Evolving Novel AAV Vectors for Gene Therapy to Cure HIV

项目摘要 由于目前艾滋病毒感染者人数达到历史上最多的水平，阻止艾滋病毒的流行仍然是当务之急。 联合抗逆转录病毒疗法（ART）限制病毒复制，但不能治愈。因此，迫切需要 通过消除病毒库来设计功能性治疗方法。蒂莫西·布朗又名柏林病人和亚当 卡斯蒂列霍，又名伦敦病人，在白血病相关的，MHC匹配的，同种异体移植后， 造血干细胞移植（HSCT）从CCR 5缺陷的供体。而缺乏CCR 5的免疫系统 系统可以证明产生功能性HIV治愈，同种异体干细胞移植不能扩展到 需要一般人群和替代方法。我们已经证明，CCR 5特异性 抗体Leronlimab可以通过占据所有可用的CCR 5， 分子。为了将Leronlimab作为一种基因治疗选择，需要新的递送方式。在这里， 我们建议利用我们新的定向进化技术来产生T和B特异性的AAV载体 细胞这些新的AAV载体将促进Leronlimab表达的体内递送，但更重要的是， 将支持未来使用其他抗HIV方法，包括CRISPR-Cas9，嵌合抗原受体， 以及通过将这些治疗剂递送至相关的免疫细胞类型来广泛中和抗体。在特定 目的1，我们将产生和表征携带AAV衣壳，其特异性靶向T和B细胞， 猕猴和人类在目标2中，我们将通过交付这些新的无人驾驶飞机来证明概念实用性。 Leronlimab对SHIV感染的ART抑制猕猴的作用，以确定是否可以通过以下方法实现功能性治愈： 这种方法。这项工作将扩大我们对艾滋病毒治愈机制的认识， 长期基于抗体的竞争性CCR 5抑制，并建立一组新的AAV载体，以支持体内 递送抗HIV治疗剂。

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.