Targeting regulators of cellular gene transcription to impact HIV latency

靶向细胞基因转录调节因子以影响 HIV 潜伏期

• 批准号: 7554818
• 负责人: DOUGLAS D RICHMAN
• 金额: $ 90.08万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7554818
• 关键词: Targeting; regulators; cellular; gene; transcription

DESCRIPTION (provided by applicant): The impact of antiretroviral therapy represents a major demonstration of the capability and success of modern drug discovery, development, and implementation. Nevertheless, in infected individuals this success lasts only as long as adherence to the regimen is maintained. Discontinuation of treatment results in viral rebound, necessitating the chronic, lifelong administration of antiretroviral therapy. The requirement for lifelong therapy results from the inability to deplete the persistent reservoir of latently infected cells. The admittedly ambitious ultimate goal of this application is to purge the latently infected reservoir with the objective of eradicating the infection or minimizing the HIV reservoir sufficiently to attain durable immunologic control. This Program Project has been designed with three Projects and two Scientific Cores. Project 1 will identify and characterize potent histone deacetylase inhibitors for the initial studies, while using a human genome siRNA library to identify additional targets for small molecule intervention. These treatment strategies will be systematically evaluated in the following models: (1) the in vitro model of latently infected primary human CD4 lymphocytes developed in our laboratory (Project 2), (2) CD4 lymphocytes obtained ex vivo from chronically infected, HAART-suppressed patients (project 2), and (3) an SIV/Rhesus macaque model (Project 1). Latent infection will be quantified by the two scientific cores. The Molecular Virology Core will measure infectivity, HIV (SIV) nucleic acid species including integrated DNA, and ability of CD4 lymphocytes to express p24 (p27) upon activation. The Molecular Imaging Core (Haase lab) will quantify and characterize infection in antiretroviral drug-suppressed cells from the in vitro HIV model and from macaque tissues. While assessing treatment efficacy and minimizing toxicity, potential collateral immunological consequences of these interventions targeted to host functions will be systematically characterized in Project 3. This highly coordinated, collaboration which ranges from target and drug discovery through animal model proof-of-concept will reveal new insights regarding the pathogenetic mechanisms of HIV latency; however, the goal of the Program Project is to cure AIDS, PROJECT 1: (Hazuda, D) PROJECT 1 DESCRIPTION (provided by applicant): The long term goal of therapy for HIV-1 infection should be to develop treatment regimens either to provide durable control of viral replication and/or eradicate the infection. Among the limitations of current therapy are the incomplete suppression of viral replication in many patients and the inability to affect the latent population of HIV-1 in quiescent CD4+ T-lymphocytes. While new agents in development may enhance the potency and durability of antiretroviral treatments, these will not address the longer term problem of latency. The current program application proposes to evaluate the hypothesis that histone deacetylase (HDAC) inhibitors or other LTR-activating compounds can affect latent infection in a rhesus macaque model of retroviral latency. This hypothesis is based on in vitro observations suggesting that HIV-1 latency is maintained by cellular mechanisms that control chromatin structure and that derepression of HIV-1 transcription can be induced by HDAC inhibitors without activating T-cells. In Project 1 of the application, we will identify and provide inhibitors as well as carry out a genome-scale siRNA screen to identify LTR-suppressing factors to identify different pathways for derepressing HIV-1 transcription. Additionally, we will develop critical assays required to establish an SIV model to test this hypothesis and support the evaluation of the HDAC inhibitor, vorinostat, and other LTR activators in vivo. The objectives of Project 1 include providing potent integrase inhibitors for a novel combination regimen (ART) that will be effective in SIV infection, providing vorinostat and other LTR-activating compounds and evaluating their effects on SIV latency, establishing pharmacodynamic assays for HDAC activity that will be used to select doses and identifying novel LTR-activators with enhanced potency and selectivity for testing in this model. The objective is to assess the susceptibility of the latent reservoirs to therapeutic intervention with the ultimate goal of identifying inhibitors and establishing treatment paradigms suitable for clinical testing.

描述(申请人提供)：抗逆转录病毒治疗的影响代表了现代药物发现、开发和实施的能力和成功的主要证明。然而，在受感染的人中，这种成功只持续到坚持治疗方案的情况下。停止治疗会导致病毒反弹，需要长期、终身地进行抗逆转录病毒治疗。需要终生治疗的原因是无法耗尽潜伏感染细胞的持久储存库。诚然，这一应用程序的最终目标是清除潜伏感染的宿主，目的是根除感染或将艾滋病毒宿主降至最低，以获得持久的免疫控制。该计划项目由三个项目和两个科学核心组成。项目1将在最初的研究中识别和表征有效的组蛋白去乙酰化酶抑制剂，同时使用人类基因组siRNA文库来识别小分子干预的其他靶点。这些治疗策略将在以下模型中进行系统评估：(1)我们实验室开发的潜伏感染原代人类CD4淋巴细胞的体外模型(项目2)，(2)从慢性感染、HAART抑制的患者体内获得的CD4淋巴细胞(项目2)，以及(3)SIV/恒河猴模型(项目1)。潜伏感染将通过这两个科学核心进行量化。分子病毒学核心将测量传染性、艾滋病毒(SIV)核酸种类(包括整合的DNA)以及CD4淋巴细胞在激活时表达p24(P27)的能力。分子成像核心(HAASE LAB)将量化和表征体外艾滋病毒模型和猕猴组织中抗逆转录病毒药物抑制细胞的感染情况。在评估治疗效果和将毒性降至最低的同时，这些针对宿主功能的干预措施的潜在附带免疫后果将在项目3中得到系统表征。这种从靶向和药物发现到动物模型概念验证的高度协调的合作将揭示关于艾滋病毒潜伏期的致病机制的新见解；然而，该项目的目标是治愈艾滋病， 项目1：(哈祖达，D) 项目1说明(申请人提供)：艾滋病毒-1感染治疗的长期目标应该是开发治疗方案，以提供对病毒复制的持久控制和/或根除感染。目前治疗的局限性包括对许多患者病毒复制的不完全抑制，以及无法影响静止的CD4+T淋巴细胞中潜伏的HIV-1群体。虽然正在开发的新药物可能会增强抗逆转录病毒治疗的效力和持久性，但这些不会解决潜伏期的长期问题。目前的程序应用程序建议评估组蛋白脱乙酰酶(HDAC)抑制剂或其他LTR激活化合物可以影响逆转录病毒潜伏期猕猴模型中的潜伏感染的假设。这一假说是基于体外观察，表明HIV-1潜伏期由控制染色质结构的细胞机制维持，HDAC抑制剂可以在不激活T细胞的情况下诱导HIV-1转录的去抑制。在应用程序的项目1中，我们将识别和提供抑制剂，并进行基因组规模的siRNA筛选，以确定LTR抑制因子，以确定不同的途径来降低HIV-1的转录。此外，我们将开发建立SIV模型所需的关键分析来验证这一假说，并支持体内对HDAC抑制剂、涡旋器和其他LTR激活剂的评估。项目1的目标包括为将在SIV感染中有效的新型联合疗法(ART)提供有效的整合酶抑制剂，提供涡旋剂和其他LTR激活化合物并评估它们对SIV潜伏期的影响，建立将用于选择剂量的HDAC活性的药效学分析，并确定具有增强效力和选择性的新型LTR激活剂用于该模型的测试。其目的是评估潜在储存库对治疗干预的敏感性，最终目的是确定抑制剂并建立适合临床试验的治疗范例。