Turning off HIV White Noise: Switching from Long-Lived to Short-Lived Reservoir

关闭艾滋病毒白噪声：从长寿命库切换到短寿命库

• 批准号: 10676478
• 负责人: Elena Martinelli
• 金额: $ 114.57万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-22 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676478
• 关键词: Ablation; Animals; Apoptosis; Apoptotic; Area; Autopsy; BCL1 Oncogene; Binding; Biological Assay; Bypass; CD28 gene; CD4 Positive T Lymphocytes; Cell Death; Cell Separation; Cell model; Cells; Cellular Metabolic Process; Combined Modality Therapy; Control Groups; Critical Pathways; DNA; Data; Dependence; Development; Disease; Effector Cell; Ensure; Event; Flow Cytometry; Frequencies; Gastrointestinal tract structure; Genetic Transcription; HIV; HIV Infections; Heterogeneity; Immune; Immune response; Immunity; ImmunoPET; Immunologic Suppressor Factors; Immunosuppression; In Vitro; Individual; Infection; Interruption; Intrinsic factor; Investigation; Kinetics; Macaca; Maintenance; Measures; Mediating; Memory; Metabolic; Methods; Microscopy; Modeling; Nature; Nivolumab; Noise; PD-1 blockade; PET/CT scan; Pathway interactions; Pharmaceutical Preparations; Phenotype; Positron-Emission Tomography; Predisposition; Property; Proteomics; Proviruses; Receptors, Adrenergic, beta-1; Resistance; Rest; Rhesus; SIV; Sampling; Signal Transduction; Site; Sleep; Stimulus; Systems Biology; T memory cell; T-Cell Activation; T-Cell Proliferation; T-Lymphocyte; Techniques; Technology; Testing; Therapeutic; Tissue Sample; Tissues; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Viral; Viral reservoir; Virus; anti-PD-1; antiretroviral therapy; design; falls; immune function; in vitro Model; in vivo; in vivo evaluation; inhibitor; innovation; long term memory; lymph nodes; metabolomics; multidisciplinary; novel; novel strategies; pro-apoptotic protein; programmed cell death protein 1; programs; reactivation from latency; response; small molecule inhibitor; spatiotemporal; transcriptomics; viral rebound

HIV persists in all treated individuals and in most cases viral rebound occurs promptly upon antiretroviral treatment interruption (ATI). Tissues are major sites of HIV persistence during cART and, notably, the gastrointestinal tract is the first site were rebound is detectable via SIV-end ImmunoPET/CT in a live SIV infected animal upon ATI. However, a lot is still unknown about the mechanisms of viral persistence in tissues. This is mostly because of logistical barriers in sampling deep tissues and the focal nature of HIV/SIV infection. Our team has developed a sampling workflow based on ImmunoPET/CT signal that overcomes these difficulties and allows sampling of areas of “rebound-competent reservoir” in tissues in a live animal. TGF-β is an important immune suppressor factor, which orchestrates tissue immunity. Levels of TGF-β remain elevated in HIV infected individuals even after years of fully suppressive cART and contribute to immune suppression as well as to the development of non-AIDS-related, non-communicable disorders via pro-fibrotic mechanisms. TGF-β inhibits TCR-driven T cell proliferation and the maturation and function of other immune cell subsets. Importantly, TGF-β is currently being used to induce HIV latency in in vitro models with primary T cells. Our preliminary data demonstrate that blocking TGF-β signaling in vivo favors HIV latency reversal especially in tissues. Moreover, we found that TGF-β blockade stimulates SIV-specific immune responses and decreases BCL-2 expression in memory T cells both in vivo and in vitro. These exciting new data support a view of TGF-β as a critical factor in maintaining immune cells into a resting state mostly resistant to apoptosis. Hence, we hypothesize that blocking TGF-β will not only increase the frequency of latency reversal events, but also enhance the elimination of the viral reservoir by increasing its susceptibility to immune and viral-mediated cell death. We will test this hypothesis by focusing on 3 specific aims. Aim 1 will be an investigation of the mechanisms of TGF-β blockade in vivo. We will leverage PET/CT-guided sampling to obtain tissue areas where virus reverses following TGF-β blockade and analyze the cells in these tissues for their transcriptomic profiles in their own microenviroment. In Aim 2, we will dissect the mechanisms of TGF-β blockade ex vivo to understand the pathways dependence of the effect of TGF-β blockade on HIV latency and on the survival program of central memory T cells. Finally, in Aim 3, we will dissect the combination of TGF-β blockade and PD-1 blockade on the differentiation program and apoptosis sensitivity of the cells harboring the viral reservoir in vivo. In conclusion, we designed a comprehensive strategy that will help us understanding the potential of this novel strategy to bring us closer to an HIV cure.

在所有治疗的个体中，艾滋病毒持续存在，在大多数情况下，病毒反弹发生在抗逆转录病毒上。 治疗中断（ATI）。组织是CART期间HIV持久性的主要部位，尤其是 胃肠道是第一个位点，可以通过SIV-END免疫集/CT检测到Rebound 在ATI时感染动物。但是，关于组织中病毒持久性的机制仍然不明。 这主要是由于对深层组织的后勤障碍和HIV/SIV感染的焦点本质。 我们的团队开发了一个基于Immunopet/CT信号的抽样工作流程，它克服了这些 困难，并允许在活体动物中的组织中抽样“反弹能力的储层”区域。 TGF-β是 一个重要的免疫抑制因子，它策划了组织免疫史。 TGF-β的水平保持升高 在艾滋病毒感染的个体中，即使经过多年的完全抑制作用，也会导致免疫抑制作用 以及通过促纤维化机制发展与非AID相关的，非传染性疾病的发展。 TGF-β抑制了TCR驱动的T细胞增殖以及其他免疫细胞亚群的成熟和功能。 重要的是，目前使用TGF-β来诱导与原代T细胞的体外模型中诱导HIV潜伏期。我们的 初步数据表明，在体内阻止TGF-β信号传导有利于HIV潜伏期逆转，尤其是在 组织。此外，我们发现TGF-β封锁刺激SIV特异性免疫反应并下降 在体内和体外，在记忆T细胞中的Bcl-2表达。这些令人兴奋的新数据支持TGF-β的视图 作为将免疫球维持到静止状态的关键因素，主要抗凋亡。因此，我们 假设阻止TGF-β不仅会增加延迟逆转事件的频率，还会增加 通过增加其对免疫和病毒介导的细胞的敏感性来增强病毒储量的消除 死亡。我们将通过重点关注3个特定目标来检验这一假设。 AIM 1将是 TGF-β封锁在体内的机制。我们将利用PET/CT引导的采样来获得组织区域 其中病毒在TGF-β桶后逆转并分析这些转录组中的细胞 剖面有自己的微观效果。在AIM 2中，我们将剖析TGF-β桶的机理 了解TGF-β阻断对HIV潜伏期和生存的效果的途径依赖性 中央记忆T单元的程序。最后，在AIM 3中，我们将剖析TGF-β阻断和 分化程序的PD-1桶和携带病毒晶状体的细胞的凋亡灵敏度 体内。总之，我们设计了一种全面的策略，将帮助我们理解 这种新颖的策略使我们更接近艾滋病毒治愈方法。