Ionic liquid-assisted drug delivery to brain reservoirs for treatment of neuroHIV

项目总结/摘要 尽管抗逆转录病毒疗法（ART）取得了成功，但它们无法根除艾滋病毒的水库内， 身体，特别是中枢神经系统（CNS）。此外，阿片类药物上调外排转运蛋白 进一步促进CNS中ART的亚治疗浓度。我们使用生物相容性离子液体 (ILs)，由不对称阳离子和阴离子组成的熔融盐，可以“调节”纳米颗粒的亲和力， 不同的细胞类型使用这种策略，我们已经开发了对红细胞具有平衡亲和力的IL， 小胶质细胞促进纳米颗粒在红细胞上“搭便车”，将它们运送到大脑，细胞- 一旦递送到中央室，选择性靶向小胶质细胞。大鼠的初步数据表明， ~48%的注射纳米粒子在6小时内在大脑中积累，这是一个巨大的进步， 纳米颗粒递送策略。初步分析表明，超过90%的CNS纳米颗粒是 与小胶质细胞有关。我们进一步证明了将ART（阿巴卡韦）装载到纳米颗粒中的能力。 当给予HIV感染的人外周血单核细胞时， （PBMC）。我们假设，我们可以进一步改善我们的IL制剂的可调概况，以优化货物 递送到大脑并靶向其他细胞类型（包括星形胶质细胞）。我们预计这批货物 尽管阿片类药物促进了CNS细胞粘附/转运蛋白的变化， 暴露/依赖。为此，我们将（目标1）产生至少5个新的离子液体，除了我们目前的领导， 具有不同的细胞类型亲和力。我们将确认IL赋予猿和人类血液的偏好 作为潜在的货物运输工具。(Aim 2）我们将评估安全性（亚急性，急性，亚慢性， 生殖、致突变）和生物分布，最多5种新型IL在阿片类药物初治或阿片类药物依赖大鼠中的分布。 IL将装载有预制的具有eGFP报告基因的scramble Cas9载体，以确认递送细胞的能力。 CRISPR-Cas9构建体用于潜在的HIV治愈策略。具有CNS有利生物分布的安全IL将是 加载cART（阿巴卡韦，度鲁特韦和拉米夫定），并评估其在SIV感染或HIV-1感染患者中的疗效。 感染的猿猴或人PBMC，或人小胶质细胞。所有细胞都是阿片样物质未用过的或阿片样物质未用过的。 暴露了将优先考虑对小胶质细胞和星形胶质细胞具有选择性的IL。(Aim 3）IL导联（基于CNS 分布和小胶质细胞/星形胶质细胞选择性）的体内安全性、生物分布和急性毒性。 在SIV恒河猴模型中的有效性。在猕猴中，IL辅助的纳米颗粒将装载纳米金， 除cART外，通过X射线确认生物分布的时间进程。SIV感染将通过 脑脊液和血液样本。将对血浆进行全血细胞计数、化学和细胞因子分析 和/或CSF。将通过LC/MS确认ART分布。将对器官进行大体组织病理学检查 另外评估CNS组织的小胶质细胞增生、星形胶质细胞增生和亚致死性神经元损伤。白介素- 辅助纳米颗粒可以实现安全、细胞特异性、中枢神经系统药物/货物递送的目标。

PROJECT SUMMARY/ABSTRACT Despite the success of antiretroviral therapeutics (ARTs), they cannot eradicate HIV from reservoirs within the body, particularly those in the central nervous system (CNS). Moreover, opioids upregulate efflux transporters further promoting subtherapeutic concentrations of ART in the CNS. We have used biocompatible ionic liquids (ILs), molten salts comprised of asymmetric cations and anions, that can `tune' the affinity of nanoparticles to different cell types. Using this strategy, we have developed an IL with a balanced affinity for erythrocytes and microglia which promotes nanoparticle `hitchhiking' on erythrocytes to deliver them to the brain, and cells- selective targeting of microglia once delivered to the central compartment. Preliminary data in rats demonstrate ~48% of injected nanoparticles accumulating in the brain within 6 hours, a vast improvement over current nanoparticle delivery strategies. Preliminary analyses indicated that over 90% of CNS nanoparticles were associated with microglia. We have further demonstrated the capacity to load ART (abacavir) into nanoparticles which retained antiviremic efficacy when administered to HIV-infected human peripheral blood mononuclear cells (PBMCs). We hypothesize that we can further improve the tunable profile of our IL formulation to optimize cargo delivery to the brain and target additional cell types (including astrocytes). We anticipate that this cargo delivery strategy will be safe and efficacious in spite of CNS cell adhesion/transporter changes promoted by opioid exposure/dependence. To this end, we will (Aim 1) generate at least 5 novel ILs, in addition to our current lead, with varied cell-type affinity. We will confirm the preference that ILs confer to simian and human blood components as potential cargo carriers. (Aim 2) We will assess the safety (subacute, acute, subchronic, reproductive, mutagenic) and biodistribution of up to 5 novel ILs in rats that are opioid-naïve or opioid-dependent. ILs will be loaded with a premade scramble Cas9 vector with an eGFP reporter to confirm the capacity to deliver CRISPR-Cas9 constructs for potential HIV cure strategies. Safe ILs with a CNS-favorable biodistribution will be loaded with cART (abacavir, dolutegravir, and lamivudine) and assessed for efficacy in SIV-infected or HIV- infected simian or human PBMCs, respectively, or human microglia. All cells will be opioid-naïve or opioid- exposed. ILs with selectivity for microglia and astrocytes will be prioritized. (Aim 3) IL leads (based on CNS distribution and microglial/astrocytic selectivity) will be assessed for in vivo safety, biodistribution, and acute efficacy in a rhesus macaque model of SIV. In macaques, IL-assisted nanoparticles will be loaded with nanogold, in addition to cART, to confirm the time-course of biodistribution via X-ray. SIV infection will be monitored via CSF and blood draws. Complete blood count, chemistry, and cytokine profiling will be conducted on plasma and/or CSF. ART distribution will be confirmed via LC/MS. Gross histopathology will be conducted on organs and CNS tissues will be additionally assessed for microgliosis, astrogliosis, and sublethal neuronal damage. IL- assisted nanoparticles may realize the goal of achieving safe, cell-specific, CNS drug/cargo delivery.