DNA Methylation as an Epigenetic Host Factor in HIV/AIDS

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已在许多疾病模型（特别是衰老和癌症）中得到广泛研究。这项基于人群的多学科研究的长期目标是确定人类免疫缺陷病毒1型（HIV-1）感染如何改变13个多态性和/或甲基化敏感位点的CpG甲基化谱。在HIV-1传播、发病和耐药相关基因表达的背景下，本研究旨在通过系统分析编码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位点的CpG甲基化模式来验证四个假设；2)关键的免疫调节因子包括干扰素γ、白细胞介素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），负责药物代谢和转运。我们的工作重点是来自“追求卓越的青少年护理和健康”（REACH）队列和赞比亚-乌干达艾滋病研究项目（ZUHRP）的非洲血统受试者。将选择具有纵向临床资料和生物标本（血清、血浆、DNA和细胞）的参与者进行三种主要比较：i) HIV-1感染前与感染后的80-100名ZUHRP血清转化者，ii)有效抗逆转录病毒治疗（ART）前与有效抗逆转录病毒治疗（ART）后的80-100名REACH血清阳性患者，以及iii) HIV-1血清阳性和无ART的REACH和ZUHRP控制者（n=80-100），其血浆病毒载量（VL）最低，而非控制者（n=80-100）血浆病毒载量（VL）最高（按年龄、性别、队列和种族匹配）。分子技术的组合将用于对遗传和表观遗传多态性进行分类；基因表达（mRNA产生和/或蛋白质分泌）将通过逆转录聚合酶链反应（PCR）和酶联免疫吸附试验（ELISA）进行量化。在这些队列中早期发现的其他免疫遗传因素将作为协变量。统计模型将探索个体以及遗传和表观遗传变异的相互作用。总的来说，从这项研究中获得的数据应该允许对复杂和进化的病毒-宿主相互作用进行全面的解剖，并最终增强基于现有的和经常临床测试的拮抗剂、抑制剂、单克隆抗体、质粒构建物和重组产物的干预设计。

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.