Developing Pathogen Recognition Receptor Agonists as Latency Reversing Agents

开发病原体识别受体激动剂作为潜伏期逆转剂

基本信息

批准号：
9501675
负责人：
Alberto Bosque
金额：
$ 49.61万
依托单位：
GEORGE WASHINGTON UNIVERSITY
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：
2016-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9501675
关键词：
Adjuvant Agonist Anti-Retroviral Agents Antiviral Agents Asthma Autoimmunity Bacterial Infections Bromodomain CD4 Positive T Lymphocytes Cell model Cells Clinical Clinical Trials Communicable Diseases Consequentialism Data Disease Disease remission Epigenetic Process Evaluation Gene Expression Genetic Transcription Goals Guanosine Triphosphate Phosphohydrolases HIV HIV-1 HIV-1 protease Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor In Vitro Infection Interferons Interphase Cell Investigation Latent Virus Lead Ligands MGMT gene Maintenance Mitochondria Mitochondrial Proteins Modeling Nucleic Acids Pathway interactions Patients Pharmaceutical Preparations Pharmacology Protein Kinase C Proteins Role SIRT1 gene Shock Signal Transduction Signaling Protein T-Cell Activation Technology Therapeutic Thinking Tumor Cell Line Vaccination Viral Viral Genes Viral reservoir Virus Diseases Virus Latency base bryostatin cancer therapy design immune activation inhibitor/antagonist latent infection novel novel strategies novel therapeutic intervention pathogen reactivation from latency receptor response screening synergism therapeutic target

Adjuvant Agonist Anti-Retroviral Agents Antiviral Agents Asthma Autoimmunity Bacterial Infections Bromodomain CD4 Positive T Lymphocytes Cell model Cells Clinical Clinical Trials Communicable Diseases Consequentialism Data Disease Disease remission Epigenetic Process Evaluation Gene Expression Genetic Transcription Goals Guanosine Triphosphate Phosphohydrolases HIV HIV-1 HIV-1 protease Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor In Vitro Infection Interferons Interphase Cell Investigation Latent Virus Lead Ligands MGMT gene Maintenance Mitochondria Mitochondrial Proteins Modeling Nucleic Acids Pathway interactions Patients Pharmaceutical Preparations Pharmacology Protein Kinase C Proteins Role SIRT1 gene Shock Signal Transduction Signaling Protein T-Cell Activation Technology Therapeutic Thinking Tumor Cell Line Vaccination Viral Viral Genes Viral reservoir Virus Diseases Virus Latency base bryostatin cancer therapy design immune activation inhibitor/antagonist latent infection novel novel strategies novel therapeutic intervention pathogen reactivation from latency receptor response screening synergism therapeutic target

