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：
8930058
负责人：
OLAF KUTSCH
金额：
$ 33.08万
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：
2014
资助国家：
美国
起止时间：
2014-09-19 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8930058
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 Goals HIV HIV Infections HIV-1 Hand Health 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 Proviruses Reporting Rest Sampling Signal Pathway Signal Transduction Sorting - Cell Movement Stimulus T memory cell T-Lymphocyte Technology Testing Therapeutic Therapeutic Intervention Time Viral Virus anergy base bryostatin clinically relevant cytokine drug candidate drug discovery drug efficacy improved in vivo insight latent infection new technology novel prostratin protein protein interaction reconstitution response success transcription factor

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 Goals HIV HIV Infections HIV-1 Hand Health 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 Proviruses Reporting Rest Sampling Signal Pathway Signal Transduction Sorting - Cell Movement Stimulus T memory cell T-Lymphocyte Technology Testing Therapeutic Therapeutic Intervention Time Viral Virus anergy base bryostatin clinically relevant cytokine drug candidate drug discovery drug efficacy improved in vivo insight latent infection new technology novel prostratin protein protein interaction reconstitution response success transcription factor

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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：潜在的HIV-1感染已被认为是治愈HIV-1治疗发展的主要障碍，但是，仍定义了控制潜在感染的细胞和病毒相互作用事件。在过去几年中，已促进的HIV重新激活的HIV重新激活的HDAC抑制剂的使用引起了人们的意识，应检查艾滋病毒可以并且应该检查。然而，HDAC抑制剂作为HIV-1重新激活的剂的成功受到限制。该应用的关键是认识到潜在的HIV-1感染事件的宿主细胞实际上以迫使病毒进入A的方式进行表型改变潜在状态并使细胞对刺激无反应。尽管连续暴露于同源抗原，但宿主T细胞的反应状，无反应的状态将最终解释LATNT病毒晶状体的体内稳定性。这也将解释最近的发现，描述了大多数在体内患者材料中的潜在感染事件甚至对PHA刺激无反应。我们先前证明了即使在高水平的诱导NF-KB活性的情况下，也存在控制潜在HIV-1感染的网守酶激酶功能。现在，我们的数据表明，宿主细胞的表型变化超出了单个激酶的功能，但是在潜在感染的T细胞中，所有途径的活性都会改变。 Kinome分析揭示了我们已经成功靶向的新型药物靶标，以改变T细胞系中潜在HIV-1感染的控制和潜在HIV感染的原发性T CLL模型。我们还提供了数据，表明来自HIV-1患者的CD4 T细胞成功地在激化水平上大大改变了。虽然与T细胞系中的激活变化更为复杂，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细胞系中获得的某些发现可以直接传递到主要T细胞模型中，并用于在该应用程序中使用潜在的HIV的稳定性，并在此应用程序中为此提供了细胞，以便在此应用程序中获得群体的详细信息。使用来自HIV-1感染患者的离体T细胞材料的HIV-1感染事件，并产生描述HIV-1潜伏期控制的蛋白质蛋白相互作用网络（PIN）。然后，生成的引脚将用于探测控制潜在HIV-1感染的靶标，并开发有效触发HIV-1重新激活的新型药物组合。该应用的目的是建立一个潜在的HIV-1感染的综合模型，该模型考虑了病毒在激酶，转录因子和可能的染色质水平上与宿主细胞与宿主细胞的动态，双向相互作用，并利用这种知识来驱动药物重新定位的努力，以识别该药物的组合，从而逆转其在此的宿主T细胞，从而逆转宿主T细胞的状态。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意外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。