T-cell-mediated targeting of therapeutics to HIV reservoirs

描述（由申请人提供）：在发达国家，随着抗逆转录病毒疗法（ART）的出现，艾滋病毒感染者的预后有所改善。然而，抗逆转录病毒疗法既不能治愈感染，也不能完全恢复健康，部分原因是艾滋病毒在组织保护区不断复制。对SIV感染的恒河猴和HIV感染的人类的研究都表明淋巴组织--特别是淋巴结--是进行中复制的重要部位。 控制药物生物分布的因素是复杂的和多因素的，我们认为游离（未包封）药物有效地递送到潜在的HIV复制的所有位点， 是一个巨大的挑战。纳米颗粒（NP）包裹药物的递送是一种替代方案，但在这里，网状内皮系统对NP药物载体的有效清除 可能会干扰分娩我们建议在这项R 01研究中采用我们以前开发的将NP共价连接到T细胞的技术，从而将它们转化为ARV药物货物的活载体，我们称之为T-Pharmacytes（TPH）。我们的方法的一个独特方面是NPs在细胞表面停留数周，并且不干扰T细胞运输或效应子功能。在我们的第一个目标中，我们将NPs与大量CD 4+或CD 8 + T细胞缀合，并利用这些细胞自然归巢到淋巴组织的优势，将ARV递送到NOD/SCID BLT小鼠模型中的这些HIV避难所位点。在一种潜在的非常强大的方法中，我们的第二个目标将是将NP直接连接到已经从HIV感染的受试者中分离的充分表征的HIV特异性CTL克隆，并在huCD 4 + NSG小鼠模型中测试这些克隆。在后一种方法中，我们将用ARV和IL-15超激动剂（IL-15 SA）加载NP。后者都将支持过继转移的细胞的体内存活。 细胞，并将作为潜伏期逆转药物（LRD）。我们设想了一系列的事件：（1）HIV特异性TPH将进入HIV保护区（2）TPH将遇到同源抗原，导致生产性感染细胞的裂解和TPH的局部积累（3）IL-15 SA 并且ARV递送将集中在这些位点，导致i）诱导局部潜伏感染细胞的HIV表达和ii）抑制任何新一轮的感染（4）HIV特异性TPH（通过IL-15 SA增强功能和存活）将识别并消除暴露的潜伏感染细胞。我们的第三个目的是寻求建立在初步研究的基础上，以直接将大量或HIV特异性T细胞与体内NP缀合，从而否定对细胞的离体操作的需要。这种方法将由一个多学科团队开发和测试，该团队将在纳米技术/细胞治疗，HIV T细胞免疫学和HIV感染的人源化小鼠模型方面提供专业知识。最终，我们的研究结果将提供深入了解淋巴组织病毒储库的相对重要性，整体持久性，并可能导致治疗方法，包括持续抑制，或根除，艾滋病毒水库从抗逆转录病毒治疗的个人。

DESCRIPTION (provided by applicant): In developed nations, the prognosis for people living with HIV has improved with the advent of anti-retroviral therapy (ART). However, ART is neither able to cure infection nor to fully restore health due, in part, to ongoing HIV replication in tisses sanctuaries. Studies in both the SIV-infected rhesus macaque and in HIV-infected humans have implicated lymphoid tissue - lymph nodes in particular - as important sites of ongoing replication. The factors governing bio-distribution of drugs are complex and multifactorial and we perceive the effective delivery of free (non-encapsulated) drug to all sites of potential HIV replication to represent a formidable challenge. The delivery of nanoparticle (NP) encapsulated drugs is an alternative, but here the efficient clearance of NP drug carriers by the reticuloendothelial system can be expected to interfere with delivery. We propose in this R01 study to adapt a technology that we have previously developed to covalently attach NPs to T-cells, thus converting them into living carriers of ARV drug cargo that we term T-Pharmacytes (TPH). A unique aspect of our approach is that NPs remain at the cell surface for weeks, and do not interfere with T-cell trafficking or effector functions. In our first Aim we will conjugate NPs to bulk CD4+ or CD8+ T-cells and take advantage of the natural homing of these cells to the lymphoid tissues to deliver ARVs to these HIV sanctuary sites in the NOD/SCID BLT mouse model. In a potentially very powerful approach, our second Aim will be to directly link NPs to well-characterized HIV-specific CTL clones that have been isolated from HIV- infected subjects, and test these in a huCD4+ NSG mouse model. In this latter approach, we will load NPs with both ARVs and an IL-15 superagonist (IL-15SA). The latter will both support the in vivo survival of adoptively transferred cells, and will act as a latency-reversing drug (LRD). We envision a chain of events whereby: (1) HIV-specific TPH will traffic into HIV sanctuary sites (2) TPH will encounter cognate antigen resulting in the lysis of productively infected cells and the local accumulation of TPH (3) IL-15SA and ARVs delivery will be concentrated at these sites resulting in i) induction of HIV expression from local latently-infected cells and ii) suppression of any new rounds of infection (4) HIV-specific TPH (enhanced in function and survival by IL-15SA) will recognize and eliminate exposed latently-infected cells. Our third Aim seeks to build upon preliminary studies to directly conjugate bulk or HIV-specific T-cells to NPs in vivo thus negating the need for ex vivo manipulations of cells. This approach will be developed and tested by a multidisciplinary team bringing expertise in nanotechnology/cell therapy, HIV T-cell immunology, and humanized mouse models of HIV infection. Ultimately our results will provide insights into the relative importance of lymphoid tissue viral reservoirs to overall persistence and may lead to therapeutic approaches comprising either the ongoing suppression of, or the eradication of, HIV reservoirs from ARV-treated individuals.