T-cell-mediated targeting of therapeutics to HIV reservoirs

描述(申请人提供)：在发达国家，随着抗逆转录病毒疗法(ART)的出现，艾滋病毒携带者的预后有所改善。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，部分原因是组织庇护所正在进行艾滋病毒复制。对感染SIV的恒河猴和感染HIV的人类的研究都表明，淋巴组织--尤其是淋巴结节--是正在进行的复制的重要部位。 控制药物生物分布的因素是复杂的和多因素的，我们认为有效地将免费(非胶囊)药物输送到所有潜在的艾滋病毒复制部位 代表着一个巨大的挑战。纳米粒(NP)包裹药物的递送是一种替代方案，但在这里，通过网状内皮系统有效地清除NP药物载体 可以预期会干扰交付。在这项R01研究中，我们建议采用我们之前开发的一种技术，将NPs共价连接到T细胞上，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)。我们方法的一个独特方面是，NPs在细胞表面停留数周，不会干扰T细胞的运输或效应器功能。在我们的第一个目标中，我们将NPs与大量的CD4+或CD8+T细胞结合，并利用这些细胞自然归巢到淋巴组织的优势，在NOD/SCID BLT小鼠模型中将ARV运送到这些HIV避难所。在一个潜在的非常强大的方法中，我们的第二个目标将是将NPs直接链接到从HIV感染者身上分离出来的具有良好特征的HIV特异性CTL克隆，并在huCD4+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超级激动剂(IL-15SA)负载NPs。后者都将支持过继移植的体内存活。 细胞，并将作为潜伏期逆转药物(LRD)。我们设想了一系列事件，其中：(1)HIV特异性TPH将进入HIV避难所(2)TPH将遇到同源抗原，导致生产性感染细胞的裂解和TPH的局部积聚(3)IL-15SA 抗逆转录病毒药物的传递将集中在这些部位，从而导致：1)诱导局部潜伏感染细胞的艾滋病毒表达；2)抑制任何新一轮感染；4)艾滋病毒特异性TPH(通过IL-15SA增强功能和存活率)将识别并消除暴露的潜伏感染细胞。我们的第三个目标寻求建立在初步研究的基础上，直接将大量的或HIV特异性的T细胞与体内的NPs结合，从而消除体外操作细胞的需要。这一方法将由一个多学科团队开发和测试，该团队将带来纳米技术/细胞疗法、艾滋病毒T细胞免疫学和人源化艾滋病毒感染小鼠模型方面的专业知识。最终，我们的结果将为淋巴组织病毒库对总体持久性的相对重要性提供见解，并可能导致治疗方法，包括正在进行的抑制或根除接受ARV治疗的个体的HIV病毒库。

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.