Next-generation extracellular vesicle biologics to target central nervous system and peripheral reservoirs of HIV

摘要 在联合抗逆转录病毒治疗 (cART) 时代，与 HIV-1 相关的死亡率和发病率 感染已减少。然而，与艾滋病相关的多种病症以及严重的 非艾滋病事件 (SNAE) 继续困扰着艾滋病毒感染者 (PLWH)。长期改善 需要采取一些策略，从更容易的给药到更好的抑制，再到功能性治愈。 功能性治愈的障碍包括输送活化剂和其他生物活性分子的能力有限 进入 HIV 的组织储存库，特别是中枢神经系统 (CNS) 中可能的储存库。 细胞外囊泡 (EV) 是双膜结合的，代表一种新颖的递送模式 由所有已知细胞类型释放的颗粒，通过穿梭参与细胞间通讯 将亲本细胞的成分（例如蛋白质和RNA）转移到靶细胞。 EV 会导致疾病 发病机制并被积极研究，特别是在癌症中，作为生物标志物、疾病的参与者 过程和潜在的治疗方法。重要的是，电动汽车已被证明可以跨生物 屏障，甚至是血脑屏障，并且可以很容易地传递到大脑。因此，电动汽车提供了 向组织储存库提供艾滋病毒控制或重新激活/治愈成分的绝佳机会， 具有细胞特异性的潜力。 为此，我们组建了一支拥有两大独特资产的多学科团队。第一个是 新型小型 EV 转录激活剂 (sEVTA) 工具已通过体外和体内试验 测试。在该系统中，逆转录病毒反式激活蛋白被专门包装到 EV 中，从而可以 通过示踪剂和表面肽进一步功能化以进行细胞靶向。第二个是我们的井 建立了SIV/猕猴模型，已成功用于研究逆转录病毒潜伏期， 反弹和逆转录病毒相关的中枢神经系统疾病。我们将仔细进行非人类灵长类动物给药 和分布研究，然后优化 sEVTA 的静脉内和鼻内输送。效果 将使用循环和外周组织的创新工具评估 sEVTA 对病毒反弹的影响 水库（目标 1），然后是中枢神经系统中的水库（目标 2）。在目标 3 中，我们将调查 这些方法的潜在毒性，特别是对中枢神经系统的潜在毒性，以及 探索更多的细胞靶向方法。 这些研究的目标是利用 sEVTA 重新激活中枢神经系统和大脑中潜伏的逆转录病毒。 外围。然而，该项目还将提供有关电动汽车交付的急需信息， 灵长类动物中的分布和功效可用于开发多种疗法 用于艾滋病毒感染和疾病。

Abstract In the era of combined antiretroviral therapy (cART), mortality and morbidity associated with HIV-1 infection has been reduced. Nevertheless, a wide range of AIDS-related conditions as well as serious non-AIDS events (SNAEs) continue to afflict people living with HIV (PLWH). Improved long-term strategies are needed, ranging from easier administration to better suppression to functional cure. Barriers to functional cure include a limited ability to deliver activators and other bioactive molecules into tissue reservoirs of HIV, particularly the possible reservoirs in the central nervous system (CNS). Representing a novel mode of delivery, extracellular vesicles (EVs) are double membrane-bound particles, released by all known cell types, that engage in intercellular communication by shuttling components of the parent cell (such as proteins and RNAs) to target cells. EVs contribute to disease pathogenesis and are actively investigated, especially in cancers, as biomarkers, actors in disease processes, and potential therapeutics. Importantly, EVs have been shown to cross biological barriers, even the blood-brain barrier, and can be easily delivered to the brain. EVs thus provide an exceptional opportunity to deliver components of HIV control or reactivation/cure to tissue reservoirs, with potential for cell-specificity. To this end, we have assembled a multidisciplinary team with two major, unique assets. The first is a novel small EV-transcriptional activator (sEVTA) tool that has already passed in vitro and in vivo tests. In this system, retroviral transactivator proteins are specifically packaged into EVs, which can be further functionalized with tracers and surface peptides for cell targeting. The second is our well established SIV/macaque model, which has been used successfully to study retroviral latency, rebound, and retrovirus-associated CNS disease. We will conduct careful nonhuman primate dosing and distribution studies followed by optimized intravenous and intranasal delivery of sEVTAs. Effects of sEVTAs on viral rebound will be assessed with innovative tools in circulating and peripheral tissue reservoirs (Aim 1), followed by reservoirs in the CNS (Aim 2). In Aim 3, we will investigate the potential toxicity of these approaches, with particular focus on the central nervous system, and explore more cell-targeted approaches. The goal of these studies is to use sEVTAs to reactivate latent retroviruses in the CNS and the periphery. However, the project will also provide much-needed information about EV delivery, distribution, and efficacy in primates that can be harnessed in development of a variety of therapies for HIV infection and disease.