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项目摘要

The existence of latent reservoirs of HIV-infected cells constitutes the major impediment towards viral eradication. HIV-1 latent reservoirs are small, but extremely long-lived. Latent infection is associated with undetectable levels of viral gene expression and appears to be non-cytopathic. However, upon reactivation, latent viruses enter an active mode of replication in which they are fully competent for spread and induction of disease. The current thinking in the field is that a combination of hypothetical drugs that will reactivate latent viruses (Latency Reversing Agent or LRA), with present-day antiretroviral drugs, will be an effective approach toward viral eradication. We have previously found that Pam3CSK4, a TLR-1/2 agonist, can reactivate latent HIV-1. Based on this earlier finding, we propose to further investigate the possibility of developing Pathogen Recognition Receptor (PRR) agonists as novel strategies to reactivate/eradicate latent HIV-1 infection. In Aim 1, we plan to characterize the ability of multi-PRR agonists to reactivate latent HIV-1. Multi-PRR agonists are synthetic TLR agonists developed by InvivoGen to be used as adjuvants in cancer therapy, infectious diseases and vaccination. Our preliminary data using the tumoral cell line JLat, a primary model of HIV-1 latency (Cultured TCM model) and cells isolated from aviremic patients demonstrate that these ligands have the ability to reactivate latent HIV-1. Furthermore, in this aim we also plan to evaluate the potential of this agonist to reduce the latent reservoir in vitro. We have found that dynasore, a GTPase inhibitor, has the ability to reactivate latent HIV-1. It has been proposed that dynasore activates the PRR mitochondria antiviral signal protein (MAVS). We have further corroborated this result. Based on this notion, this aim is designed to characterize whether manipulation of the RIG-I/MDA-5/MAVS pathway can have therapeutic implications towards HIV-1 eradication. First, we are going to study whether RIG-1 and MDA-5 ligands that signal through MAVS can reactivate latent HIV-1. Second, it has been shown that the HIV-1 protease can induce lysosomal degradation of RIG-I to avoid sensing and the induction of an interferon response. We plan to characterize whether this mechanism is involved in the establishment of latency. Lastly, identifying novel mechanisms that are involved in the maintenance of latency would lead to the design of novel therapeutic approaches. We have performed a screening to identify epigenetic modulators that synergize with the synthetic TLR-2 ligand CL572. We have identified several pathways that synergize with CL572. In Aim 3, we plan to characterize in detail the mechanism of synergy between PRR agonists and two known pathways, HDACs and bromodomain proteins, and two novel pathways, O6-Methylguanine-DNA methyltransferase and sirtuin-1 and -2. The overall goal of this proposal is to move forward novel targets that can be exploited therapeutically to either eradicate viral reservoirs for HIV-1 infections or to induce a long-term remission without treatment (functional cure).

HIV感染细胞的潜在储层的存在构成了病毒的主要障碍根除。 HIV-1潜在水库很小，但寿命非常长。潜在感染与无法检测到的病毒基因表达水平，并且似乎是非环保性的。但是，重新激活后，潜在病毒进入一种主动复制方式，在该模式中，它们完全有权传播和诱导疾病。现场的当前思想是，假设药物将重新激活潜在药物病毒（潜伏期逆转剂或LRA），使用当今的抗逆转录病毒药物，将是一种有效的方法朝着消除病毒。我们以前已经发现，TLR-1/2激动剂PAM3CSK4可以重新激活潜在 HIV-1。基于这一早期发现，我们建议进一步研究发展病原体的可能性识别受体（PRR）激动剂是重新激活/根除潜在HIV-1感染的新策略。在AIM 1中，我们计划表征多PRR激动剂重新激活潜在HIV-1的能力。多prr 激动剂是由Invivogen开发的合成TLR激动剂，用作癌症治疗中的佐剂，传染病和疫苗接种。我们使用肿瘤细胞系JLAT的初步数据，这是一个主要模型 HIV-1潜伏期（培养的TCM模型）和从无毒患者分离的细胞表明这些配体具有重新激活潜在HIV-1的能力。此外，在此目标中，我们还计划评估这一潜力激动剂在体外减少潜在储层。我们发现GTPase抑制剂Dynasore具有能力重新激活潜在的HIV-1。已经提出Dynasore激活PRR线粒体抗病毒信号蛋白质（MAV）。我们进一步证实了这一结果。基于这个概念，此目标旨在表征对RIG-I/MDA-5/MAVS途径的操纵是否具有治疗意义朝着消除HIV-1。首先，我们将研究rig-1和MDA-5通过信号的配体 MAV可以重新激活潜在的HIV-1。其次，已经表明HIV-1蛋白酶可以诱导溶酶体 RIG-1的降解以避免感应和诱导干扰素反应。我们计划表征这种机制是否参与了延迟的建立。最后，确定新的机制参与延迟的维持将导致新型治疗方法的设计。我们有进行了筛选，以识别与合成TLR-2配体CL572协同作用的表观遗传调节剂。我们已经确定了与CL572协同作用的几种途径。在AIM 3中，我们计划详细描述 PRR激动剂与两个已知途径HDACS和溴化域蛋白之间的协同作用机理，以及两种新型途径，O6-甲基鸟嘌呤-DNA甲基转移酶和Sirtuin-1和-2。该提案的总体目标是推进可以通过治疗方法利用的新颖目标要么消除病毒储存库HIV-1感染，要么在未经治疗的情况下诱导长期缓解（功能治疗）。