In vivo study of THC-induced immunogenome changes at single cell resolution in HIV-infected humans

摘要 免疫激活是慢性HIV感染的标志，与合并症风险增加相关， mortality.大麻素（CAN）的先前研究表明，CAN在HIV感染者中具有免疫调节作用。 未受感染的人群。鉴于慢性HIV感染者和CAN患者的炎症状态升高， 可能的抗炎作用，有人认为CAN可以减少HIV疾病的炎症。 然而，CAN在HIV感染情况下减少炎症的能力的研究是矛盾的， CAN在HIV感染环境中的作用的潜在机制仍然知之甚少。我们的初步 人体内施用Δ-9四氢大麻酚（THC）的数据，CAN的主要活性成分， 证明CAN的免疫调节作用因细胞类型而异。因此，了解 THC在特定细胞类型中感染HIV的精确机制。在本申请中，我们假设THC 以细胞类型特异性的方式改变免疫基因组，并通过表观遗传调节改变细胞因子的产生， 这些改变在HIV感染和未感染的宿主基因组之间是不同的。测试 基于这些假设，我们建议在单细胞分辨率下定义表观基因组学和转录组学改变， 在外周血单核细胞中，通过向有和没有HIV感染的人施用THC。 具体而言，我们将1）确定免疫基因和血清细胞因子上的细胞类型特异性mRNA改变 HIV感染者和未感染者中急性THC的改变; 2）定义潜在的表观遗传学 负责急性THC调节基因表达的机制; 3）鉴定表观遗传机制 调节基因表达和血清细胞因子水平的慢性大麻使用在艾滋病毒感染的男性和女性。 使用良好控制的THC挑战范例和严格的研究设计（即，体内和体外，体内和体外， 受试者之间，功能分析，包括两种性别，评价药物的急性和慢性效应）， 我们希望能鉴定出细胞类型特异性基因，其表达受THC影响，并与THC相关。 染色质和DNA甲基化修饰导致细胞因子改变作为功能结果。的 该项目将填补我们对THC对免疫和炎症功能影响的理解方面的知识空白， 体内变化是否与体外观察到的变化一致，以及这些变化是否依赖于HIV status.深入了解基因调控机制，可能导致细胞类型和细胞因子 异常将提供新的见解，CAN是否可以防止炎症性疾病的进展， 艾滋病毒感染者和告知治疗方案。

ABSTRACT Immune activation is a hallmark of chronic HIV infection that is associated with increased risk of comorbidity and mortality. Prior studies of cannabinoids (CAN) show that CAN has immunomodulatory effects in the HIV- uninfected population. Given the elevated inflammatory state in chronic HIV-infected individuals and CAN’s possible anti-inflammatory effects, it has been suggested that CAN may reduce inflammation in HIV disease. However, studies of CAN’s ability to reduce inflammation in the setting of HIV-infection are contradictory and the underlying mechanism of CAN’s effects in the setting of HIV infection remains poorly understood. Our preliminary data in vivo in humans administered Δ-9 tetrahydrocannabinol (THC), the principal active constituent of CAN, demonstrate that the immunomodulatory effects of CAN differ by cell type. Thus, it is critical to understand the precise mechanisms of THC in HIV infection in specific cell types. In this application, we hypothesize that THC alters the immunogenome in a cell type-specific fashion and alters cytokine production via epigenetic regulatory mechanisms and that these alterations differ between HIV-infected and HIV-uninfected host genomes. To test these hypotheses, we propose defining the epigenomic and transcriptomic alterations at single cell resolution in peripheral blood mononuclear cells by administering THC to humans with and without HIV infection. Specifically, we will 1) identify the cell type-specific mRNA alterations on immune genes and serum cytokine alterations by acute THC in HIV-infected and HIV-uninfected individuals; 2) define the underlying epigenetic mechanisms responsible for acute THC-modulated gene expression; 3) identify epigenetic mechanisms regulating gene expression and serum cytokine levels for chronic cannabis use in HIV-infected men and women. Using a well-controlled THC challenge paradigm and rigorous study design (i.e. in vivo and in vitro, within and between subjects, functional profiling, included both sexes, evaluation of acute and chronic effects of the drug), we expect to identify cell-type specific genes whose expression is affected by THC and are associated with chromatin and DNA methylation modifications that lead to cytokine alteration as a functional outcome. The project will fill knowledge gaps in our understanding of THC’s effect on immune and inflammatory functions, whether in vivo changes are consistent with those observed in vitro, and whether these changes depend on HIV status. The in-depth understanding of gene regulatory mechanisms that may result in cell type and cytokine abnormalities will provide novel insights on whether CAN protects against inflammatory disease progression in HIV-infected individuals and inform treatment options.