GM3 Nanoparticles for Sustained Delivery of Anti-Retrovirals to Lymphatic Tissues

项目摘要 尽管抗逆转录病毒疗法（ART）的广泛实施，有效地抑制病毒复制， 在外周，HIV持续存在于组织储库中，并导致HIV中的快速血浆病毒反弹- 已停止ART的受感染个体。 代表药物特权位点，可解释接受ART治疗的HIV感染个体中残留病毒的持久性。 因此，有效的治愈和根除策略需要开发新的方法， 目标是淋巴结内的HIV宿主。提高抗逆转录病毒药物的有效策略 在次级淋巴结中的浓度是将ARV配制在载体中，例如纳米颗粒， 化合物通过淋巴管转运至引流淋巴结。此外，预防战略 快速清除并在淋巴结中长时间维持ARV的抑制浓度， 需要有效抑制持续的病毒表达。因此，在本申请中，我们建议 开发含有GM 3的膜包封纳米颗粒的方法，以建立长期的 在淋巴结中的抗逆转录病毒药物的组织库，以抑制残留的病毒复制。这些纳米粒子的设计 是基于我们最近的发现，证明特异性捕获GM 3（α 2，3-连接唾液酸含 单唾液酸-二己糖神经节苷脂）的膜包封的金纳米颗粒通过表达CD 169 原代人骨髓细胞如巨噬细胞和树突状细胞体外和GM 3-金的共定位 小鼠淋巴结中具有CD 169+细胞的NP。有趣的是，GM 3-金纳米颗粒的捕获导致 在表面可及的非溶酶体质膜内陷中的隔离和保存， CD 169+骨髓细胞。因此，我们将利用含GM 3的膜之间的这些特异性相互作用， 包封的纳米颗粒和CD 169，以实现淋巴结驻留骨髓的选择性靶向 细胞和隔离到专门的非溶酶体区室，以形成“药物仓库”， 抗逆转录病毒药物的持续抑制浓度。最后，我们将确定这些新型ARV的能力- 含有纳米载体以抑制HIV-1人源化小鼠模型中淋巴结中的病毒复制 感染我们相信这些研究对于发展病毒根除策略， 持续的HIV储存库。

PROJECT ABSTRACT Despite widespread implementation of antiretroviral therapy (ART) that effectively suppresses virus replication in the periphery, HIV persists in tissue reservoirs and accounts for the rapid plasma viral rebound in HIV- infected individuals who have discontinued ART. It is well appreciated that secondary lymphoid tissues represent a drug-privileged site that account for residual viral persistence in HIV-infected individuals on ART. Hence, effective cure and eradication strategies require development of novel approaches that specifically target lymph node-resident HIV reservoirs. An effective strategy to enhance anti-retroviral (ARV) drug concentrations in secondary lymph nodes is to formulate ARVs in carriers such as nanoparticles that promote transport of compounds via the lymphatics to the draining lymph nodes. Additionally, strategies that prevent rapid clearance and maintain inhibitory concentrations of ARVs in lymph nodes for an extended time are needed to effectively suppress persistent virus expression. Hence in this application, we are proposing to develop GM3-containing membrane encapsulated nanoparticle-based approaches to establish long-term tissue depots of ARVs in lymph nodes to suppress residual viral replication. The design of these nanoparticles is based on our recent findings that demonstrate specific capture of GM3 (α2,3-linked sialic acid containing monosialo-dihexosylganglioside) containing membrane encapsulated gold nanoparticles by CD169 expressing primary human myeloid cells such as macrophages and dendritic cells in vitro and co-localization of GM3-gold NPs with CD169+ cells in lymph nodes of mice. Interestingly, capture of GM3-gold NPs resulted in sequestration and preservation within surface accessible non-lysosomal plasma membrane invaginations in CD169+ myeloid cells. Hence, we will exploit these specific interactions between GM3-containing membrane encapsulated nanoparticles and CD169 to achieve both selective targeting of lymph node resident-myeloid cells and sequestration to specialized non-lysosomal compartments for formation of “drug depots” that achieve sustained inhibitory concentrations of ARVs. Finally, we will determine the ability of these novel ARV- containing nanocarriers to suppress virus replication in lymph nodes in a humanized mouse model of HIV-1 infection. We believe these studies are critical to development of virus-eradication strategies for eliminating persistent HIV reservoir.