Control of Human Cytomegalovirus

描述(由申请人提供)： 尽管抗病毒治疗取得了进展，但人类巨细胞病毒(HCMV)持续感染着一半以上的退伍军人，并在这一人群中的相当一部分人中导致疾病和死亡。需要采取新的战略来减轻由人巨细胞病毒感染造成的疾病负担。HCMV mie增强子/启动子是病毒的裂解开关。它被打开以启动病毒复制周期，并在病毒潜伏期间关闭。有关控制HCMV裂解开关的方法的知识有可能为预防病毒复制的新疗法的开发提供信息。我们的实验室处于确定控制机制的调查的前沿。这些早期的结果使我们假设，在急性产生性感染和再激活感染中，控制HCMV裂解开关激活的转录调控等级和顺式调控码在不同的细胞类型中是不同的。在人类多能性N-Tera2细胞(NT2)中的初步数据表明，通过不同的信号和多种调控途径的组合来实现HCMV mie的重新激活是最好的。既需要进一步阐明所涉及的线索和调控途径，也需要在树突状单核细胞(D/M)前体的HCMV潜伏期模型中评估这些发现。这些结果将首次阐明控制HCMV裂解开关重新激活的分子机制。在急性感染的许可成纤维细胞中的初步数据揭示了一种新的调控机制，它在感染的后即刻-早期阶段发挥非典型的作用，并与另一种尚未确定的调控机制一起作用。填补关于这一调控机制的作用、机制和目的的知识空白有望改变解释MIE增强子如何管理HCMV复制的范式。这个项目的具体目的是描述特定的转录因子途径和顺式调控码在以下几个方面的不同作用：1)急性感染的许可成纤维细胞、内皮细胞、上皮细胞和髓系细胞；2)静止感染的NT2和D/M前体细胞。该研究计划将利用创新的方法和工具。先进技术和最先进方法的使用使得在生物进口的不同细胞环境中以无与伦比的深度详细描述这些途径是可行的。卓越的研究环境和成熟的研究团队与爱荷华大学的中央显微镜研究、蛋白质组学和流式细胞术设施很好地结合在一起，是实施这一计划的理想选择。病毒调控机制的发现可以在病毒开始之前阻止HCMV基因的表达和复制，这可能为抗病毒药物的开发提供了新的方向，目标是改善退伍军人的健康。

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) persistently infects over half of the Veteran population and causes disease and death in a sizeable segment of this population despite advances in antiviral therapy. Novel strategies to mitigate the disease burden resulting from HCMV infection are needed. The HCMV MIE enhancer/promoter serves as the lytic switch for the virus. It is turned on to initiate the viral replicative cycle and turned off during viral latency. Knowledge about the ways in which the HCMV MIE lytic switch is controlled has the potential to inform the development of new therapies to pre-empt viral replication. Our laboratory is at the forefront of investigations into defining the control mechanisms. These early results have led us to hypothesize that the transcriptional regulatory hierarchies and cis-regulatory codes in control of HCMV MIE lytic switch activation differ between cell types in acutely productive and reactivation infections. Preliminary data in human pluripotent N-Tera2 cells (NT2) suggest that HCMV MIE reactivation is best achieved by a combination of different cues and multiple regulatory pathways. There is both a need to further elucidate the cues and regulatory pathways involved and a need to evaluate these findings in a HCMV latency model in dendritic- monocytic cell (D/M) precursors. These results will be the first to elucidate the molecular mechanisms controlling HCMV MIE lytic switch reactivation. Preliminary data in acutely infected permissive fibroblasts reveal a novel regulatory mechanism which functions atypically in the post-immediate-early phase of infection and acts together with another yet unidentified regulatory mechanism. Filling in the gaps in knowledge about action, mechanism, and purpose of this regulatory mechanism is anticipated to shift paradigm in explaining how the MIE enhancer governs HCMV replication. The specific aims of this project are to delineate the differential roles of specific transcription factor pathways and cis-regulatory codes in HCMV MIE lytic switch activation in: 1) acutely infected permissive fibroblast, endothelial, epithelial, and myeloid cells; and 2) quiescently infected NT2 and D/M precursors. The research plan will draw on both innovative approaches and tools. The use of advanced technology and state-of-the-art methods makes feasible the detailing of these pathways at unparalleled depth in diverse cellular settings of biological import. An outstanding research environment and an accomplished research team that interfaces well with the Central Microscopy Research, Proteomics, and Flow Cytometry facilities at the University of Iowa are ideally suited for carrying out this plan. The discovery of viral regulatory mechanisms that can be disrupted to stop HCMV gene expression and replication before it begins potentially provides a new direction in antiviral drug development, with the goal of improving Veterans' health.