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（由申请人提供）：抗逆转录病毒疗法的影响代表了现代药物发现、开发和实施的能力和成功的重要证明。然而，对于感染者来说，只有坚持坚持治疗方案，这种成功才能持续。停止治疗会导致病毒反弹，需要长期、终生服用抗逆转录病毒治疗。终生治疗的要求是由于无法耗尽潜伏感染细胞的持久储存库。诚然，该应用雄心勃勃的最终目标是清除潜伏感染的病毒库，以消除感染或充分减少艾滋病毒病毒库，以获得持久的免疫控制。该计划项目设计有三个项目和两个科学核心。项目 1 将鉴定和表征用于初步研究的有效组蛋白脱乙酰酶抑制剂，同时使用人类基因组 siRNA 库来鉴定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CD4淋巴细胞的体外模型（项目2），（2）从慢性感染、HAART抑制的体外获得的CD4淋巴细胞患者（项目 2），以及（3）SIV/恒河猴模型（项目 1）。潜伏感染将通过两个科学核心进行量化。分子病毒学核心将测量感染性、HIV (SIV) 核酸种类（包括整合 DNA）以及 CD4 淋巴细胞激活后表达 p24 (p27) 的能力。分子成像核心（Haase 实验室）将量化和表征来自体外 HIV 模型和猕猴组织的抗逆转录病毒药物抑制细胞的感染。在评估治疗效果和最大限度地减少毒性的同时，项目 3 将系统地描述这些针对宿主功能的干预措施的潜在附带免疫后果。这种高度协调的合作范围从靶点和药物发现到动物模型概念验证，将揭示新的关于艾滋病毒潜伏期发病机制的见解；然而，该计划项目的目标是治愈艾滋病， 项目 1：（Hazuda，D） 项目 1 描述（由申请人提供）：HIV-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许多患者体内病毒复制的抑制不完全，以及无法影响静态 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，但这些并不能解决长期的潜伏期问题。当前的计划申请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恒河猴模型中的潜伏感染的假设。这一假设基于体外观察，表明 HIV-1 潜伏期是通过控制染色质结构的细胞机制维持的，并且 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。在该申请的项目1中，我们将鉴定并提供抑制剂，并进行基因组规模的siRNA筛选，以鉴定LTR抑制因子，从而确定去抑制HIV-1转录的不同途径。此外，我们将开发建立 SIV 模型所需的关键测定方法，以测试这一假设并支持 HDAC 抑制剂、伏立诺他和其他 LTR 激活剂的体内评估。项目 1 的目标包括为有效治疗 SIV 感染的新型联合治疗方案 (ART) 提供有效的整合酶抑制剂，提供伏立诺他和其他 LTR 激活化合物并评估它们对 SIV 潜伏期的影响，建立 HDAC 活性的药效学测定，用于选择剂量并鉴定具有增强效力和选择性的新型 LTR 激活剂，以用于该模型中的测试。目的是评估潜在储库对治疗干预的敏感性，最终目标是识别抑制剂并建立适合临床测试的治疗范例。