Molecular Imaging of persistent HIV: CD30

项目摘要：根除艾滋病毒的一个主要障碍是存在持续存在的感染细胞 尽管进行了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，许多具有携带艾滋病毒能力的解剖和淋巴隔间是 无法进行常规采样。因此，人们对艾滋病毒的组织负担的了解有限 以及艾滋病毒转录和翻译活性的解剖分布。新颖、非侵入性、活体 诸如基于正电子发射断层扫描(PET)的成像方法等方法可以提供一种手段 来想象水库的样子。一种使用放射性标记SIV抗gp120单抗的PET成像方法 抗体(MAb)已被用于评估猕猴的活动性感染，但类似的方法在 目前还没有人类感染的报告。然而，HIV包膜蛋白特异性mAbs的实施是 在抑制ART的背景下，由于HIV蛋白的低表达，可能是具有挑战性的。一个 另一种方法是开发和实施艾滋病毒感染细胞的非病毒生物标记物的示踪剂。 CD30是肿瘤坏死因子受体超家族的成员，在多种肿瘤细胞(如霍奇金细胞)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ART的作用下高度富含CD30+CD4+T细胞，并使用FDA批准的靶向CD30 细胞毒抗体-药物结合物(ADC)体内和体外的布妥昔单抗-维多丁(BV)导致降低 一些人体内的HIV RNA和DNA水平。重要的是，CD30基因在非小细胞肺癌组织中的表达 接受抗逆转录病毒治疗的参与者仅在HIV RNA+细胞中发现。因为缺乏表达 在未感染的细胞上，CD30是转录活跃的HIV感染细胞的诱人的非病毒标志物， 尽管有压抑的艺术，但仍然坚持。因此，我们建议：(1)合成89Zr-DFO-BV；(2)收集InD 启用体外和体内数据，包括使用µPET/CT对小鼠进行全身剂量估算，(3) 开发高效的免疫反应分析以支持临床翻译，(4)验证和记录 现行良好制造规范(CGMP)生产89Zr-DFO-BV和(5)进行人类第一次 在病毒症、ART受试者和非感染对照中使用89Zr-DFO-BV对CD30进行PET/MR成像 确定药物动力学、剂量学、概念验证和安全性。我们假设有放射性标记的BV 将具有抗CD30的活性，并具有良好的剂量学和药代动力学，供人类使用。 最终，表达CD30的CD4+T细胞的PET/MR成像有可能在组织范围内提供 人类免疫缺陷病毒转录活性细胞在ART上的解剖分布。我们的团队已经建立了一个 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 当前正在进行持久化。因此，这项初步研究是可行的，并有能力提供 为非侵入性成像方法奠定坚实的基础，以加强艾滋病毒的根除和治疗努力。

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.