）中进行回收的克隆表征， 结合NHP中CNS基因转移和AAV载体设计专家的专业知识。 实验计划应独立地产生能够全局表达的上级AAV衣壳。 CNS递送，在小鼠和NHP之间具有交叉相容性。此外，我们还设计了 我们的实验方法是产生对神经元表现出选择性向性的载体， 星形胶质细胞和/或少突胶质细胞，这将是研究和 治疗应用。如果成功，产生的AAV衣壳试剂应该产生新的 研究工具，扩大基因治疗的应用，以更多的中枢神经系统疾病，并促进 将现有的中枢神经系统基因治疗方法转化为人类。

TITLE: Directed Evolution of Novel AAV Capsids for Global CNS Delivery in Rodents and Primates ABSTRACT Many monogenic based neurological disorders present attractive targets for gene therapy, but even with promising proof-of-concept rodent studies, successful clinical translation depends upon efficient transgene delivery and expression across the entire central nervous system (CNS). Directed evolution is a powerful and proven method to develop novel adeno-associated virus (AAV) vector capsids that exhibit properties distinct from naturally occurring serotypes. However, to date, the majority of novel capsids have been derived in rodents or in vitro models whose properties may or may not translate to other species, in particular primates. This proposal will utilize AAV capsid DNA shuffling directed evolution to develop gene delivery vectors for a number of human CNS disease applications. To do this, parallel selections and recovered clone characterization will be carried out in mice and non-human primates (NHPs), combining the expertise of experts on both CNS gene transfer in NHPs and AAV vector design. The experimental plan should independently generate superior AAV capsids capable of global CNS delivery, with cross-compatibility between mice and NHPs. Moreover, we have designed our experimental approach to generate vectors that exhibit selective tropism for neurons, astrocytes, and/or oligodendrocytes, which would be invaluable reagents for research and therapeutic applications. If successful, the AAV capsid reagents generated should create new research tools, broaden the application of gene therapy to more CNS disorders, and facilitate the translation of existing CNS gene therapy approaches to humans.