Transcriptional Crosstalk between DUX4-FL and HIV

DUX4-FL 和 HIV 之间的转录串扰

• 批准号: 10693294
• 负责人: Sonia Mediouni Jablonski
• 金额: $ 28.98万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693294
• 关键词: Acceleration; Binding; Biology; Blood; CD4 Positive T Lymphocytes; Cell Line; Cells; Chromatin; Co-Immunoprecipitations; Color; Complex; Disease; Doxycycline; EP300 gene; Embryonic Development; Encapsulated; Ensure; Epigenetic Process; Evolution; Facioscapulohumeral Muscular Dystrophy; Flow Cytometry; Future; Genes; Genetic Transcription; Genomics; Goals; HIV; HIV Infections; HIV Seronegativity; HIV Seropositivity; Heterochromatin; Histones; Homeodomain Proteins; Human; Immune; Immune Evasion; Immune system; Impairment; In Vitro; Individual; Infection; Internal Ribosome Entry Site; Interphase Cell; Interruption; Jurkat Cells; Knowledge; Label; Laboratories; Length; Leukapheresis; Life; Link; Malignant Neoplasms; Mammalian Cell; Mediating; Methods; Modeling; Mus; Myoblasts; Persons; Pharmaceutical Preparations; Poly(A)+ RNA; Process; Production; Productivity; Protein Isoforms; Proteins; Proteomics; Proviruses; RNA Polymerase II; RNA analysis; Regulation; Relaxation; Reporter; Reporting; Reproducibility; Residual state; Role; Sampling; Sorting; Therapeutic; Toxic effect; Transcript; Transcriptional Elongation Factors; Transcriptional Regulation; Variant; Viral; Viral Proteins; Viremia; antiretroviral therapy; chromatin remodeling; cortistatin; detection limit; digital; disease phenotype; drug development; epigenetic silencing; in vivo Model; inhibitor; insight; knock-down; memory CD4 T lymphocyte; new therapeutic target; overexpression; prevent; promoter; protein complex; reactivation from latency; recruit; tat Protein; transcription factor; transcriptome sequencing; viral RNA; viral rebound

Abstract HIV eradication strategies aim to clear residual HIV transcription, from a pool of latently infected memory CD4+ T cells, that persist in people living with HIV (PLWH) treated with an effective antiretroviral therapy (ART). These cells harbor a provirus poised to viral rebound upon a sequential recruitment, at the HIV promoter, of host transcription elongation factors, the viral Tat protein and host chromatin remodelers (a process called Tat- mediated transcription). Long-term block of this residual HIV transcription, in different models of HIV latency, using an inhibitor of Tat, results in an heterochromatinization and a loss of RNA polymerase II at the HIV promoter, along with preventing viral rebound. Thus, our premise is that transcriptional and chromatin remodeling inhibitors along with Tat inhibitors can be used in the block-and-lock functional cure approaches, aimed at reducing residual viremia during ART and limiting viral rebound. Additionally, gaining fresh insightful knowledge on the connections between these factors and HIV transcription will help leverage them as antiviral targets. We have uncovered a direct positive relationship between the expression of the full-length isoform of the double homeobox protein 4 (DUX4-FL) expression and HIV infection in primary blood CD4+T cells. DUX4-FL robustly activates HIV transcriptional activity by promoting both residual and Tat-mediated HIV transcription. Furthermore, HIV enhances DUX4-FL transcription suggesting a positive transcriptional crosstalk between HIV and DUX4-FL. DUX4-FL is a strong transcription factor that is mostly expressed in disease phenotypes, such as facioscapulohumeral muscular dystrophy and cancers, but has never been described to regulate HIV. In disease, DUX4-FL typically regulates expression of evolutionally or immune adaptative genes, resulting in toxicity and/or evasion from the immune system. DUX4-FL has also been described to open compacted chromatin by recruiting histone variants or chromatin remoders promoting long-lasting chromatin relaxation. Importantly, strategies to neutralize the overexpression of DUX4-FL in mice have shown to be safe and provide benefits. To explore the role of DUX4-FL in the context of HIV transcription, we propose to identify and quantify DUX4 isoforms expressed in relevant HIV productive or latent models. This will ensure the significance of our studies, since DUX4-FL is not expected to be expressed in blood CD4+T cells. We will define the mechanism by which DUX4-FL upregulates HIV transcription. We will study the direct DUX4-FL recruitment to the HIV promoter, its impact on epigenetic marks, and uncover cellular proteins complexed with DUX4-FL via a multipronged approach, including proteomics, genomics and cell-based approaches. The overarching goal of this study is to explicitly establish whether DUX4-FL, or complexed proteins, are important for HIV transcriptional regulation, with the longstanding objective of exploiting these factors for HIV functional cure approaches.

摘要 HIV根除策略旨在清除潜伏感染记忆库中残留的HIV转录 CD4 + T细胞，在接受有效抗逆转录病毒疗法（ART）治疗的艾滋病毒感染者（PLWH）中持续存在。 这些细胞中含有一种前病毒，在HIV启动子处，当宿主细胞被连续招募时， 转录延伸因子、病毒达特蛋白和宿主染色质重塑物（称为达特- 介导的转录）。在不同的HIV潜伏期模型中，长期阻断这种残留的HIV转录， 使用达特的抑制剂，导致HIV上的异染色质化和RNA聚合酶II的损失， 启动子，沿着防止病毒反弹。因此，我们的前提是转录和染色质重塑， 抑制剂沿着与达特抑制剂一起可用于封闭-锁定功能固化方法，目的在于 减少ART期间的残留病毒血症并限制病毒反弹。此外，获得新的有见地的知识 研究这些因子与HIV转录之间的联系将有助于将它们作为抗病毒靶点。 我们已经发现了一个直接的正相关关系之间的表达全长同种型的表达， 双同源框蛋白4（DUX4-FL）表达和HIV感染在原代血液CD4 + T细胞中的作用。DUX4-FL 通过促进残留和Tat介导的HIV转录，强烈激活HIV转录活性。 此外，HIV增强DUX4-FL转录，这表明HIV与DUX4-FL之间存在正转录串扰。 DUX4-FL是一种强转录因子，其主要在疾病表型中表达，如 面肩肱型肌营养不良症和癌症，但从未被描述为调节艾滋病毒。在疾病中， DUX4-FL通常调节进化或免疫适应性基因的表达，导致毒性和/或 逃避免疫系统。DUX4-FL也被描述为通过募集来打开压实的染色质。 组蛋白变体或染色质重塑剂促进持久的染色质松弛。重要的是， 中和DUX4-FL在小鼠中的过表达已被证明是安全的并提供益处。 为了探索DUX4-FL在HIV转录过程中的作用，我们提出鉴定和定量DUX4-FL， 在相关HIV生产或潜伏模型中表达的同种型。这将确保我们研究的重要性， 因为DUX4-FL预期不会在血液CD4 + T细胞中表达。我们将定义一种机制， DUX4-FL上调HIV转录。我们将研究DUX4-FL对HIV启动子的直接募集， 影响表观遗传标记，并通过多管齐下的方法揭示与DUX4-FL复合的细胞蛋白质。 方法，包括蛋白质组学，基因组学和基于细胞的方法。 本研究的首要目标是明确确定DUX4-FL或复合蛋白是否与细胞凋亡相关。 重要的艾滋病毒转录调控，长期的目标是利用这些因素的艾滋病毒 功能性治愈方法。