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.

HIV 在所有接受治疗的个体中持续存在，并且在大多数情况下，抗逆转录病毒治疗后病毒会立即反弹 治疗中断 (ATI) 组织是 cART 期间 HIV 持续存在的主要部位，尤其是治疗中断。 胃肠道是活体 SIV 中通过 SIV 末端免疫 PET/CT 检测到反弹的第一个部位 然而，关于病毒在组织中持续存在的机制仍然有很多未知之处。 这主要是因为深部组织取样的后勤障碍以及 HIV/SIV 感染的焦点性质。 我们的团队开发了基于ImmunoPET/CT信号的采样工作流程，克服了这些问题 困难并允许对活体动物组织中的“具有反弹能力的储存库”区域进行采样。 一种重要的免疫抑制因子，可协调组织免疫 TGF-β 水平保持升高。 即使在经过多年的完全抑制性 cART 治疗后，HIV 感染者仍会出现这种情况，并有助于免疫抑制 以及通过促纤维化机制发展出与艾滋病无关的非传染性疾病。 TGF-β 抑制 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，TGF-β 目前被用于在原代 T 细胞体外模型中诱导 HIV 潜伏期。 初步数据表明，体内阻断 TGF-β 信号传导有利于 HIV 潜伏期逆转，尤其是在 此外，我们发现 TGF-β 阻断会刺激 SIV 特异性免疫反应并降低。 体内和体外记忆 T 细胞中 BCL-2 的表达这些令人兴奋的新数据支持了 TGF-β 的观点。 作为维持免疫细胞进入静息状态的关键因素，主要抵抗细胞凋亡。 坚持认为阻断 TGF-β 不仅会增加潜伏期逆转事件的频率，而且 通过增加病毒库对免疫和病毒介导细胞的易感性来增强病毒库的消除 我们将通过关注 3 个具体目标来检验这一假设。目标 1 将是对死亡的调查。 我们将利用 PET/CT 引导采样来获取组织区域。 病毒在 TGF-β 阻断后逆转并分析这些组织中的细胞的转录组 在目标 2 中，我们将剖析 TGF-β 体外阻断的机制。 了解 TGF-β 阻断对 HIV 潜伏期和存活率影响的途径依赖性 最后，在目标 3 中，我们将剖析 TGF-β 阻断和 PD-1 阻断病毒库细胞的分化程序和凋亡敏感性 总之，我们设计了一个全面的策略，将帮助我们了解体内的潜力。 这种新颖的策略使我们更接近治愈艾滋病毒。