Impact of Methamphetamine Use on the HIV Nucleome in Individuals on Antiretroviral Therapy

使用甲基苯丙胺对接受抗逆转录病毒治疗的个体 HIV 核组的影响

• 批准号: 9764331
• 负责人: Zhijun Duan
• 金额: $ 101.99万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9764331
• 关键词: 3-Dimensional; Acquired Immunodeficiency Syndrome; Adherence; Affect; Architecture; BACH2 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Culture Techniques; Cell Nucleus; Cell Proliferation; Cell Survival; Cells; ChIP-seq; Chromatin; Chronic; Comorbidity; DNA; DNA biosynthesis; Deoxyribonucleases; Development; Disease Progression; Enhancers; Epigenetic Process; Exposure to; Expression Profiling; Gene Expression; Genes; Genome; Genome engineering; Genomics; Glean; HIV; HIV Infections; HIV resistance; HIV-1; Human; Human Genome; Individual; Inflammation; Insertional Mutagenesis; Investigation; Latent Virus; Lead; Life Expectancy; Location; Maintenance; Maps; Measures; Methamphetamine; Nuclear; Nuclear Structure; Pathology; Patients; Pattern; Phase; Play; Population; Positioning Attribute; Process; Property; Provirus Integration; Proviruses; RNA Splicing; Regulatory Element; Resolution; Risk Factors; Role; STAT5B gene; Site; Structure; Substance Use Disorder; Substance abuse problem; T-Lymphocyte; Techniques; Technology; Treatment Protocols; Viral; Viremia; Virus; Virus Latency; antiretroviral therapy; cell type; cellular engineering; chromatin modification; chromosomal location; cohort; combinatorial; disease transmission; drug of abuse; epigenome; gene function; high risk; immune activation; immune function; in vivo; indexing; insight; integration site; methamphetamine use; novel strategies; pandemic disease; promoter; response; single cell analysis; site-specific integration; structural genomics; transcriptome; transcriptome sequencing; vector

PROJECT SUMMARY/ABSTRACT A major challenge in ending the HIV pandemic is the persistence of the latent HIV reservoir that can lead to viremia, disease progression and transmission to new hosts after discontinuation of antiretroviral therapy (ART). Efficient treatment regimens that successfully eliminate cell populations carrying intact proviruses are not available due to the incomplete understanding of the cellular mechanisms that allow the virus to remain quiescent within the host genome. A compounding risk factor in HIV pathology is substance use disorder (SUD) known to contribute to decreased adherence to and delay of viral decay due to ART, and accelerated progression to AIDS. We hypothesize that persistence of HIV reservoirs is encoded in the proviral location within the 3D architecture of the host genome (nucleome), and influenced by SUD-induced changes in epigenetic structures. During the exploratory high-risk R61 phase of this project, we will first identify SUD-related differences in HIV integration site patterns, the epigenome and the nucleome of CD4+ T lymphocytes derived from patient cohorts that differ in their exposure to methamphetamine (meth). In the second aim, we will apply advanced 3D nucleome mapping strategies to define the interdependence of nuclear structure, proviral genomic location, regulatory elements, and transcriptomes in short-term ex vivo cultivated patient-derived cells from both cohorts. In the third aim, we will use gene-editing techniques to precisely define a role for proviral integration sites (identified in aims 1 and 2) in cellular gene activity and HIV persistence, including those related to meth use. In the R33 phase we propose to assess the impact of targeted provirus integration on nucleome architecture and transcriptomes to clearly identify genes essential for establishing the ART-resistant HIV reservoir. In addition, we will pursue a strategy to define chromatin interactions of proviral integrations and host genes in vivo in human CD4+ T cell populations of HIV-infected individuals.

项目摘要/摘要 结束艾滋病毒大流行的一个主要挑战是潜伏的艾滋病毒蓄水池的持续存在，这可能导致 停止抗逆转录病毒治疗后病毒血症、疾病进展和向新宿主的传播 (艺术)。成功消除携带完整前病毒的细胞群的有效治疗方案是 由于对允许病毒持续存在的细胞机制的不完全了解而无法获得 在宿主基因组中处于静止状态。HIV病理中的一个复合危险因素是物质使用障碍 (SUD)已知有助于由于抗逆转录病毒疗法而减少对病毒的黏附和延缓病毒腐烂，并加速 进展为艾滋病。我们假设HIV宿主的持久性是在前病毒位置编码的 在宿主基因组(核组)的3D架构内，并受SUD诱导的 表观遗传结构。 在该项目的探索性高风险R61阶段，我们将首先确定与艾滋病病毒有关的sod相关差异。 患者外周血中CD_4~+T淋巴细胞的整合部位、表观基因组和核组 接触甲基苯丙胺(冰毒)不同的队列。在第二个目标中，我们将应用先进的 确定核结构、前病毒基因组相互依赖关系的三维核组作图策略 短期体外培养的患者来源的细胞的位置、调控元件和转录本 两组都是。在第三个目标中，我们将使用基因编辑技术来精确定义前病毒的角色 细胞基因活性和艾滋病毒持久性的整合位点(在目标1和2中确定)，包括 与吸食冰毒有关。 在R33阶段，我们建议评估靶向前病毒整合对核组结构的影响 和转录本，以清楚地识别建立抗ART艾滋病毒储存库所必需的基因。在……里面 此外，我们将采用一种策略来定义前病毒整合和宿主基因的染色质相互作用。 活体内人类CD4+T细胞群中的HIV感染者。