An AAV-mediated functional cure and its impact on the reservoir

计划摘要 在初步数据中，我们展示了两项研究的结果，这两项研究表明，持续的无艺术状态 在非人类灵长类动物中，病毒学缓解是可能的，这是一种“功能治愈”。首先，Desrosiers实验室 已经表明，两种广谱中和抗体(BNAbs)的长期表达可以抑制 三只恒河猴中未经治疗的SHV-AD8感染，在一例中，检测不到(15拷贝/毫升) 两年多了。其次，Farzan实验室已经证明，长期表达的抗体样 进入抑制剂eCD4-Ig也能有效地抑制六个SHV-AD8感染的恒河猴中的五个的病毒复制 猕猴停止抗逆转录病毒治疗后一年多(ART)。我们建议在此基础上 这些初步结果使这些功能治疗更安全、更一致和更强大。在 过程中，我们将建立一个有用的平台来评估延迟反转代理(LRA)，即所谓的 ‘踢’，通过提供一个强有力的‘杀戮’始终存在的环境，并开发一种方法来阻止 转基因表达，从而实现反弹时间测量。我们将确定如何最好地 限制AAV表达的bNAbs产生的抗药物抗体(ADA)。我们将确定如何最好地 使用ART或长效整合酶抑制剂卡替格列韦来建立AAV介导的功能性治疗。 最后，我们将确定持续和有效的杀戮本身是否可以改变潜伏的衰减率 被感染的细胞。为此，我们组建了一支在艾滋病毒和SIV研究方面具有丰富经验的团队，多年来 共同工作的经验，以及长期致力于理解和改进基于AAV的 功能性治疗。这些工作是通过四个项目和三个核心完成的。项目1将建立 使用AAV表达的eCD4-Ig对感染SIV-AD8和SIVmac239的猕猴进行强大的功能治疗 并评估持续的eCD4-Ig对病毒库的影响。项目2将开发和测试多个 消除经常与AAV表达的bNAbs一起出现的抗药物抗体的方法。项目 3将评估三联疗法和长效卡替格雷对eCD4-Ig建立的影响。 介导性功能性治疗，并确定卡替格列韦与eCD4-Ig的最佳结合方式。项目 4将开发一种永久的AAV转基因开关，促进反弹时间研究 持续表达bNAb，增加AAV介导的功能性治疗的安全性。这些 项目围绕着支持一系列非人类生物的统一检测实验流水线进行组织 灵长类研究，由核心B建立和实施。核心A将管理监管和后勤 该计划的各个方面，核心C将为这些研究提供生产高质量的AAV颗粒，以及 提高其这样做的能力。总体而言，这些努力将发展和改进一种可行的方法 对人类的有效治疗，并为彻底根除病毒提供有用的工具和洞察力。

PROGRAM SUMMARY In preliminary data we show the results of two studies demonstrating that a sustained state of ART-free virologic remission, a ‘functional cure’, is possible in non-human primates. First, the Desrosiers laboratory has shown that long-term expression of two broadly neutralizing antibodies (bNAbs) can suppress an untreated SHIV-AD8 infection in three rhesus macaques, in one case, to undetectable (<15 copies/ml) for over two years. Second, the Farzan laboratory has shown that long-term expression of the antibody-like entry inhibitor eCD4-Ig can also efficiently suppress viral replication in five of six SHIV-AD8-infected rhesus macaques for more than a year after cessation of anti-retroviral therapy (ART). We propose to build on these initial results by making these functional cures safer, more consistent, and more robust. In the process, we will establish a useful platform for evaluating latency-reversing agents (LRAs), a so-called ‘kick’, by providing an environment in which a potent ‘kill’ is always present, and develop a way to halt transgene expression, thereby enabling time-to-rebound measurements. We will determine how best to limit anti-drug antibodies (ADA) that emerge with AAV-expressed bNAbs. We will determine how to best use ART or the long-lasting integrase inhibitor cabotegravir to establish AAV-mediated functional cures. Finally, we will determine whether a sustained and potent kill can by itself change the decay rate of latently infected cells. To do so, we have assembled a team with deep experience in HIV and SIV studies, years of experience working together, and a long-term commitment to understanding and improving AAV-based functional cures. These effort are accomplished with four projects and three cores. Project 1 will establish robust functional cures in SHIV-AD8- and SIVmac239-infected macaques using AAV-expressed eCD4-Ig and evaluate the impact of sustained eCD4-Ig on the viral reservoir. Project 2 will develop and test multiple approaches for eliminating anti-drug antibodies that frequently emerge with AAV-expressed bNAbs. Project 3 will assess the impact of triple therapy and long-acting cabotegravir on the establishment of eCD4-Ig- mediated functional cures, and determine how cabotegravir might best be combined with eCD4-Ig. Project 4 will develop a permanent off- and on-switches for AAV transgenes, facilitating time-of-rebound studies after sustained bNAb expression and increasing the safety of AAV-mediated functional cures. These projects are organized around a uniform experimental pipeline of assays supporting a series of non-human primate studies, established and implemented by Core B. Core A will manage regulatory and logistical aspects of the Program, and Core C will provide produce high-quality AAV particles for these studies, and improve its capacity for doing so. Collectively these efforts will develop and improve a viable approach to functional cures in humans, and provide tools and insight useful for complete eradication of the virus.