Cellular Metabolic Reprogramming and Virus Infectivity

细胞代谢重编程和病毒感染性

• 批准号: 10224633
• 负责人: Richard D'Aquila
• 金额: $ 60.48万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-05 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224633
• 关键词: APOCEC3G gene; Active Sites; Address; Animal Model; Biological Models; Blood; Blood Cells; Cell model; Cells; Clinical; Collaborations; Colon; DNA; FRAP1 gene; Female; Goals; HIV; HIV Infections; Helper-Inducer T-Lymphocyte; Human; ImmunoPET; Infection; Intervention; Knowledge; Literature; Macaca; Mediating; Memory; Metabolic; Modeling; Mucous Membrane; Mus; Observational Study; Outcome; Pathogenesis; Pathogenicity; Predisposition; Production; Publishing; Reporting; SIV; Specimen; System; T memory cell; T-Cell Activation; T-Lymphocyte; Testing; Time; Tissue Model; Tissues; Tonsil; Viral; Viral reservoir; Viremia; Virion; Virus; Virus Replication; cell determination; cell type; design; gene therapy; humanized mouse; immune activation; mTOR Inhibitor; mTOR inhibition; mucosal site; reactivation from latency; reproductive tract; tool; viral RNA; viral rebound

Project Summary This project addresses the following hypotheses: 1. Virus will be spontaneously produced after stopping early ART from the earliest-established “reservoir cells” in mucosal tissues (which Project 1 aims to identify). Candidates for the earliest-established reservoir cells in mucosal tissues (follicular helper T (Tfh), central memory T (Tcm), and Th17 cells) will each be studied ex vivo. T cell latency/reactivation models already described in the literature will be adapted and new models developed that use explanted human and macaque colon mucosa, as well as human tonsils. The former aims to study Th17, and the latter Tfh, cells as potential early reservoirs that may initiate rebound after ART. We will also pilot humanized DRAG mice as a model of female reproductive tract (FRT) mucosal Tfh reservoirs, and determine whether the tonsil explants or humanized mice are preferable for further study of Tfh cell-related mechanisms and interventions (in aims 2 and 3). Macaques with ART-suppression starting at day 3 after mucosal SIV infection, and continuing for 6 months before stopping, will also be studied in close collaboration with project 1. An observational study of HIV-infected subjects before and after pausing >1 year-long suppressive ART will be designed and performed (“Clinical Observation of Rebound after pausing ART”, CORA). 2. mTOR-mediated metabolic reprogramming triggered by T cell activation contributes to viremia rebound off-ART. Cellular mechanisms downstream of mTOR activity will be studied using catalytic mTOR inhibitors in ex vivo tissue and animal models, including in SIV-infected macaques suppressed by ART for 6 months before stopping it. Effects on both virus production from reservoir cells after early ART stops, and target cell susceptibility to virus, will be characterized. 3. Limited or absent virion A3G in virions initially-produced after stopping early ART contribute to the pathogenic cascade to viremia rebound. Model systems (ex vivo and/or humanized DRAG mice) will be used to test whether virus reservoir T cells have low A3G levels even before Vif is expressed (contributing to virus infectivity), and if A3G- boosting tool compounds decrease infectivity of virions produced from T cell reservoirs. We will assess if A3G- mediated decreased virus spread adds to mTOR inhibitor-mediated decreases in uninfected target T cell susceptibility to infection, and if changes in A3G's cytoplasmic distribution caused by mTOR inhibition enhance A3G booster-mediated increases in A3G virion packaging.

项目概要 该项目提出以下假设： 1. 提前停止后病毒会自发产生 ART 来自粘膜组织中最早建立的“储库细胞”（项目 1 旨在识别）。 粘膜组织中最早建立的储库细胞的候选者（滤泡辅助 T (Tfh)、中央 记忆 T (Tcm) 和 Th17 细胞）都将在离体 T 细胞潜伏/再激活模型中进行研究。 文献中描述的内容将被改编，并开发出使用人类和猕猴外植体的新模型 前者旨在研究 Th17 细胞，后者旨在研究 Tfh 细胞的潜力。 ART 后可能引发反弹的早期储存库我们还将试验人源化 DRAG 小鼠作为模型。 女性生殖道 (FRT) 粘膜 Tfh 储库，并确定扁桃体外植体或 人源化小鼠更适合进一步研究 Tfh 细胞相关机制和干预措施（目标 2 3) 猕猴在粘膜 SIV 感染后第 3 天开始进行 ART 抑制，并持续 6 天。 停止前几个月，还将与项目 1 密切合作进行研究。 将设计和执行暂停>1年抑制性ART之前和之后的H​​IV感染者 （“暂停 ART 后反弹的临床观察”，CORA 2. mTOR 介导的代谢重编程）。 T 细胞激活引发的病毒血症会在 ART 下游的细胞机制中反弹。 mTOR 活性将使用催化 mTOR 抑制剂在离体组织和动物模型中进行研究，包括 感染 SIV 的猕猴在停止 ART 之前被抑制 6 个月，对两种病毒产生的影响。 早期 ART 停止后的储存细胞和靶细胞对病毒的易感性将得到表征 3. 有限。 或停止早期 ART 后最初产生的病毒粒子中病毒粒子 A3G 的缺失会导致致病级联 模型系统（离体和/或人源化 DRAG 小鼠）将用于测试病毒血症是否反弹。 甚至在 Vif 表达之前，储存 T 细胞的 A3G 水平就较低（有助于病毒感染性），并且如果 A3G- 增强工具化合物可降低 T 细胞储存库产生的病毒粒子的感染性，我们将评估 A3G- 是否有效。 介导的病毒传播减少加剧了 mTOR 抑制剂介导的未感染靶 T 细胞的减少 对感染的易感性，如果 mTOR 抑制引起的 A3G 细胞质分布的变化增强 A3G 增强剂介导的 A3G 病毒颗粒包装增加。