Epigenetic regulation of HIV latency

HIV潜伏期的表观遗传调控

基本信息

批准号：
8500881
负责人：
Eric M. Verdin
金额：
$ 18.39万
依托单位：
J. DAVID GLADSTONE INSTITUTES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8500881
关键词：
Activated Lymphocyte Affect Affinity Chromatography Beryllium Binding Biological Assay Brain CD4 Positive T Lymphocytes Cell Line Cell model Cells Chromatin Cocaine Cocaine Users Collaborations Complementary DNA Complex CpG Islands CpG dinucleotide DNA DNA Methylation DNA Methylation Inhibition Deoxycytidine Drug Delivery Systems Drug usage Dysmyelopoietic Syndromes Epigenetic Process Gene Expression Genes Genetic Transcription Genome Genomics HDAC1 gene HIV HIV-1 Highly Active Antiretroviral Therapy Histone Deacetylation Human Immune system In Vitro Individual Jurkat Cells Latent Virus Lead Length Libraries Link Lymphocyte Lymphocyte Activation Lymphoid Lymphoid Cell Maintenance Mediating Methyl-CpG-Binding Protein 2 Methylation Methyltransferase Modeling Modification Multiprotein Complexes Nucleosomes Patients Pattern Pharmaceutical Preparations Play Process Protein Binding Domain Proteins Proviruses RNA Interference Recombinant Proteins Recruitment Activity Regulation Reporting Repression Rest Role Site Stimulus System T-Cell Activation TNF gene TNFRSF5 gene Testing Transcription Repressor/Corepressor Virus Virus Integration Virus Latency Work antiretroviral therapy base cDNA Library chromatin immunoprecipitation cocaine use drug abuser immune activation in vitro Model inhibitor/antagonist latent infection member new therapeutic target novel novel therapeutic intervention prevent promoter prostratin protein complex public health relevance research study small hairpin RNA small molecule viral DNA

项目摘要

DESCRIPTION (provided by applicant): ABSTRACT The presence of latent reservoirs has prevented the eradication of Human Immunodeficiency Virus (HIV) from infected patients successfully treated with antiretroviral therapy. We have recently reported that DNA methylation of the HIV promoter plays an important role in the establishment and maintenance of HIV latency. We also found that the inhibitor of DNA methylation, 5-aza-2'deoxycytidine (aza-CdR), synergizes with NF-B activators to reactivate latent HIV. We propose to further test the hypothesis that methylation of the HIV promoter plays a critical role in HIV latency and that inhibition of DNA methylation represents a novel therapeutic approach to induce the reactivation of latent HIV. Since cocaine has emerged as a molecule that can modify epigenetic regulatory processes we also plan to examine its effect in HIV latency. Our specific plans are to: Aim 1. To understand the mechanism of HIV promoter silencing via DNA methylation. We will use chromatin immunoprecipitation and shRNA-mediated interference to explore the roles in HIV latency of the multiprotein complexes that are recruited to methylated DNA: NuRD/MBD2, SWI/SNF/Sin3/MeCP2 and the SUV39H1/HP1/MBD1 complexes. Aim 2. To identify small molecules and cellular factors that reactivate latent HIV in synergy with methylation inhibitors. We will screen for small molecules and cDNAs that synergize with aza-CdR to reactivate latent HIV. Agents that synergize with Aza-CdR to reactivate HIV in the J-Lat model of HIV latency will be tested in a primary cell models of latency and in latently infected cells from HIV-positive donors. Aim 3. To study HIV DNA methylation in primary lymphocytes. To test the hypothesis that HIV integration in or near hypermethylated CpG islands leads to latency, we propose to compare the state of DNA methylation in resting and activated lymphocytes at the sites of latent HIV integration in comparison to productive sites both in the J-Lat model and in latent HIV in primary human lymphoid cells. Aim 4. To study the size of the latent pool in HIV-infected patients, its reactivation by methylation inhibitors and the effect of cocaine use on these variables. We propose to test for synergistic reactivation of latent HIV in latently infected cells using a primary lymphoid system for HIV latency in vitro (in collaboration with Dr, Vicente Planelles) and in latently-infected cells isolated from HIV-infected patients both cocaine users and non users. PUBLIC HEALTH RELEVANCE: NARRATIVE The identification of DNA methylation and other epigenetic modifications of the latent HIV genome highlights a novel mechanism for the suppression of HIV transcription. The demonstration that the DNA methylation inhibitor, aza-DcR, synergizes with prostratin or TNF- to reactivate latent HIV suggest that epigenetic modification could be used as a drug target in our effort to reactivate latent HIV. Recent experiments also indicate that cocaine use can induce prolonged epigenetic modifications in the brain and possibly in the immune system. Here we propose to study the role of epigenetic modifications in HIV latency and the possible role of drugs that modify epigenetic silencing as a novel therapeutic approach to induce the reactivation of latent HIV.

