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

HIV 感染者单细胞分辨率 THC 诱导免疫基因组变化的体内研究

• 批准号: 10682520
• 负责人: KE XU
• 金额: $ 49.16万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10682520
• 关键词: ATAC-seq; Acute; Adaptive Immune System; Affect; Anabolism; Antiinflammatory Effect; Cannabinoids; Cannabis; Cells; Chromatin; Chromatin Structure; Chronic; Complex; DNA Methylation; Data; Disease; Disease Progression; Dose; Epigenetic Process; Evaluation; Gene Expression; Genes; Genetic Transcription; Genome; HIV; HIV Infections; Hour; Human; Immune; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Infusion procedures; Intravenous infusion procedures; Knowledge; Maps; Medical; Messenger RNA; Modeling; Modification; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Play; Population; Production; Publishing; Regulation; Regulator Genes; Reporting; Research Design; Resolution; Risk; Role; S100A8 gene; S100A9 gene; Sampling; Serum; Testing; Tetrahydrocannabinol; Time; Validation; Woman; cell type; cohort; comorbidity; cytokine; differential expression; epigenomics; functional outcomes; immune activation; immune function; immunoregulation; in vivo; innovation; insight; mRNA Expression; marijuana use; men; methylome; mortality; novel; sex; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transcriptomics

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.

抽象的 免疫激活是慢性艾滋病毒感染的标志，与合并症的风险增加有关 死亡。先前对大麻素（CAN）的研究表明，可以在HIV中具有免疫调节作用 未感染的人群。考虑到慢性艾滋病毒感染者的炎症状态升高 可能的抗炎作用，已经提出可以减少艾滋病毒疾病的感染。 但是，关于CAN在HIV感染情况下降低感染能力的研究是矛盾的，并且 在艾滋病毒感染的情况下，CAN的影响的基本机制仍然很少了解。我们的初步 人体中的数据施用Δ-9四氢大麻酚（THC），CAN的主要活性构成， 证明可以通过细胞类型不同的免疫调节作用。那是至关重要的 特定细胞类型中艾滋病毒感染中THC的精确机制。在此应用程序中，我们假设THC 以特定于细胞类型的方式改变免疫原子，并通过表观遗传调节来改变细胞因子的产生 机制和这些改变在感染HIV的和HIV未感染的宿主基因组之间。测试 这些假设，我们提出在单细胞分辨率下定义表观基因组和转录组的变化 在外周血单核细胞中，通过对有或没有HIV感染的人类进行THC。 具体而言，我们将1）确定免疫原和血清细胞因子的细胞类型特异性mRNA改变 急性THC在感染HIV和HIV未感染的个体中的改变； 2）定义潜在的表观遗传 负责急性THC调节基因表达的机制； 3）确定表观遗传机制 调节在HIV感染的男性和女性中使用慢性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，内部和 在受试者之间，功能分析，包括性别，对药物的急性和慢性作用的评估）， 我们期望鉴定细胞类型的特定基因，其表达受到THC的影响并与 导致细胞因子改变作为功能结果的染色质和DNA甲基化修饰。这 项目将填补我们对THC对免疫和炎症功能的影响的理解，以填补知识空白， 体内变化是否与体外观察到的变化一致，以及这些变化是否取决于HIV 地位。对可能导致细胞类型和细胞因子的基因调节机制的深入了解 异常将提供有关是否可以防止炎症性疾病进展的新见解 艾滋病毒感染的人并为治疗方案提供信息。