PRIMATE MODEL TOWARDS HIV ERADICATION STRATEGIES

灵长类动物模型实现艾滋病毒根除策略

• 批准号: 8357336
• 负责人: THOMAS W NORTH
• 金额: $ 14.36万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357336
• 关键词: Animal Model; Antiviral Agents; Biological Assay; CCR5 gene; CD4 Positive T Lymphocytes; California; Clinical; DNA; Detection; Drug Combinations; Funding; Goals; Grant; HIV; HIV-1; Highly Active Antiretroviral Therapy; Human; Individual; Integrase Inhibitors; Latent Virus; Macaca; Macaca mulatta; Measures; Modeling; National Center for Research Resources; Pharmaceutical Preparations; Pharmacotherapy; Plasma; Population; Primates; Principal Investigator; Protease Inhibitor; RNA-Directed DNA Polymerase; Regimen; Research; Research Infrastructure; Residual state; Resources; Rest; Reverse Transcriptase Polymerase Chain Reaction; SIV; Source; Tissues; United States National Institutes of Health; Viral; Viral load measurement; Viremia; Virus; Virus Replication; antiretroviral therapy; cell type; cost; design; efavirenz; insight; non-nucleoside reverse transcriptase inhibitors; nucleoside analog; pinacolyl methylphosphonic acid; simian human immunodeficiency virus; viral DNA; viral RNA

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The long-term goal of this project is to develop and evaluate strategies for eradication of HIV-1 from infected individuals. These studies will be performed with an animal model that enables comprehensive analyses of viral reservoirs and replication dynamics during HAART. This model utilizes rhesus macaques infected by a chimeric virus of SIVmac239 containing the HIV-1 reverse transcriptase (RT) in place of the SIV RT (RT-SHIV). In this model a three-drug combination that is widely used in humans [efavirenz + (-)-FTC + PMPA] mimics HAART in HIV- 1-infected humans with respect to virus load (VL) suppression and rebound upon cessation of drug therapy. In order to study eradication strategies in the RT-SHIV/macaque model, we have developed a sensitive VL assay with a limit of detection of 1-2 copies of viral RNA (vRNA) per ml of plasma, and have also developed sensitive RT-PCR and PCR assays to measure vRNA and viral DNA (vDNA) in tissues. The goal of this project is to evaluate enhanced HAART in combinations that may more fully suppress plasma and tissue VLs to levels below detection with our sensitive PCR assays. Because a potential limitation of current antiretroviral therapy is access of drugs to key reservoir tissues, we will also determine levels of drugs in target tissues and use that information to optimize drug regimens. In addition to virological endpoints, the impact of these drug combinations will be evaluated in RT-SHIV-infected resting CD4+ lymphocyte populations. Importantly, the highly manipulatable RT-SHIV/macaque model enables assessment of potential clinical impact by determining effects on delay or reduction of viral rebound upon cessation of therapy. The drug combinations we propose are designed to evaluate: i) optimization of DNA chain termination with combinations of four potent nucleoside analogs (NRTI) in combination with efavirenz (NNRTI) or an integrase inhibitor, raltegravir; ii) combinations of the most effective NRTIs with both efavirenz and raltegravir; and iii) enhancement of the most potent drug combination with a protease inhibitor or a CCR5 antagonist. The overall Hypothesis of this project is that an intensified HAART regimen has the potential to eliminate RT-SHIV from infected macaques. These studies will provide insight on the requirements for complete suppression of virus replication in tissues and specific cell types as well as in plasma. Thus, this project aims to determine whether there is a limitation to suppression of residual viremia with antiviral drug combinations and whether eradication will also require "induction" strategies for reactivation of latent virus.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 该项目的长期目标是制定和评估从感染者身上根除 HIV-1 的策略。这些研究将使用动物模型进行，该模型能够在 HAART 期间全面分析病毒库和复制动态。该模型利用被 SIVmac239 嵌合病毒感染的恒河猴，该病毒含有 HIV-1 逆转录酶 (RT) 代替 SIV RT (RT-SHIV)。在该模型中，广泛用于人类的三药组合[依非韦伦 + (-)-FTC + PMPA] 在病毒载量 (VL) 抑制和停止药物治疗后反弹方面模拟 HIV-1 感染者的 HAART 。为了研究 RT-SHIV/猕猴模型中的根除策略，我们开发了一种灵敏的 VL 检测方法，检测限为每毫升血浆 1-2 个拷贝的病毒 RNA (vRNA)，并且还开发了灵敏的 RT-SHIV 检测方法。用于测量组织中 vRNA 和病毒 DNA (vDNA) 的 PCR 和 PCR 测定。该项目的目标是评估增强型 HAART 的组合，这些组合可以更充分地将血浆和组织 VL 抑制至低于我们灵敏的 PCR 检测检测到的水平。由于当前抗逆转录病毒治疗的一个潜在限制是药物进入关键储存组织，因此我们还将确定靶组织中的药物水平，并利用该信息来优化药物治疗方案。除了病毒学终点之外，还将在感染 RT-SHIV 的静息 CD4+ 淋巴细胞群中评估这些药物组合的影响。重要的是，高度可操作的 RT-SHIV/猕猴模型可以通过确定停止治疗后延迟或减少病毒反弹的效果来评估潜在的临床影响。我们提出的药物组合旨在评估： i) 四种有效核苷类似物 (NRTI) 与依非韦伦 (NNRTI) 或整合酶抑制剂拉替拉韦 (raltegravir) 组合的 DNA 链终止优化； ii) 最有效的 NRTI 与依非韦伦和拉替拉韦的组合； iii) 增强与蛋白酶抑制剂或 CCR5 拮抗剂的最有效药物组合。该项目的总体假设是强化HAART疗法有可能消除受感染猕猴体内的RT-SHIV。这些研究将深入了解完全抑制病毒在组织和特定细胞类型以及血浆中复制的要求。因此，该项目旨在确定抗病毒药物组合对抑制残留病毒血症是否存在限制，以及根除病毒是否还需要“诱导”策略来重新激活潜伏病毒。