描述（由申请人提供）：摘要潜在储层的存在阻止了成功通过抗逆转录病毒治疗的感染患者消除了人类免疫缺陷病毒（HIV）。我们最近报告说，HIV启动子的DNA甲基化在HIV潜伏期的建立和维持中起着重要作用。我们还发现，DNA甲基化的抑制剂，5-Aza-2'deoxyCytidine（AZA-CDR），与NF-B激活剂协同作用，以重新激活潜在的HIV。我们建议进一步检验以下假设：HIV启动子的甲基化在HIV潜伏期中起关键作用，并且抑制DNA甲基化代表了一种新型的治疗方法来诱导潜在HIV的重新激活。由于可卡因已成为可以修改表观遗传调节过程的分子，因此我们还计划检查其在HIV潜伏期中的影响。我们的具体计划是：目标1。了解通过DNA甲基化沉默的HIV启动子的机制。我们将使用染色质免疫沉淀和SHRNA介导的干扰来探索募集到甲基化DNA的多蛋白络合物的艾滋病毒潜伏期中的作用：NURD/MBD2，SWI/SNF/SNF/SIN3/MECP2和SUV39H1/HP1/HP1/MBD1复合物。目的2。鉴定小分子和细胞因子，这些分子与甲基化抑制剂协同作用中的潜在HIV重新激活。我们将筛选与AZA-CDR协同重新激活潜在HIV的小分子和cDNA。在HIV潜伏期的J-LAT模型中与AZA-CDR协同激活HIV的药物将在潜伏期的主要细胞模型和HIV阳性供体的潜在感染细胞中进行测试。目的3。研究原发性淋巴细胞中的HIV DNA甲基化。为了检验以下假设：在高甲基化的CpG岛中或附近的HIV整合会导致潜伏期，我们建议比较潜在HIV整合部位的静息和活化淋巴细胞中的DNA甲基化状态，而不是J-LAT模型中的生产性位点在原发性人淋巴样细胞中的潜在艾滋病毒中。目的4。研究HIV感染患者的潜在池的大小，其通过甲基化抑制剂重新激活以及可卡因使用对这些变量的影响。我们建议使用主要的淋巴机系统（与DR，Vicente Planelles合作）和从HIV感染的患者分离的艾滋病毒潜伏期（与HIV感染的患者分离的艾滋病毒潜伏期）测试潜在感染细胞中潜在HIV的协同重新激活。用户。公共卫生相关性：叙述性鉴定DNA甲基化和潜在HIV基因组的其他表观遗传修饰突出了一种抑制HIV转录的新机制。 DNA甲基化抑制剂AZA-DCR与prostratin或tnf-重新激活潜在的HIV的证明表明，表观遗传修饰可以用作我们努力重新激活潜伏HIV的药物靶标。最近的实验还表明，可卡因的使用可以诱导大脑和免疫系统中的长期表观遗传修饰。在这里，我们建议研究表观遗传修饰在艾滋病毒潜伏期中的作用，以及将表观遗传沉默作为一种新型的治疗方法来诱导潜在艾滋病毒的重新激活的药物的可能作用。