Quantitative Analysis of Cellular Signaling in Viral Latency

病毒潜伏期细胞信号传导的定量分析

• 批准号: 7421115
• 负责人: KATHRYN MILLER-JENSEN
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7421115
• 关键词: Affect; Appendix; Binding; Cells; Computational Biology; Computational Technique; Computer Simulation; Computing Methodologies; Data; Depth; Drug Design; Environment; Exhibits; Feedback; Figs - dietary; Gene Expression; Genetic Transcription; HIV; HIV-1; Heterogeneity; Individual; Infection; Jurkat Cells; Laboratories; Lead; Mediating; Modeling; Molecular; Molecular Biology Techniques; Molecular Virology; NF-kappa B; Nuclear; Patients; Pharmaceutical Preparations; Population; Public Health; Rate; Scientist; Signal Transduction; Simulate; System; T memory cell; T-Lymphocyte; Testing; Therapeutic; Toxic effect; Training; Vesnarinone; Viral; Virus; Virus Latency; base; design; experience; latent infection; mathematical model; novel; promoter; purge; research study; therapy design

DESCRIPTION (provided by applicant): Latent HIV is the most significant barrier to complete eradication of HIV from a patient. However, the molecular basis of latency remains unknown. Using a retroviral model of the HIV-1 Tat-mediated positive feedback loop, the laboratories of Profs. Schaffer (sponsor) and Arkin (co-sponsor) showed that clonal populations of infected Jurkat cells with a single viral integration can either rapidly initiate viral gene expression or exhibit long periods of low gene expression analogous to a latent infection. We hypothesize that these long stochastic transcriptional and translational delays could maintain the virus in an inactive state long enough for the host T cell to convert to a memory T cell and thereby solidify the virus into a latent state, a fundamentally new hypothesis for how HIV latency is established. I am using experimental and computational techniques to probe how NF-kB, a key transcriptional regulator of the HIV LTR promoter, influences stochastic gene expression and latency. My central hypotheses are 1) that fluctuations in the interaction of NF-kB at the HIV LTR in single T cells alters the basal transcription rate and leads to the activation or latency decision; and 2) that with an appropriate strength and duration of NF-kB activation, it is possible to purge the latent pool without toxicity. These hypotheses will be tested through a set of experiments organized into two specific aims. In Specific Aim I, I will determine if cell-to-cell heterogeneity in NF-kB interactions at the HIV LTR promoter contributes to stochasticity in HIV gene expression. To do this, I will collect quantitative data using molecular biology techniques to perturb NF-kB LTR binding and nuclear concentrations in infected Jurkat cells, and then use these data to computationally simulate the HIV latency decision. In Specific Aim II, I will use this computational model of NF-kB-dependent HIV gene expression to design therapeutic anti-latency strategies, and then experimentally test efficacy. This blend of molecular virology and computational biology promises to make progress in designing therapies for a major biomedical problem. PUBLIC HEALTH RELEVANCE: Latent HIV, a pool of replication-competent virus that "hides" in host cells, is the significant barrier curing HIV-infected patients, however, scientists still do not know how latency is established. I am building a mathematical model to investigate how HIV can lead to active infection in some cells and latent infection in other cells. I will then use this model to help design anti-latency drug strategies to purge latent infections from a cell population.

描述（由申请人提供）：潜伏的HIV是彻底根除患者体内HIV的最大障碍。然而，潜伏的分子基础尚不清楚。利用HIV-1 tat介导的正反馈回路的逆转录病毒模型，Schaffer（赞助者）和Arkin（共同赞助者）表明，具有单一病毒整合的受感染Jurkat细胞克隆群体可以迅速启动病毒基因表达或表现出类似于潜伏感染的长时间低基因表达。我们假设，这些长时间的随机转录和翻译延迟可以使病毒保持在非活性状态，使宿主T细胞转化为记忆T细胞，从而使病毒固化为潜伏状态，这是一个关于HIV潜伏期如何建立的全新假设。我正在使用实验和计算技术来探索NF-kB （HIV LTR启动子的关键转录调节因子）如何影响随机基因表达和潜伏期。我的主要假设是1)单个T细胞中NF-kB在HIV LTR处相互作用的波动改变了基础转录率并导致激活或潜伏期的决定；2)通过适当的NF-kB激活强度和持续时间，可以清除潜伏池而不产生毒性。这些假设将通过分为两个具体目标的一系列实验来检验。在Specific Aim I中，我将确定HIV LTR启动子上NF-kB相互作用的细胞间异质性是否有助于HIV基因表达的随机性。为此，我将使用分子生物学技术收集定量数据来干扰NF-kB LTR结合和受感染Jurkat细胞中的核浓度，然后使用这些数据来计算模拟HIV潜伏期决策。在Specific Aim II中，我将使用这种nf - kb依赖性HIV基因表达的计算模型来设计治疗性抗潜伏期策略，然后通过实验测试效果。这种分子病毒学和计算生物学的结合有望在设计治疗重大生物医学问题的方法方面取得进展。