Dual antiretroviral loaded nanoparticles for long-acting suppressive HIV therapy

用于长效抑制艾滋病毒治疗的双重抗逆转录病毒纳米粒子

• 批准号: 10548482
• 负责人: Josiane Fofana
• 金额: $ 4.68万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-28 至 2025-02-27
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10548482
• 关键词: AIDS prevention; Address; Adherence; Anti-Retroviral Agents; Binding; Biodistribution; Biotin; CD4 Positive T Lymphocytes; Cell Survival; Cell membrane; Cell surface; Cells; Chronic; Coculture Techniques; Data; Dendritic Cells; Development; Disease; Disseminated Malignant Neoplasm; Dose; Dyes; Exposure to; Failure; Flow Cytometry; G(M3) Ganglioside; Goals; HIV; HIV therapy; HIV-1; Image Cytometry; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Inflammatory Response Pathway; Injections; Integrase Inhibitors; Label; Lead; Lectin Receptors; Ligase; Link; Lipid Bilayers; Lipids; Lymphoid Tissue; Measures; Mediating; Membrane; Morbidity - disease rate; Mus; Myelogenous; Myeloid Cells; Oral; Patients; Penetration; Pharmaceutical Preparations; Phenotype; Plasma; Polymers; Prevention; Property; Protein-Protein Interaction Map; Regimen; Reverse Transcriptase Inhibitors; Risk; Role; Secondary to; Sialic Acids; Site; Spatial Distribution; Synapses; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Toxic effect; Viral; Virus; antiretroviral therapy; combinatorial; comorbidity; cytokine; design; extracellular; immune activation; immunoregulation; in vivo; innovation; live cell imaging; macrophage; monocyte; nanoformulation; nanoparticle; nanotherapeutic; non-nucleoside reverse transcriptase inhibitors; nonhuman primate; novel strategies; poly(lactic acid); preservation; prevent; response; targeted delivery; transmission process; uptake; viral rebound; viral transmission; virological synapse

PROJECT SUMMARY/ABSTRACT HIV remains a global threat despite the tremendous progress of combinatorial antiretroviral therapy (cART). Chronic inflammation and immune activation support an increased risk of non-AIDS co-morbidities in cART- treated HIV patients. This condition is attributed to various factors including antiretroviral (ARV) toxicity and viral persistence in secondary lymphoid tissues (SLTs) which has been linked to low ARV penetration in these reservoir sites. Furthermore, uptake of daily antiretrovirals (ARVs) remains a burden resulting in poor adherence that can lead to viral rebound from the SLTS. In this project, we propose to develop a long-acting “stealth” nanoformulation composed of lipid wrapped poly-lactic acid nanoparticles (GM3-PLA-NPs) loaded with a non-nucleoside reverse transcriptase inhibitor, Rilpivirine (RPV) and integrase inhibitor, Cabotegravir (CAB). These GM3-PLA-NPs have innovative targeting and design features that will increase drug entry and retention in SLTs, in addition to modulating inflammatory responses. Indeed, the membrane incorporated monosialo di-hexosylganglioside (GM3) enables specific binding to the sialic acid lectin receptor CD169 on myeloid cells (macrophages and dendritic cells), leading to the formation of non-acidic membrane invaginations which can prevent NP degradation, and lead to long-term establishment of cellular drug depots. These NP collecting compartments (NPCCs) are identical to the previously defined virus containing compartments (VCCs) formed upon capture of HIV-1 by CD169. Since VCCs are also involved in myeloid cell- mediated HIV transmission to CD4+ T cells (trans-infection) through the virological synapse, I hypothesize that NPCCs could potentially promote sustained ARV release to bystander T cells through macrophage – T cell synaptic junctions. Our preliminary data have demonstrated that these GM3-PLA-NPs are successfully preserved in NPCCs of CD169+ macrophages over time which extended anti-viral potency in these cells for a month in vitro. Subsequently, sustained viral inhibition in bystander Jurkat T cells was observed upon co- culture with NP-exposed CD169+ macrophages. Additionally, our findings suggest that the GM3 lipid coating confers “stealth” properties to suppress non-specific immune activation induced by loaded ARVs and nanoparticle core. Thus, in aim 1, we will investigate the extracellular drug release mechanism from CD169+ macrophages to CD4+ T cells. In aim 2, we will assess the long-term toxicity and stealth mechanism of GM3- PLA-NPs. Finally, we will confirm these results in vivo by assessing the spatial distribution and stealth phenotype of GM3-PLA-NPs in SLTs in aim 3. These studies will elucidate a new approach for safe and effective delivery of long-acting therapeutics to SLTs which could be beneficial for HIV prevention and treatment as well as other diseases such as metastatic cancer.

项目总结/摘要 尽管联合抗逆转录病毒疗法（cART）取得了巨大进展，但HIV仍然是一个全球性威胁。 慢性炎症和免疫激活支持cART中非AIDS共病的风险增加- 治疗艾滋病患者。这种情况归因于各种因素，包括抗逆转录病毒（ARV）毒性和 病毒在次级淋巴组织（SLT）中的持续存在与这些组织中ARV渗透率低有关， 水库地点。此外，每日服用抗逆转录病毒药物仍然是一种负担， 这可能导致病毒从SLTS反弹。在这个项目中，我们建议开发一种长效的 由脂质包裹的聚乳酸纳米颗粒（GM 3-PLA-NP）负载的“隐形”纳米制剂 与非核苷类逆转录酶抑制剂阿匹韦林（RPV）和整合酶抑制剂卡博特拉韦 （CAB）。这些GM 3-PLA-NP具有创新的靶向和设计特征，这将增加药物进入， 保留在SLTs，除了调节炎症反应。事实上， 单唾液酸二己糖神经节苷脂（GM 3）能够特异性结合唾液酸凝集素受体CD 169 骨髓细胞（巨噬细胞和树突细胞），导致非酸性膜的形成 内陷，这可以防止NP降解，并导致细胞药物库的长期建立。 这些NP收集隔室（NPCC）与先前定义的含有病毒的NPCC相同。 CD 169捕获HIV-1后形成的隔室（VCC）。由于VCC也参与骨髓细胞- 通过病毒突触介导的HIV向CD 4 + T细胞的传播（反式感染），我假设， NPCCs可能通过巨噬细胞- T细胞介导ARV向旁观者T细胞的持续释放 突触连接我们的初步数据表明，这些GM 3-PLA-NP成功地被用于治疗糖尿病。 随着时间的推移，在CD 169+巨噬细胞的NPCC中保存，这延长了这些细胞中的抗病毒效力， 体外一个月。随后，观察到旁观者Jurkat T细胞中的持续病毒抑制作用， 用NP暴露的CD 169+巨噬细胞培养。此外，我们的研究结果表明，GM 3脂质涂层 赋予“隐形”特性以抑制由装载的ARV诱导的非特异性免疫活化， 纳米颗粒核心。因此，在目的1中，我们将研究CD 169+细胞的细胞外药物释放机制， 巨噬细胞转化为CD 4 + T细胞。在目标2中，我们将评估GM 3的长期毒性和隐形机制。 PLA-NP。最后，我们将通过评估空间分布和隐身性来证实这些结果 目的3中的SLTs中GM 3-PLA-NP的表型。这些研究将阐明一种新的方法， 有效地将长效治疗剂输送到SLTs，这可能有利于艾滋病毒预防， 治疗以及其他疾病，如转移性癌症。