Testing of VIF Inhibitors Against Clinical Isolates and Animal Study Support

VIF 抑制剂针对临床分离株的测试和动物研究支持

• 批准号: 8271416
• 负责人: BRIGITTE BEER
• 金额: $ 19.16万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-28 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8271416
• 关键词: Acute; Animals; Antiviral Agents; Applications Grants; Biological Assay; Cell fusion; Cells; Cerebrospinal Fluid; Clinical; Development; Enzymes; Fiber; Gagging; Grant; HIV; HIV Infections; HIV-1; Human; Implant; In Vitro; Infection; Lead; Lectin; Licensing; Macaca; Macaca mulatta; Massachusetts; Modeling; Monitor; Mus; New England; Peripheral Blood Mononuclear Cell; Persons; Plasma; Primates; Proteins; Public Health; RNA; Research; Research Institute; Reverse Transcriptase Polymerase Chain Reaction; SIV; Sampling; Testing; Toxic effect; Toxicity Tests; United States; Universities; Viral Load result; Viral load measurement; Virus; analog; base; efficacy testing; in vivo; inhibitor/antagonist; mouse model; neurotropic; nonhuman primate; pre-clinical; vif Gene Products

Core B will support the overall grant application by providing specific in vitro and in vivo tests of promising Vif inhibitor candidates. The first aim is to test the efficacy and toxicity of promising Vif inhibitor candidates in human PBMC-based assays using clinical HIV-1 isolates including neurotropic isolates. The second aim is to test promising Vif inhibitor candidates in the HIV-1 Hollow Fiber Mouse model. Hollow fibers filled with HIV-1 infected cells will be implanted subcutaneously and intraperitoneally into Balb/c mice. Subsequently, the mice will be treated intraperitoneally with the Vif inhibitor candidates. The mice will be monitored for acute toxicity and the fibers evaluated for efficacy against HIV-1 replication. The third aim is to perform quantitative RNA viral load determinations on rhesus macaque plasma and viral load samples in support of the SIV/primate studies at New England Primate Research Center (Project 3). For the quantitative viral load measurements, TAQMAN RT-PCR using an ABI7900 sequencer will be performed. Primers, probe, and standard are from the gag region of SIV. The relevance of this project to public health: Currently, about 1 million people are infected with HIV in the United States. In addition, 40,000 new infections are occurring each year. So far, about 500,000 persons died from complications of HIV-infection in the US. In the last 15 years, a variety of anti-HIV compounds have been developed but these compounds only control but do not cure the infection. All of the compounds currently licensed are directed against two HIV enzymes or against virus/cell fusion. Recently, the interaction of another HIV protein, Vif, with a cellular protein (APOBEC3G) was discovered. The newly identified compounds that inhibit this interaction have the potential to cure the infection with HIV-1 since Vif is required for replication in primary human cells.

核心B将通过提供有前景的VIF的特定体外和体内测试来支持整个赠款申请 抑制剂候选者。第一个目标是测试有希望的Vif抑制剂的有效性和毒性 使用临床HIV-1分离株(包括嗜神经性分离株)的基于人PBMC的检测。第二个目标是 在HIV-1中空纤维小鼠模型中测试有前景的Vif抑制剂候选药物。填充中空纤维 HIV-1感染细胞将被植入Balb/c小鼠的皮下和腹膜内。后来， 这些小鼠将接受Vif候选抑制剂的腹膜内治疗。将对这些老鼠进行监测 急性毒性和对抗HIV-1复制的有效性进行评估。第三个目标是执行 恒河猴血浆和病毒载量样本的定量RNA病毒载量测定 新英格兰灵长类研究中心的SIV/灵长类研究(项目3)。对于定量的病毒载量 测量，将使用ABI7900测序仪进行TAQMAN RT-PCR。引子、探头和 标准品来自SIV的Gag区。 该项目与公共卫生的相关性： 目前，美国约有100万人感染艾滋病毒。此外，4万个新的 感染每年都在发生。到目前为止，约有50万人死于艾滋病毒感染的并发症 在美国。在过去的15年里，已经开发出了各种抗艾滋病毒的化合物，但这些化合物 只有控制而不能治愈感染。目前获得许可的所有化合物都是针对两个 HIV酶或抗病毒/细胞融合。最近，另一种HIV蛋白Vif与细胞 蛋白质(APOBEC3G)被发现。新发现的抑制这种相互作用的化合物具有 有可能治愈HIV-1感染，因为Vif是在原代人类细胞中复制所必需的。