Defining the impact of cannabinoids on the HIV reservoir in humanized mice

确定大麻素对人源化小鼠 HIV 储存库的影响

• 批准号: 10814024
• 负责人: Edward P Browne
• 金额: $ 53.47万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10814024
• 关键词: ATAC-seq; Acute; Address; Affect; Agonist; Animal Model; Anti-Inflammatory Agents; Biological Assay; Biology; Blood; Brain; CD4 Positive T Lymphocytes; Cannabinoids; Cannabis; Cells; Characteristics; Chronic; Clinical; Data; Dose; Effector Cell; Event; Exhibits; Exposure to; Genetic Transcription; Goals; HIV; HIV Infections; Heterogeneity; Homeostasis; Human; Immune; Immune response; Immune system; Immunity; Immunologic Markers; Immunologics; Individual; Infection; Inflammation; Inflammatory; Learning; Maintenance; Mediating; Methods; Microglia; Modeling; Molecular; Mus; NF-kappa B; Natural Killer Cells; Outcome; Participant; Pathway interactions; Peripheral; Persons; Phase; Phenotype; Population; Research; Signal Induction; Signal Pathway; Solid; Spleen; T-Lymphocyte; THC exposure; Testing; Tissue Sample; Tissues; Viral; Viral Physiology; Viral reservoir; Viremia; Virus Latency; Work; antiretroviral therapy; brain tissue; cannabinoid receptor; cytokine; exhaustion; human data; human tissue; humanized mouse; immunoregulation; in vivo; innovation; insight; latent HIV reservoir; longitudinal human study; marijuana use; marijuana user; molecular phenotype; mouse model; neuroprotection; novel; peripheral blood; polysubstance use; reconstitution; senescence; single cell analysis; tool; transcriptome sequencing; viral DNA; viral RNA

Abstract Cannabis (CB) use is prevalent amongst people with HIV (PWH) but its impact on HIV infection and the latent viral reservoir has not been fully examined. In particular, the impact of CB on the HIV reservoir in solid tissues such as the brain is completely unknown. As such, animal models in which dosing can be carefully controlled and tissue samples can be analyzed will be important to fully understand how CB impacts HIV reservoir characteristics in tissues. Our central hypothesis is that CB induces an immunosuppressive cellular state that impacts viral transcription and maintenance of the HIV reservoir in tissues. The overall goal of this proposal is to develop a humanized mouse model of CB exposure during HIV infection and to use this model to learn how CB use affects characteristics of the tissue resident HIV reservoir. In the R61 phase, HIV infected humanized mice expressing hIL-34 to promote microglial CNS reconstitution will be exposed to a range of THC doses during infection, both before and after viral suppression with antiretroviral therapy (ART). The impact of THC exposure on viremia, viral suppression, immune markers and cytokines in peripheral blood will be examined. In the R33 phase, we will use this model to address three major questions. First, viral DNA (vDNA) and viral RNA (vRNA) levels in the brains and spleens of infected mice that have been dosed with THC will be quantified to determine the impact of THC on the size and transcriptional activity of the viral reservoir in these tissues. Second, we will examine whether THC exposure affects HIV latency reversal in vivo. We hypothesize that cross-talk between inflammatory signaling pathways activated by potent latency reversing agents (LRAs) such as the non-canonical NF-kB agonist AZD5582 and anti-inflammatory pathways activated by CB use could significantly alter the outcome of LRA dosing in vivo. To test this hypothesis, THC or control exposed HIV infected mice will be dosed with AZD5582, and the impact of THC on viremia and viral reactivation in tissues will be examined. Finally, we will use this model to perform a deep single cell characterization of infected cells in the brains and spleens of ART-suppressed humanized mice in the presence and absence of THC exposure. Spleen and brain tissues will be profiled using a combined single cell ATACseq/RNAseq method to identify and characterize HIV vDNA+ or vRNA+ cells and to compare the abundance and phenotype of these cells between CB dosed mice and controls. If successful, this proposal will reveal new insights into how CB affects HIV infection and the characteristics of the tissue-resident HIV reservoir. This in turn will guide novel HIV cure strategies that are optimized for CB-using PWH.

抽象的 大麻（CB）使用在艾滋病毒（PWH）的患者中普遍存在，但对艾滋病毒感染和潜在的影响 病毒储层尚未得到充分检查。特别是，CB对固体组织中HIV储层的影响 例如大脑完全未知。因此，可以仔细控制剂量的动物模型 可以分析组织样品对于充分了解CB如何影响HIV储量非常重要 组织中的特征。我们的中心假设是CB诱导了一种免疫抑制性细胞状态 影响组织中HIV储层的病毒转录和维持。该提议的总体目标是 在HIV感染期间开发人性化的CB暴露鼠标模型，并使用此模型来了解如何 CB使用会影响组织驻留的HIV储藏的特征。在R61阶段，艾滋病毒感染人性化 表达HIL-34以促进小胶质CNS重建的小鼠将暴露于一系列THC剂量 在感染期间，抗逆转录病毒疗法（ART）在病毒抑制之前和之后。 THC的影响 将检查对病毒血症，病毒抑制，免疫标记和细胞因子的暴露。 在R33阶段，我们将使用此模型解决三个主要问题。首先，病毒DNA（VDNA）和病毒 用THC剂量的受感染小鼠的大脑和脾脏中的RNA（VRNA）水平将是 量化以确定THC对病毒储存库的大小和转录活性的影响 组织。其次，我们将检查THC暴露是否会影响体内的HIV潜伏期逆转。我们假设 通过有效潜伏期逆转剂（LRAS）激活的炎症信号通路之间的串扰 例如非典型的NF-KB激动剂AZD5582和CB使用激活的抗炎途径 显着改变了体内LRA剂量的结果。为了检验该假设，THC或对照暴露的HIV 感染的小鼠将用AZD5582剂量，以及THC对组织中病毒血症和病毒重新激活的影响 将被检查。最后，我们将使用该模型执行感染细胞的深层单细胞表征 在THC暴露的情况下，在Art抑制的人源性小鼠的大脑和脾脏中。 脾脏和脑组织将使用组合的单细胞Atacseq/rnaseq方法进行介绍，以识别和 表征HIV VDNA+或VRNA+细胞，并比较这些细胞之间的丰度和表型 CB剂量的小鼠和对照。如果成功，该提案将揭示有关CB如何影响艾滋病毒的新见解 感染和组织居民HIV储藏的特征。反过来，这将指导新颖的艾滋病毒治愈 针对CB使用PWH优化的策略。