Evolving Novel AAV Vectors for Gene Therapy to Cure HIV

项目总结 鉴于目前感染艾滋病毒的人数是历史上最多的，遏制艾滋病毒流行仍是当务之急。 联合抗逆转录病毒疗法(ART)可以限制病毒复制，但不能治愈。因此，迫切需要 通过清除病毒库来设计一种功能性治愈方法。蒂莫西·布朗，也就是柏林病人和亚当 卡斯蒂略，也就是伦敦的患者，在与白血病相关的、MHC匹配的异基因移植后治愈了艾滋病毒 CCR5缺陷供者的造血干细胞移植(HSCT)。而CCR5缺陷的免疫系统 系统可以明显地产生功能性的HIV治愈，异基因干细胞移植不能扩展到 需要总体人口和替代方法。我们已经证明了CCR5特定的 Leronlimab抗体可以通过占据所有可用的CCR5来在药理学上模拟CCR5缺陷的供体 分子。为了将Leronlimab作为一种基因治疗选择，需要新的给药方式。这里, 我们建议利用我们的新的定向进化技术来生成特定于T和B的AAV载体 细胞。这些新型AAV载体将促进Leronlimab在体内的表达，但更重要的是 将支持未来使用其他抗艾滋病毒方法，包括CRISPR-Cas9、嵌合抗原受体、 并通过将这些治疗药物输送到相关的免疫细胞类型来广泛中和抗体。具体而言 目标1，我们将产生并鉴定携带AAV衣壳的靶向T和B细胞的衣壳 猕猴和人类。在目标2中，我们将通过以下方式演示这些新AAV的概念验证效用 Leronlimab用于感染SHV、ART抑制的猕猴，以确定是否可以通过 这种方法。这项工作将扩大我们对HIV治愈机制的了解，因为它展示了 基于抗体的长期竞争抑制CCR5并建立一套新的AAV载体以支持体内 提供抗艾滋病毒治疗药物。

PROJECT SUMMARY With the most people ever in history currently living with HIV, stopping the HIV epidemic remains imperative. Combination antiretroviral therapy (ART) limits viral replication, but is not curative. Thus, there is an urgent need to design a functional cure via elimination of the viral reservoir. Timothy Brown, aka the Berlin Patient, and Adam Castillejo, aka the London patient, were cured of HIV following leukemia-related, MHC-matched, allogeneic hematopoietic stem cell transplantation (HSCT) from a CCR5-deficient donor. While a CCR5-deficient immune system can demonstrably yield a functional HIV cure, allogeneic stem cell transplantation is not scalable to the general population and alternate approaches are needed. We have demonstrated that the CCR5-specific antibody Leronlimab can pharmacologically mimic a CCR5 deficient donor by occupying all available CCR5 molecules. In order to deliver Leronlimab as a gene therapy option, new delivery modalities are needed. Here, we are proposing to utilize our novel directed evolution technique to generate AAV vectors specific for T and B cells. These novel AAV vectors will facilitate in vivo delivery of Leronlimab expression here, but more importantly will support the future use of other anti-HIV approaches including CRISPR-Cas9, chimeric antigen receptors, and broadly neutralizing antibodies by delivering these therapeutics to the relevant immune cell type. In specific aim 1, we will generate and characterize AAV bearing capsids that target T and B cells specifically across both macaques and humans. In aim 2, we will demonstrate proof-of-concept utility of these new AAVs by delivering Leronlimab to SHIV-infected, ART suppressed macaques to determine if a functional cure can be achieved with this approach. This work would expand our knowledge of the mechanism of HIV cure by showing the utility of long-term antibody-based competitive CCR5 inhibition and establish a new set of AAV vectors to support in vivo delivery of anti-HIV therapeutics.