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）：抗逆转录病毒治疗的影响是现代药物发现、开发和实施的能力和成功的主要证明。然而，在受感染的个体中，这种成功仅持续到坚持该方案。停止治疗会导致病毒反弹，需要长期、终身给予抗逆转录病毒治疗。终身治疗的要求是由于无法耗尽潜伏感染细胞的持久储存库。该应用的公认的雄心勃勃的最终目标是清除潜伏感染的储库，目的是根除感染或使HIV储库充分最小化以获得持久的免疫控制。该计划项目设计了三个项目和两个科学核心。项目1将为初步研究鉴定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文库鉴定小分子干预的其他靶点。将在以下模型中系统评价这些治疗策略：（1）我们实验室开发的潜伏感染原代人CD 4淋巴细胞体外模型（项目2），（2）从慢性感染、HAART抑制患者中离体获得的CD 4淋巴细胞（项目2），和（3）SIV/恒河猴模型（项目1）。潜在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（包括整合的DNA）和CD 4淋巴细胞在活化后表达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中的感染。在评估治疗疗效和最大限度地减少毒性的同时，这些针对宿主功能的干预措施的潜在附带免疫学后果将在项目3中进行系统表征。这种高度协调的合作，从目标和药物发现到动物模型概念验证，将揭示有关艾滋病毒潜伏期发病机制的新见解;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是开发治疗方案，以持久控制病毒复制和/或根除感染。当前疗法的局限性包括对许多患者的病毒复制抑制不完全，以及无法影响静止CD 4 + T淋巴细胞中HIV-1的潜伏群体。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，但这些药物无法解决潜伏期的长期问题。目前的计划申请提出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的恒河猴模型中的潜伏感染的假设。这一假设是基于体外观察，表明HIV-1潜伏期是由控制染色质结构的细胞机制维持的，HDAC抑制剂可以诱导HIV-1转录的去抑制而不激活T细胞。在申请的项目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，从而鉴定去抑制HIV-1转录的不同途径。此外，我们将开发建立SIV模型所需的关键试验，以检验这一假设，并支持HDAC抑制剂、伏立诺他和其他LTR激活剂的体内评价。项目1的目的包括提供有效的整合酶抑制剂用于新的组合方案（ART），其将在SIV感染中有效，提供伏立诺他和其他LTR活化化合物并评估其对SIV潜伏期的影响，建立HDAC活性的药效学测定，其将用于选择剂量，并鉴定具有增强的效力和选择性的新型LTR活化剂用于在该模型中进行测试。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试验的治疗模式。