Activity of novel drug candidates in primary cell models of HIV latency

HIV潜伏原代细胞模型中新候选药物的活性

• 批准号: 8377507
• 负责人: VICENTE PLANELLES
• 金额: $ 40.97万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8377507
• 关键词: Animals; Biology; CD4 Positive T Lymphocytes; Cell Culture System; Cell Line; Cell model; Cells; Characteristics; Development; Drug Combinations; Drug Compounding; Frequencies; Future; Genetic Transcription; Goals; HIV; HIV-1; Human; In Vitro; Infection; Instruction; Knowledge; Latent Virus; Modeling; Mus; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Primary Cell Cultures; Probability; Screening procedure; Site; Stimulus; System; T memory cell; Testing; Therapeutic; Toxic effect; Viral; base; biological systems; cell type; cellular targeting; collaboratory; cytokine; drug candidate; in vivo; macrophage; nonhuman primate; novel; peripheral blood; programs; small molecule; success

Latently infected, quiescent CD4 T cells represent a central obstacle to eradication of HIV-1. Major hurdles are the low frequencies of latently infected cells in peripheral blood and the lack of known phenotypic markers that can distinguish them from uninfected ones. These impediments have prompted the development of in vitro cell models of latency, which. Such models allow manipulation of cellular and viral characteristics to gain a mechanistic understanding of how latency is established and regulated. However, no single experimental system of HIV latency is perceived to completely recapitulate the biology ofthe latent viral reservoir in vivo. This is mainly because (a) the mechanisms for establishment of latency by HIV are multiple; and (b) the types of cells harboring latent reservoirs are also multiple. A wealth of knowledge has been gained regarding how specific stimuli (e.g. activation/differentiation, homeostatic proliferation, cytokines) and viral factors (e.g. integration site, Tat-driven expression) influence the dynamics of latent reservoirs in experimental systems. Project 2.2 will perform "secondary level" characterization of novel small molecule compounds and combinations that have been identified to induce HIV transcription through primary screening by Dr. Hazuda's group in Project 2.1. In Aims 1 and 2, we propose to use a panel of wellcharacterized primary cell systems and one Jurkat-based cell line to validate candidate drug compounds. A third Aim will be the development of additional primary cell models to enable the study of HIV-1 latency in other relevant cell types not previously explored ex vivo (e.g. transitional memory T cells and macrophages). Drugs and combinations that are active in at least one ofthe primary culture systems, without showing overt toxicity or induction of cellular activation/proliferation, will be moved to subsequent phases of characterization and development within this Collaborative program. This will include detailed mechanistic studies (Objective 1), testing for activity in humanized mice and non-human primates (Objective 3) and in patient cells ex vivo (Objective 4). Our ultimate goal is to identify novel drugs and combinations that will have a high probability for success in activating HIV latent viruses when applied in patients.

静态感染的静态CD4 T细胞代表了根除HIV-1的核心障碍。主要的障碍是外周血液中受感染细胞的低频，缺乏可将其与未感染的表型标记区分开的已知表型标记。这些障碍促使了潜伏期的体外细胞模型的发展。这样的模型允许操纵细胞和病毒特征，从而对如何建立和调节潜伏期有一种机械理解。然而，没有发现单一的艾滋病毒潜伏期实验系统可以完全概括体内潜在病毒储存的生物学。这主要是因为（a）通过艾滋病毒建立潜伏期的机制是多重的； （b）带有潜在储层的细胞的类型也是多种的。关于特定刺激（例如激活/分化，稳态增殖，细胞因子）和病毒因子（例如，整合位点，TAT驱动的表达）如何影响实验系统中潜在储层的动力学。项目2.2将对新型的小分子化合物和组合进行“次要水平”表征，这些化合物和组合已通过Hazuda博士组在项目2.1的主要筛查中诱导HIV转录。在AIMS 1和2中，我们建议使用一组良好的原代细胞系统和一个基于Jurkat的细胞系来验证候选药物化合物。第三个目的是开发其他主要细胞模型，以使其他相关细胞类型中的HIV-1潜伏期可以研究（例如过渡记忆T细胞和巨噬细胞）。至少在主要培养系统中活跃的药物和组合，而没有显示出明显的毒性或细胞激活/增殖的诱导，将移至此协作计划中的表征和发展的后续阶段。这将包括详细的机械研究（目标1），对人道化小鼠和非人类灵长类动物的活性测试（目标3）以及在体内的患者细胞（目标4）。我们的最终目标是鉴定新的药物和组合，这些药物和组合将在激活患者时成功激活艾滋病毒潜伏病毒的可能性很大。