Transcriptional regulatory mechanisms shaping HIV proviral fate

影响HIV原病毒命运的转录调控机制

• 批准号: 9926935
• 负责人: Ivan D'Orso
• 金额: $ 47.65万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 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/9926935
• 关键词: Achievement; Applications Grants; Architecture; Attention; Binding; Biology; Biomedical Research; C-terminal; CD4 Positive T Lymphocytes; Cells; Chromatin; Clinical; Clonal Expansion; Complex; Cues; Data; Epidemic; Exposure to; Feedback; Functional disorder; Genetic Transcription; Genome; Goals; HIV; HIV Infections; Heterogeneity; Histone Acetylation; Histone Code; Immune; Immune signaling; Immunization; In Vitro; Infection; Integration Host Factors; Intervention; Knowledge; Lymphocyte; Maintenance; Mediating; Mission; Modeling; Molecular; Molecular Probes; N Domain; N-terminal; National Institute of Allergy and Infectious Disease; Neuraxis; 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; clinical application; clinically relevant; 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

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.

项目摘要 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 抗逆转录病毒疗法（ART），该疗法将主动的HIV复制弯曲到几乎无法检测到的水平。尽管如此 伟大的成就，艺术无法治愈艾滋病毒感染，因为潜在储层持续存在于包含的组织中 静息CD4淋巴细胞和其他表达低水平病毒产物的细胞类型，可以重新激活 响应免疫刺激。这是这些潜在储层是功能治愈的关键障碍， 更加紧迫新方法的需求。为了实现这一主要目标，我们必须首先确定 新颖的关键宿主因素可以在历史上以历史为目标，然后才能利用这些知识 临床干预。为此，该提案将调查实施的总体因素（KAP1） 将各种生理免疫学免疫信号输入到病毒转录输出中，以揭示 分子机制并评估治疗潜力。我们的工作与以前的研究不同，因为 它将有助于取消三个主要特征影响者转录和命运的贡献 （复杂的转录电路结构，集成景观和免疫细胞状态） 根除艾滋病毒的理由方法。我们假设整合景观和免疫细胞都 国家通过关键主的功能调节感染的结果（活跃，潜在，重新激活） KAP1等宿主因素。调查此问题的新技术和实验方法提供 检验该假设并定义潜在的分子机制的独特机会。具体来说，我们 将调查：1）KAP1如何与其他宿主辅助因子进行交流以组装正转录 复合物以响应免疫刺激而促进延迟反应切换，2）积分如何 景观和细胞状态会影响KAP1介导的提供转录激活和克隆扩张，3） KAP1如何识别前病毒染色质作为“脚手架”的作用，以使夫妇转录启动和 伸长率和4）KAP1酶促函数如何有助于缓解新的转录赋予的表达 在延迟反应开关期间，共抑制剂在预科病毒中组装。在 这项研究将为干扰各种“可药袋”的功能建立基础（例如， 用于消除HIV策略的酶和染色质读取器领域。高意义的结合， 在物理相关系统中的一系列实验方法的创新和使用（主要 延迟和患者样本的模型）使该建议与众不同。总的来说，这些研究可能具有 革命性的影响我们对艾滋病毒潜伏生物学的理解并具有治疗意义。