Transcriptional regulatory mechanisms shaping HIV proviral fate

影响HIV原病毒命运的转录调控机制

基本信息

批准号：
10721394
负责人：
Ivan D'Orso
金额：
$ 5.03万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
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：
2015-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10721394
关键词：
Achievement Applications Grants Architecture Attention Binding Biology Biomedical Research C-terminal CD4 Positive T Lymphocytes Cells Central Nervous System Chromatin Clinical Clonal Expansion Communication Complex Cues Data Epidemic Exposure to Feedback Functional disorder Genetic Transcription Genome Goals HIV HIV Infections Heterogeneity Histone Acetylation Histone Code Immune Immune signaling Immunologic Stimulation In Vitro Infection Integration Host Factors Intervention Knowledge Lymphocyte Maintenance Mediating Mission Modeling Molecular Molecular Probes N-terminal National Institute of Allergy and Infectious Disease Outcome Output Patients Phase Physiological Proviruses Publishing Reader Repression Rest Role Route Sampling Shapes Signal Transduction System Testing Therapeutic Tissues Transcription Elongation Transcription Initiation Transcriptional Activation Transcriptional Regulation Translating Ubiquitin C Ubiquitination Viral Virus Replication Work antiretroviral therapy bench to bedside cell type clinically relevant cofactor genetic corepressor innovation multicatalytic endopeptidase complex new technology novel novel strategies pre-clinical programs promoter reactivation from latency response scaffold targeted treatment therapeutic target ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Over the past two decades, one of the most exciting breakthroughs in biomedical research was the discovery of anti-retroviral therapy (ART), which curbs active HIV replication to nearly undetectable levels. Despite this great achievement, ART fails to cure HIV infection because latent reservoirs persist in tissues containing resting CD4 lymphocytes and other cell types that express low levels of viral products and can be reactivated in response to immune stimulation. Thus, these latent reservoirs are a critical barrier for a functional cure, making the need for new approaches even more urgent. To achieve this major goal, we must first identify novel, critical host factors that can be targeted therapeutically before we can leverage this knowledge for clinical intervention. To this end, this proposal will investigate a master host factor (KAP1) implicated in transducing a variety of physiologic immune signal inputs into proviral transcriptional outputs to reveal the molecular mechanisms and evaluate therapeutic potential. Our work is different from previous studies because it will help uncouple the contribution of the three major features influencing proviral transcription and fate (complex transcriptional circuit architecture, integration landscape, and immune cell state) to help devise rationale approaches for HIV eradication. We hypothesize that both the integration landscape and immune cell state regulate the outcome of an infection (active, latent, reactivated) through the function of critical master host factors like KAP1. New technologies and experimental approaches for investigating this problem provide unique opportunities to test this hypothesis and define the underlying molecular mechanisms. Specifically, we will investigate: 1) how KAP1 communicates with other host co-factors to assemble a positive transcription complex to promote the latency-reactivation switch in response to immune stimulation, 2) how the integration landscape and cell state influence KAP1-mediated proviral transcription activation and clonal expansion, 3) how KAP1 recognizes proviral chromatin to function as a ‘scaffold’ to couple transcription initiation and elongation, and 4) how KAP1 enzymatic functions help relieve the repression imparted by novel transcriptional co-repressors assembled at the provirus during the latency-reactivation switch. The knowledge generated in this study will establish the groundwork for interfering with the function of various “druggable pockets” (e.g., enzymatic and chromatin reader domains) for HIV eradication strategies. The combination of high significance, innovation and use of a diverse set of experimental approaches in physiologically relevant systems (primary models of latency and patient samples) make this proposal unique. Collectively, these studies could have a revolutionary impact on our understanding of HIV latency biology and have therapeutic implications.

项目概要在过去的二十年中，生物医学研究中最令人兴奋的突破之一是发现尽管如此，抗逆转录病毒疗法（ART）仍将活跃的艾滋病毒复制抑制到几乎无法检测的水平。伟大的成就，ART 未能治愈 HIV 感染，因为潜伏病毒库持续存在于含有静息 CD4 淋巴细胞和其他表达低水平病毒产物并可重新激活的细胞类型因此，这些潜在的储存库是功能性治愈的关键屏障，为了实现这一主要目标，我们必须首先确定新方法的必要性。在我们利用这些知识进行治疗之前，可以针对新的、关键的宿主因素进行治疗为此，本提案将调查与此相关的主宿主因子（KAP1）。将各种生理免疫信号输入转变成原病毒转录输出，以揭示我们的工作与以前的研究不同，因为它将有助于解开影响原病毒转录和命运的三个主要特征的贡献（复杂的转录电路架构、集成景观和免疫细胞状态）帮助设计我们了解了整合环境和免疫细胞的基本原理。状态通过关键主机的功能调节感染的结果（主动、潜伏、重新激活） KAP1 等宿主因素为研究这个问题提供了新技术和实验方法。具体来说，我们有独特的机会来检验这一假设并定义潜在的分子机制。将研究：1) KAP1 如何与其他宿主辅助因子通讯以组装正转录促进潜伏期再激活开关响应免疫刺激的复合物，2）如何整合景观和细胞状态影响 KAP1 介导的原病毒转录激活和克隆扩增，3) KAP1 如何识别原病毒染色质作为“支架”来耦合转录起始和延伸，以及 4) KAP1 酶功能如何帮助缓解新型转录所带来的抑制在潜伏期重新激活开关期间在原病毒处组装的共阻遏物。这项研究将为干扰各种“可成药口袋”的功能奠定基础（例如，酶和染色质阅读器域）对于艾滋病毒根除策略的组合具有高度意义，在生理相关系统（主要潜伏期模型和患者样本）使该提议独一无二。总的来说，这些研究可能具有独特性。对我们对艾滋病毒潜伏生物学的理解产生了革命性的影响，并具有治疗意义。