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

标题：用于全球中枢神经系统递送的新型AAV衣壳在啮齿动物和 灵长类 摘要 许多基于单基因的神经疾病为基因治疗提供了诱人的靶点，但 即使有很有前景的概念验证啮齿动物研究，成功的临床翻译也取决于 在整个中枢神经系统中高效地传递和表达转基因 (CNS)。定向进化是发展新腺相关基因的一种有效且已被证实的方法 病毒(AAV)载体衣壳显示出与自然发生的血清型不同的特性。 然而，到目前为止，大多数新型衣壳都是从啮齿动物或体外模型中衍生出来的。 其特性可能会也可能不会转化为其他物种，特别是灵长类动物。这 提案将利用AAV衣壳DNA改组定向进化来发展基因传递 用于多种人类中枢神经系统疾病的病媒。要做到这一点，并行选择和 恢复的克隆特征将在小鼠和非人灵长类动物(NHP)中进行， 结合NHP中CNS基因转移和AAV载体设计方面的专家的专业知识。 实验计划应独立生成能够在全球范围内 中枢神经系统递送，具有小鼠和NHP之间的交叉相容性。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取向的载体， 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衣壳试剂应该会产生新的 研究工具，拓宽基因治疗的应用范围，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.