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

基本信息

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
项目类别：
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

项目摘要

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) 使用在艾滋病毒感染者 (PWH) 中很普遍，但它对艾滋病毒感染和潜在感染的影响病毒库尚未得到充分检查。特别是，CB 对实体组织中 HIV 储存库的影响比如大脑是完全未知的。因此，可以仔细控制剂量的动物模型可以分析的组织样本对于充分了解CB如何影响HIV储存库非常重要组织中的特征。我们的中心假设是，CB 会诱导一种免疫抑制细胞状态，影响病毒转录和组织中艾滋病毒储存库的维持。该提案的总体目标是开发 HIV 感染期间 CB 暴露的人源化小鼠模型，并使用该模型来了解如何 CB 的使用会影响组织内 HIV 储存库的特征。在R61阶段，HIV感染者人源化表达 hIL-34 以促进小胶质细胞 CNS 重建的小鼠将暴露于一系列 THC 剂量感染期间，包括使用抗逆转录病毒疗法 (ART) 抑制病毒之前和之后。四氢大麻酚的影响将检查外周血中的病毒血症暴露、病毒抑制、免疫标记物和细胞因子。在R33阶段，我们将使用这个模型来解决三个主要问题。一、病毒DNA（vDNA）和病毒服用 THC 的受感染小鼠大脑和脾脏中的 RNA (vRNA) 水平将量化以确定 THC 对这些病毒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 HIV 潜伏期的逆转。我们假设强效潜伏期逆转剂 (LRAs) 激活的炎症信号通路之间的串扰例如非经典 NF-kB 激动剂 AZD5582 和 CB 使用激活的抗炎途径可以显着改变体内 LRA 剂量的结果。为了检验这一假设，THC 或对照暴露的 HIV 受感染的小鼠将接受 AZD5582 给药，以及 THC 对组织中病毒血症和病毒再激活的影响将接受检查。最后，我们将使用该模型对受感染细胞进行深入的单细胞表征在存在或不存在 THC 暴露的情况下，ART 抑制的人源化小鼠的大脑和脾脏中的变化。将使用组合的单细胞 ATACseq/RNAseq 方法对脾脏和脑组织进行分析，以识别和表征 HIV vDNA+ 或 vRNA+ 细胞，并比较这些细胞的丰度和表型 CB 给药小鼠和对照。如果成功，该提案将揭示CB如何影响HIV的新见解感染和组织驻留 HIV 储存库的特征。这反过来又将指导新的艾滋病毒治疗方法针对使用 CB 的 PWH 进行优化的策略。