The contribution of unintegrated HIV-1 to latency and to models of latency

未整合的 HIV-1 对潜伏期和潜伏期模型的贡献

基本信息

批准号：
9075588
负责人：
DAVID N LEVY
金额：
$ 39.63万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9075588
关键词：
Accounting Acute Antiviral Therapy Binding Proteins CD4 Positive T Lymphocytes Cell Survival Cells Chromatin Competence Constitution DNA DNA Methylation DNA Modification Process Data Development Essential Genes Gene Expression Genetic Transcription Genome Goals HIV HIV Infections HIV-1 Health Heterochromatin In Vitro Infection Integrase Integrase Inhibitors Interleukin-4 Invaded Investigation Methylation Modeling Molecular Mutation Phase Population Process Production Proliferation Marker Proviruses Publishing Regulation Relative (related person)Reporting Resistance Rest Resting Phase Role Series Shock Structure System T-Cell Activation T-Lymphocyte Testing Time Transcript Viral Virus base cell type chromatin modification chromatin remodeling epigenetic regulation histone methylation histone modification in vitro Model in vivo insight killings novel response therapeutic development therapy development viral RNA vpr Genes

项目摘要

DESCRIPTION (provided by applicant): HIV infection remains incurable owing to the persistence of viral reservoirs that are resistant to current antiviral therapies. The latent reseroir is rapidly established during acute/early infection, and during this phase many viral RNA+ cells lack activation and proliferation markers and thus resemble resting CD4+ T cells. These findings suggest that latency is often established by direct infection of resting T cells. In vitro models o HIV-1 latency frequently employ direct infection of resting CD4 T cells, including our recently described latency system utilizing IL-4 to render T cells infectible without inducing T cell activation. We have reported that when HIV-1 infects resting CD4 T cells, large numbers of latently infected cells are generated which contain HIV-1 genomes that have not integrated and cannot integrate, yet these genomes are capable of de novo virus production when the cell is activated several days after infection. Using an IL-4-based latency system, we find that cells containing only unintegrated HIV-1 constitute about one half of all latently infected cells. We fin that latent unintegrated genomes respond to shock and kill agents but that their responses indicate epigenetic regulation that differs from integrated proviruses. We hypothesize that this new form of latency is a common occurrence in standard in vitro models of HIV-1 latency. The studies we propose will define the mechanisms controlling this new form of latency and its contribution to in vitro models of HIV-1 latency. We further find that the Vpr gene of HIV-1 is a key regulator the chromatization of HIV-1 genomes, and we will define the mechanisms by which it performs this function. We find that Vpr is essential for expression from uDNA in resting T cells, and that without Vpr, these genomes are unresponsive to stimulation and display altered chromatin methylation. We hypothesize that Vpr counteracts cellular defenses against invading parasitic DNA that install transcriptionally repressive chromatin modifications. This project will provide new insights into the establishment and regulation of HIV-1 latency. This project will also provide necessary information on the constitution and functioning of important in vitro models of latency that must be accounted for in the development of novel anti-viral therapies.

描述（由适用提供）：由于对当前抗病毒疗法有抵抗力的病毒储量的持续存在，HIV感染仍然无法治愈。在急性/早期感染期间，潜在的reseroir迅速建立，在此阶段，许多病毒RNA+细胞缺乏激活和增殖标记，因此类似于静息CD4+ T细胞。这些发现表明，延迟通常是通过直接感染静息T细胞建立的。体外模型O HIV-1潜伏期经常采用静息CD4 T细胞的直接感染，包括我们最近描述的潜伏系统利用IL-4来使受感染的T细胞感染而无需诱导T细胞激活。我们报道说，当HIV-1感染静止的CD4 T细胞时，会产生大量的潜在感染细胞，其中包含尚未整合且无法整合的HIV-1基因组，但这些基因组在感染几天后能够激活细胞时产生从头病毒。使用基于IL-4的延迟系统，我们发现细胞仅包含未整合的HIV-1构成所有潜在感染细胞的一半。我们认为潜在的不整合基因组对冲击和杀死药物的反应反应，但它们的反应表明表观遗传调节与综合预科病毒不同。我们假设这种新形式的延迟是HIV-1潜伏期的标准体外模型中的常见。我们提出的研究将定义控制这种新形式潜伏期及其对HIV-1潜伏期体外模型的贡献的机制。我们进一步发现，HIV-1的VPR基因是HIV-1基因组色谱的关键调节剂，我们将定义其执行此功能的机制。我们发现VPR对于静息T细胞中UDNA的表达至关重要，并且如果没有VPR，这些基因组对刺激无反应，并且显示出改变的染色质甲基化。我们假设VPR可以抵消细胞防御剂，以防止入侵寄生DNA，该寄生DNA安装了转录反射性染色质修饰。该项目将为HIV-1潜伏期的建立和调节提供新的见解。该项目还将提供有关重要体外潜伏模型的配置和功能的必要信息，这些信息在开发新型抗病毒疗法时必须考虑到。