Targeting the host kinase DYRK1A to optimize reversal of HIV-1 latency in CD4 T cells

靶向宿主激酶 DYRK1A 以优化 CD4 T 细胞中 HIV-1 潜伏期的逆转

• 批准号: 9312939
• 负责人: Mark A Wallet
• 金额: $ 37.56万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9312939
• 关键词: AIDS/HIV problem; Address; Affinity; Antigens; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Line; Cell physiology; Cells; Cessation of life; Chemicals; Combined Modality Therapy; Cytotoxic T-Lymphocytes; Data; Development; Effector Cell; Failure; Frequencies; Future; Gene Expression; Genes; Goals; HIV; HIV Genome; HIV Infections; HIV-1; Harmine; Health; Human; Individual; Infection; Integration Host Factors; Investigation; Knowledge; Laboratories; Lead; Mediating; Mission; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Process; Proteins; Proviruses; Public Health; Research; Role; Shock; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Specificity; Structure; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Toxic effect; Tyrosine Phosphorylation; United States National Institutes of Health; Viral Cytopathogenic Effect; Virus; Virus Replication; Work; antiretroviral therapy; base; design; genetic regulatory protein; improved; inhibitor/antagonist; innovation; killings; knock-down; latent persistent infection; new therapeutic target; novel; novel strategies; novel therapeutics; nuclear factors of activated T-cells; prevent; promoter; purge; response; small hairpin RNA; transcription factor

 DESCRIPTION (provided by applicant): There is a significant unmet need to develop therapeutics that could cure HIV infection by eliminating reservoir cells where the virus resides during antiretroviral therapy [ART]. This need persists because of gaps in knowledge of how HIV maintains persistent latent infection of reservoir cells. These gaps represent an important problem because until they are filled a cure for HIV remains impossible. The long term goal is to more completely understand host mechanisms that regulate HIV latency versus active replication. The immediate objective of this application is to prove that pharmacologic inhibitors of dual specificity tyrosine- phosphorylation-regulated kinase 1A [DYRK1A] allow optimal reactivation of latent HIV from CD4 T cells. To achieve this objective, a combination of latently HIV-infected cell lines and primary CD4 T cells will be utilized to investigate the role of DYRK1A in maintaining HIV latency. The central hypothesis is that DYRK1A facilitates HIV latency by negatively regulating host transcription factors that drive the HIV LTR promoter. This hypothesis is based upon preliminary studies performed in the applicant's laboratory demonstrating that the DYRK1A inhibitor harmine boosts the efficacy of existing latency reversing agents [LRAs] and enhances LRA-induced NFAT activity. Furthermore, knockdown of DYRK1A mimics the effects of harmine. A secondary hypothesis is that DYRK1A inhibits antigen-induced activation of CD8 T cells, thus contributing to the failure of patient CD8 T cells to kill HIV reservoir cells wherei HIV replication has been triggered by LRAs. We demonstrate that harmine enhances antigen-induced CD8 T cell activation. Thus, our pharmacologic approach is entirely novel because it simultaneously improves reactivation of latent HIV and improves activity of the CD8 effector cells needed to kill off the HIV reservoir. The rationale for the proposed study is that understanding host mechanisms that regulate HIV latency and replication will lead to development of new drugs to maximize reactivation of latent HIV and to maximize death of reservoir cells that carry HIV genomes. Guided by preliminary studies, the hypothesis will be objectively tested by pursuing three specific aims: 1) Establish the efficacy of DYRK1A inhibitors as anti-latency drugs; 2) Identify host transcription factors that are required for optimal HIV reactivation from latently-infected CD4 T cells; and 3) Establish the efficacy of DYRK1A inhibitors for enhancing death of reactivated HIV reservoir cells. The approach is innovative because it uses inhibitors of a specific host regulatory protein, DYRK1A, to better understand mechanisms that control HIV gene expression and viability of reservoir cells as well as CD8 T cell activation. The proposed research is significant because it is expected to advance a new class of anti-latency drugs (DYRK1A inhibitors) and to increase mechanistic understanding of HIV latency as a basis for new research based on expected outcomes. Ultimately, the knowledge gained from this work has the potential to support a combination therapeutic cure for HIV infection.

 描述（由适用提供）：通过消除抗逆转录病毒治疗期间病毒居住的储层细胞来培养可以治愈HIV感染的理论的重要需求[ART]。由于知识如何保持储层细胞的持续潜在感染，因此这种需求持续存在。这些差距代表了一个重要的问题，因为直到填补艾滋病毒的治疗方法之前，仍然是不可能的。长期目标是更完全了解调节艾滋病毒潜伏期与主动复制的宿主机制。该应用的直接目的是证明双重特异性酪氨酸 - 磷酸化调节的激酶1A [DYRK1A]的药物抑制剂允许从CD4 T细胞中最佳地重新激活潜在的HIV。为了实现这一目标，将利用潜在的HIV感染细胞系和原代CD4 T细胞的组合来研究DYRK1A在维持HIV潜伏期中的作用。中心假设是DYRK1A通过负控制驱动HIV LTR启动子的宿主转录因子来宿主艾滋病毒潜伏期。该假设基于在应用程序实验室中进行的初步研究，证明DYRK1A抑制剂Harmine提高了现有潜伏期逆转剂的效率[LRAS]并增强了LRA诱导的NFAT活性。此外，DYRK1A的敲低模仿Harmine的影响。次要假设是DYRK1A抑制了抗原诱导的CD8 T细胞激活，因此LRA触发了患者CD8 T细胞杀死HIV复制的失败。我们证明Harmine增强了抗原诱导的CD8 T细胞活化。这就是我们的药物方法完全新颖，因为它只是改善了潜在艾滋病毒的重新激活并改善了杀死HIV储量所需的CD8效应细胞的活性。拟议的研究的理由是，了解调节HIV潜伏期和复制的宿主机制将导致新药的发展，以最大程度地激活潜在的HIV并最大化携带HIV基因组的储层细胞死亡。在初步研究的指导下，该假设将通过追求三个具体目的来客观检验：1）确定DYRK1A抑制剂作为抗延迟药物的有效性； 2）确定来自潜在感染的CD4 T细胞最佳HIV重新激活所需的宿主转录因子； 3）建立DYRK1A抑制剂增强重新激活HIV储层细胞死亡的有效性。该方法具有创新性，因为它使用了特定宿主调节蛋白DyRK1A的抑制剂，以更好地理解控制HIV基因表达的机制和RESERVER细胞以及CD8 T细胞激活的机制。拟议的研究之所以重要，是因为预计它将推进一类新的抗延迟药物（DYRK1A抑制剂），并提高对HIV潜伏期的机械理解，作为基于预期结果的新研究的基础。最终，从这项工作中获得的知识有可能支持联合治疗艾滋病毒感染的方法。