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

摘要 免疫激活是慢性艾滋病毒感染的一个标志，它与共病风险增加和 死亡率。先前对大麻素(CaN)的研究表明，CaN对艾滋病毒具有免疫调节作用。 未受感染的人群。鉴于慢性HIV感染者和CAN的炎症状态升高 可能的抗炎作用，有研究表明，CAN可能会减轻HIV疾病的炎症。 然而，关于CaN在HIV感染环境中减少炎症的能力的研究是相互矛盾的，而且 CaN在HIV感染环境中的作用的潜在机制仍然知之甚少。我们的预赛 在服用Δ-9四氢大麻酚(CaN的主要活性成分)的人体内的数据， 证明免疫调节作用因细胞类型不同而不同。因此，理解 THC在特定细胞类型中感染HIV的精确机制。在本应用程序中，我们假设THC 以细胞类型特异的方式改变免疫原组，并通过表观遗传调节改变细胞因子的产生 这些变化在感染艾滋病毒的宿主基因组和未感染艾滋病毒的宿主基因组之间是不同的。为了测试 这些假说，我们建议在单细胞分辨率下定义表观基因组和转录改变 在外周血单核细胞中，通过给感染和不感染艾滋病毒的人注射THC。 具体地说，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变化。 HIV感染者和非感染者急性THC的改变；2)定义潜在的表观遗传学 急性THC调控基因表达的机制；3)识别表观遗传机制 调节艾滋病毒感染男女慢性使用大麻的基因表达和血清细胞因子水平。 使用控制良好的THC挑战范式和严格的研究设计(即体内和体外、体内和 受试者之间的功能描述，包括性别，药物的急性和慢性影响的评估)， 我们希望确定其表达受THC影响的细胞类型特定基因，并与 染色质和DNA甲基化修饰导致细胞因子改变作为功能结果。这个 该项目将填补我们在了解THC对免疫和炎症功能影响方面的知识空白， 体内的变化是否与体外观察到的一致，以及这些变化是否取决于艾滋病毒 状态。对可能导致细胞类型和细胞因子的基因调控机制的深入了解 异常将提供关于是否可以预防炎症性疾病进展的新见解 为艾滋病毒感染者提供信息，并告知治疗方案。

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.