Molecular Imaging of persistent HIV: CD30

项目概要：根除艾滋病病毒的一个主要障碍是感染细胞的存在， 尽管有抑制性抗逆转录病毒治疗（ART）。HIV主要存在于外周循环之外， 因此，许多具有窝藏HIV能力的解剖和淋巴区室， 无法进行常规采样。因此，对艾滋病毒的组织负荷的了解有限 以及HIV转录和翻译活性的解剖学分布。新型、非侵入性、体内 诸如基于正电子发射断层摄影（PET）的成像方法的方法可以提供一种手段 to visualize可视化the reservoir水库.使用放射性标记的SIV抗gp120单克隆抗体的PET成像方法 抗体（mAb）已被应用于评估猕猴的活动性感染，但类似的方法， 人类还没有报道。然而，HIV包膜蛋白特异性mAb的实施是困难的。 由于在抑制性ART的情况下HIV蛋白的低表达，可能具有挑战性。 另一种方法是开发和实施HIV感染细胞的非病毒生物标志物的示踪剂。 CD30是TNF受体超家族的成员，其在各种肿瘤细胞（例如霍奇金淋巴瘤）上上调 淋巴瘤），但在绝大多数健康细胞上不表达。我们最近证明了HIV RNA 在抑制性ART中高度富集CD30 + CD4 + T细胞，并使用FDA批准的 细胞毒性抗体-药物偶联物（ADC）brentuximab-vedotin（BV）在体内和离体导致 一些个体的HIV RNA和DNA水平。重要的是，CD30 mRNA在组织中的表达， 抗逆转录病毒治疗的参与者只在HIV RNA+细胞中发现。因为缺乏表达 在未感染的细胞上，CD30是HIV感染细胞转录活性的一种诱人的非病毒标记物， 因此，我们建议：（1）合成89Zr-DFO-BV，（2）收集IND 实现体外和体内数据，包括使用µ PET/CT估计小鼠全身剂量测定，（3） 开发一种有效的免疫反应性检测方法，以支持临床翻译，（4）记录并记录 89Zr-DFO-BV的现行药品生产质量管理规范（cGMP）生产和（5）进行首次人体试验 在病毒血症、ART受试者和未感染对照中使用89Zr-DFO-BV进行的CD30 PET/MR成像， 确定药代动力学、剂量学、概念验证和安全性。我们假设放射性标记的BV 将具有抗CD 30的活性，并对人使用具有有利的剂量测定和药代动力学。 最终，表达CD30的CD4 + T细胞的PET/MR成像有可能提供组织范围的 HIV转录活性细胞在ART上的解剖学分布。我们的小组已经建立了一个前， 临床和临床PET-MR成像基础设施，并有三项涉及艾滋病毒成像的人体研究 目前，持续性。因此，这项试点研究是可行的，并有能力提供一个 为非侵入性成像方法奠定了坚实的基础，以加强艾滋病毒的根除和治疗工作。

PROJECT SUMMARY: A major hurdle to HIV eradication is the presence of infected cells that persist despite suppressive antiretroviral therapy (ART). HIV largely resides outside of the peripheral circulation, and thus, numerous anatomical and lymphoid compartments that have the capacity to harbor HIV are inaccessible to routine sampling. As a result, there is a limited understanding of the tissue burden of HIV and the anatomical distribution of HIV transcriptional and translational activity. Novel, non-invasive, in vivo methods, such as positron emission tomography (PET)-based imaging approaches may provide a means to visualize the reservoir. A PET-based imaging approach using a radiolabeled SIV anti-gp120 monoclonal antibody (mAb) has been applied to assess active infection in macaques, but similar approaches in humans have not been reported. However, the implementation of HIV envelope protein specific mAbs is likely to be challenging due to low expression of HIV proteins in the setting of suppressive ART. An alternative approach is to develop and implement tracers for non-viral biomarkers of HIV infected cells. CD30 is a member of the TNF receptor superfamily that is upregulated on various tumor cells (e.g. Hodgkin lymphoma) but not expressed on a vast majority of healthy cells. We recently demonstrated that HIV RNA is highly enriched in CD30+CD4+ T cells on suppressive ART, and targeting CD30 using the FDA approved cytotoxic antibody-drug conjugate (ADC) brentuximab-vedotin (BV) in vivo and ex vivo leads to reduced HIV RNA and DNA levels in some individuals. Importantly, CD30 mRNA expression in tissues from antiretroviral treated participants is found exclusively in HIV RNA+ cells. Because of the lack of expression on uninfected cells, CD30 is an enticing non-viral marker of transcriptionally active HIV-infected cells that persist despite suppressive ART. Therefore, we propose to: (1) Synthesize 89Zr-DFO-BV, (2) Collect IND enabling in vitro and in vivo data including estimate whole-body dosimetry in mice using µPET/CT, (3) Develop an efficient immunoreactivity assay to support clinical translation, (4) Validate and document the current Good Manufacturing Practice (cGMP) production of 89Zr-DFO-BV and (5) Conduct first-in-human PET/MR imaging of CD30 using 89Zr-DFO-BV in viremic, subjects under ART and uninfected controls to determine pharmacokinetics, dosimetry, proof of concept and safety. We hypothesize that radiolabeled BV will have activity against CD30 and have favorable dosimetry and pharmacokinetics for human use. Ultimately, PET/MR imaging of CD30 expressing CD4+ T cells has the potential to provide tissue-wide anatomical distribution of HIV transcriptionally active cells on ART. Our group has an established pre- clinical and clinical PET-MR imaging infrastructure and have three human studies involving imaging HIV persistence currently in process. As a result, this pilot study is feasible and has the capacity to provide a rigorous foundation for non-invasive imaging methods to enhance HIV eradication and therapeutic efforts.