Kinomic analysis of host cell factors controlling latent HIV-1 infection

控制潜伏 HIV-1 感染的宿主细胞因子的基因组分析

基本信息

批准号：
9325418
负责人：
OLAF KUTSCH
金额：
$ 44.1万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9325418
关键词：
AIDS therapy Affect Antigens Awareness CD28 gene CD3 Antigens CD4 Positive T Lymphocytes Cell Line Cell model Cells Chromatin Clinical Complex Data Development Drug Combinations Drug Compounding Drug Targeting Event Exposure to FDA approved Future Gatekeeping Goals HIV HIV Infections HIV-1 Hand Histone Deacetylase Inhibitor Individual Infection Knowledge Measurable Measures Modeling Molecular Molecular Biology Molecular Target NF-kappa B Pathway interactions Patients Pharmaceutical Preparations Phase Phenotype Phosphotransferases Population Process Proteins Proviruses Reporting Rest Sampling Signal Pathway Stimulus T memory cell T-Lymphocyte Technology Testing Therapeutic Therapeutic Intervention Viral Viral reservoir Virus Virus Latency anergy base bryostatin clinically relevant cytokine drug candidate drug discovery drug efficacy drug testing improved in vivo insight latent infection molecular drug target new technology new therapeutic target novel novel drug combination prostratin protein protein interaction public health relevance reconstitution response success transcription factor

项目摘要

DESCRIPTION: Latent HIV-1 infection has been recognized as one major obstacle to the development of a curative HIV-1 therapy, however, cellular and viral interaction events that control latent infection are still ill defined. The use of HDAC inhibitors that have been promoted as HIV reactivating drugs in the last years has generated awareness that HIV eradication can and should be examined. However, the success of HDAC inhibitors as HIV-1 reactivating agents has been limited. Key to this application is the realization that the host-cells of latent HIV-1 infection events are actually phenotypically altered in a manner that forces the virus into a latent state and renders the cells unresponsive to stimulation. An anergy-like, unresponsive state of the host T cells would conclusively explain the extraordinary in vivo stability of the latnt viral reservoir despite continuous exposure to cognate antigen. It would also explain recent findings describing that the majority of the latent infection events in ex vivo patient material ar even unresponsive to PHA stimulation. We previously demonstrated the presence of a gate-keeper kinase function that controls latent HIV-1 infection even in the presence of high levels of induced NF-kB activity. Our data now show that phenotypic changes of the host cells go beyond the functionality of individual kinases, but that the activities of entire pathways are altered in latently infected T cells. Kinome profiling revealed novel drug targets that we have already successfully targeted to alter control of latent HIV-1 infection in T cell lines and in primary T cll models of latent HIV infection. We also provide data that CD4 T cells from HIV-1 patients on successful ART are massively altered at the kinomic level. While the kinomic changes are more complex than in T cell lines, there is complete overlap in the affected T cell signaling pathways, a finding that likely explains why some of the findings obtained in T cell lines can be directly transferred into primary T cell models and used to perturb the stability of the latent HIV reservoir. Given this observation, in this application, we propose to generate increasingly detailed kinome profiles of the most relevant memory T cell sub-populations know to host latent HIV-1 infection events using ex vivo T cell material from HIV-1 infected patients and to generate protein-protein interaction networks (PIN) that describe HIV-1 latency control. The generated PINs will then be used to probe identified targets that control latent HIV- 1 infection and to develop novel drug combinations that efficiently trigger HIV-1 reactivation. The goal of this application is to establish a comprehensive model of latent HIV-1 infection that considers the dynamic, bi-directional interactions of the virus with the host-cell at the kinase, transcription factor and possibly chromatin level and to use this knowledge to drive a drug repositioning effort to identify drug combinations that will reverse the unresponsive state of the host T cells, thereby allowing cognate antigen and possible therapeutic stimuli to trigger HIV-1 reactivation. The insights gained from the described studies will provide a blueprint for how to develop efficient HIV-1 reactivating therapeutic strategies based on unexplored molecular drug targets.

描述：潜伏性 HIV-1 感染已被认为是开发治疗性 HIV-1 疗法的主要障碍，然而，控制潜伏性感染的细胞和病毒相互作用事件仍不清楚。已推广的 HDAC 抑制剂的使用。近年来，HIV 再激活药物已经引起了人们的认识，即可以而且应该对 HIV 根除进行研究。然而，HDAC 抑制剂作为 HIV-1 再激活药物的成功程度有限，这一应用的关键是认识到宿主细胞。潜伏的 HIV-1 感染事件的表型实际上发生了改变，迫使病毒进入潜伏状态并使细胞对刺激无反应，这将最终解释潜伏病毒库在连续暴露于同源抗原的情况下仍具有非凡的体内稳定性。离体患者材料中的大多数潜伏感染事件甚至对 PHA 刺激没有反应，我们之前证明了控制潜伏 HIV-1 感染的看门人激酶功能的存在。即使存在高水平的诱导 NF-kB 活性，我们的数据现在表明宿主细胞的表型变化超出了单个激酶的功能，但在潜伏感染的 T 细胞中整个通路的活性发生了改变。揭示了新的药物靶标，我们已经成功地针对这些靶标来改变 T 细胞系和潜伏 HIV 感染的原代 T cll 模型中对潜伏 HIV-1 感染的控制。我们还提供了来自 HIV-1 患者的 CD4 T 细胞成功接受 ART 的数据。是虽然动组学变化比 T 细胞系更复杂，但受影响的 T 细胞信号传导途径完全重叠，这一发现可能解释了为什么在 T 细胞系中获得的一些发现可以直接进行。转移到原代 T 细胞模型中并用于扰乱潜伏 HIV 储存库的稳定性鉴于这一观察，在本应用中，我们建议生成已知宿主潜伏 HIV 的最相关记忆 T 细胞亚群的越来越详细的激酶组谱。 1 使用离体感染事件来自 HIV-1 感染患者的 T 细胞材料，并生成描述 HIV-1 潜伏控制的蛋白质-蛋白质相互作用网络 (PIN)，然后使用生成的 PIN 来探测控制潜伏 HIV-1 感染的已识别靶点并开发新的靶点。该应用的目标是建立一个潜在的 HIV-1 感染的综合模型，该模型考虑了病毒与宿主细胞在激酶、转录因子处的动态、双向相互作用。以及可能的染色质水平利用这些知识推动药物重新定位工作，以确定能够逆转宿主 T 细胞无反应状态的药物组合，从而允许同源抗原和可能的治疗刺激触发 HIV-1 重新激活从所描述的研究中获得的见解将为如何基于未探索的分子药物靶标开发有效的 HIV-1 重新激活治疗策略提供蓝图。