DNA Methylation as an Epigenetic Host Factor in HIV/AIDS

DNA 甲基化作为 HIV/AIDS 的表观遗传宿主因子

• 批准号: 6722900
• 负责人: Jianming Tang
• 金额: $ 25.38万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6722900
• 关键词: AIDS; AIDS therapy; African; CpG islands; DNA methylation; chemokine; clinical research; communicable disease transmission; drug metabolism; drug resistance; enzyme linked immunosorbent assay; gene expression; genetic polymorphism; helper T lymphocyte; human immunodeficiency virus 1; human subject; immunogenetics; microarray technology; pathologic process; polymerase chain reaction

DESCRIPTION (provided by applicant): DNA (CpG) methylation as an epigenetic determinant of transcriptional gene silencing has been studied extensively in many disease models (ageing and cancer in particular). The long-term objective of this population-based and multidisciplinary study is to define how infection with human immunodeficiency virus type 1 (HIV-1) alters the CpG methylation profiles at 13 polymorphic and/or methylation-sensitive loci. Within the context of gene expression in relation to HIV-1 transmission, pathogenesis, and drug resistance, the study aims to test four hypotheses through systematic analyses of CpG methylation patterns at the loci encoding 1) HIV-1 receptor CD4, coreceptors CCR5 and CXCR4, and coreceptor ligands RANTES and SDF-1; 2) key immunoregulatory cytokines including interferon gamma, interleukin 2 (IL-2), IL-4, IL-6, IL-10, and tumor necrosis factor (TNF) alpha; 3) thymidine kinase-1 (TK-1) and multidrug resistance glycoprotein (MDR-1) responsible for drug metabolism and transport. Our work will focus on subjects of African ancestry from the Reaching for Excellence in Adolescent Care and Health (REACH) cohort and the Zambia-UAB HIV Research Project (ZUHRP). Participants with longitudinal clinical data and biological specimens (serum, plasma, DNA and cells) will be selected for three major comparisons: i) 80-100 ZUHRP seroconverters before vs. after HIV-1 infection, ii) 80-100 REACH seropositives before vs. after effective antiretroviral therapy (ART), and iii) HIV-1 seropositive and ART-free REACH and ZUHRP controller (n=80-100) with the lowest plasma viral load (VL) vs. non-controllers (n=80-100) with the highest VL (matched by age, sex, cohort, and ethnicity). A combination of molecular techniques will be used to classify both genetic and epigenetic polymorphisms; gene expression (mRNA production and/or protein secretion) will be quantified by reverse transcription polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Other immunogenetic factors identified earlier in these cohorts will serve as covariates. Statistical models will explore individual as well as interactive effects of genetic and epigenetic variants. Collectively, data derived from this study should allow a comprehensive dissection of the complex and evolving virus-host interplays and ultimately enhance the design of interventions based on existing and often clinically tested antagonists, inhibitors, monoclonal antibodies, plasmid constructs, and recombinant products.

描述(由申请人提供)：DNA(CpG)甲基化作为转录基因沉默的表观遗传决定因素，在许多疾病模型(特别是衰老和癌症)中得到了广泛的研究。这项以人群为基础的多学科研究的长期目标是确定感染人类免疫缺陷病毒1型(HIV-1)如何改变13个多态和/或甲基化敏感基因座的CpG甲基化情况。在与HIV-1传播、发病机制和耐药性相关的基因表达的背景下，该研究旨在通过系统分析编码以下四个假设：1)HIV-1受体CD4、辅助受体CCR5和CXCR4以及辅助受体配体RANTES和SDF-1的CpG甲基化模式；2)关键的免疫调节细胞因子，包括干扰素γ、白介素2(IL-2)、IL-4、IL-6、IL-10和肿瘤坏死因子(TNF)α；3)胸苷激酶-1(TK-1)和多药耐药糖蛋白(MDR-1)负责药物代谢和运输。我们的工作将集中在非洲血统的主题上，包括追求卓越青少年护理和健康(REACH)队列和赞比亚-UAB艾滋病毒研究项目(ZUHRP)。具有纵向临床数据和生物标本(血清、血浆、DNA和细胞)的参与者将被选中进行三项主要比较：i)80-100名ZUHRP血清转换者在感染HIV-1之前和之后，ii)80-100人在有效的抗逆转录病毒治疗(ART)之前和之后达到血清阳性，以及iii)HIV-1血清阳性和不含ART的REACH和ZUHRP控制组(n=80-100)血浆病毒载量(VL)最低与非控制组(n=80-100)VL最高(按年龄、性别、队列和种族匹配)。将使用分子技术的组合来对遗传和表观遗传多态进行分类；基因表达(信使核糖核酸的产生和/或蛋白质分泌)将通过逆转录聚合酶链式反应(PCR)和酶联免疫吸附试验(ELISA)进行量化。这些队列中早先确定的其他免疫遗传因素将作为协变量。统计模型将探索遗传和表观遗传变异的个体和交互影响。总的来说，来自这项研究的数据应该允许对复杂和不断演变的病毒-宿主相互作用进行全面的剖析，并最终加强基于现有的和经常经过临床测试的拮抗剂、抑制剂、单抗、质粒构建和重组产品的干预设计。