HCMV UL133/8 regulation of host cell signaling in viral latency and reactivation

摘要-项目1 本项目的目标是阐明HCMV调节宿主CD 34+细胞信号转导的分子机制。 造血祖细胞（HPC）用于建立和维持病毒潜伏和再活化 从latency。HCMV仍然是实体器官和造血干细胞移植后发病和死亡的重要原因。 干细胞移植尽管在诊断和治疗方面取得了进展。HCMV潜伏期是复杂的， HCMV潜伏期的建立和维持以及病毒再活化的信号传导机制 我们对此知之甚少。我们的项目已经确定了许多调节潜伏期的病毒蛋白和miRNAs 并通过靶向宿主途径重新激活。信号事件和方法的复杂性， 全面解决病毒潜伏期和造血问题只能通过 合作的努力。在人CD 34 + HPC中使用最先进的体外模型，在人CD 34 + HPC中使用最先进的体内模型， 人源化小鼠，我们的计划将解决病毒因子调节宿主的个体和组合作用， 发信号。我们的项目（项目1）将探讨潜伏期决定因子UL 138在调节宿主细胞中的作用。 通过其与泛素特异性蛋白酶（USP）复合物的相互作用调节信号通路， 调节STAT 1和AKT信号传导。我们已经定义了UL 138和WDR 48和WDR 20之间的相互作用， 其充当支架以激活USP 1、USP 12和USP 46。我们假设UL 138与宿主的相互作用 WDR/USP复合物协调信号传导以调节潜伏期和再激活之间的转换。在Aim中 1，我们将确定UL 138如何影响WDR 48/USP复合物和功能。我们将绘制出 与WDR 48/USP复合物相互作用所需的UL 138，并产生缺陷型重组病毒 对于这些互动。在目标2中，我们将确定UL 138-WDR 48/USP相互作用如何影响延迟， 使用重组病毒和基因敲低， 宿主因素目的3将通过分析一种新的信号转导途径，扩展我们对STAT 1和AKT信号转导调控的理解。 更大的调节因子、蛋白质和miRNA网络，调节STAT 1和AKT信号传导以驱动结果 与项目2和项目3合作。总的来说，我们的项目将提供第一个 全面和机械的见解，多方面的调节宿主信号的控制， HCMV潜伏期。我们已经确定的病毒和宿主因子网络使我们能够定义新的宿主 以及用于控制HCMV潜伏期或再活化的抗病毒策略的病毒靶点。我们的合作方式将 为调节HCMV潜伏期的复杂多因素机制提供了无与伦比的见解， 重新激活

SUMMARY – PROJECT 1 The goal of our Program is to elucidate molecular mechanisms by which HCMV regulates host signaling in CD34+ hematopoietic progenitor cells (HPCs) for the establishment and maintenance of viral latency and reactivation from latency. HCMV remains a significant cause of morbidity and mortality after solid organ and hematopoietic stem cell transplantation despite advances in diagnostics and therapeutics. HCMV latency is complex and the signaling mechanisms for establishment and maintenance of HCMV latency, as well as for reactivation of virus are poorly understood. Our program has identified a number of viral proteins and miRNAs that regulate latency and reactivation by targeting host pathways. The complexity of signaling events and approaches to comprehensively address questions on viral latency and hematopoiesis can only be achieved through a collaborative effort. Using state-of-the art in vitro models in human CD34+ HPCs and in vivo models in humanized mice, our Program will address the individual and combined roles of viral factors modulating host signaling. Our Project (Project 1) will address the roles of the latency determinant UL138 in modulating host signaling pathways regulated through its interactions with ubiquitin specific protease (USP) complexes that regulate STAT1 and AKT signaling. We have defined interactions between UL138 and WDR48 and WDR20, which serve as a scaffold to activate USP1, USP12, and USP46. We hypothesize that UL138-host interactions with WDR/USP complexes coordinate signaling to regulate the switch between latency and reactivation. In Aim 1, we will determine how UL138 impacts WDR48/USP complexes and function. We will map the amino acids in UL138 that are required for interaction with WDR48/USP complexes and generate recombinant viruses defective for these interactions. In Aim 2, we will determine how UL138-WDR48/USP interactions impact latency and reactivation and the role of UL138 in activating STAT1 and AKT using recombinant viruses and knockdown of host factors. Aim 3 will expand our understanding of the regulation of STAT1 and AKT signaling by analyzing a greater network of regulators, proteins and miRNAs, that regulate STAT1 and AKT signaling to drive outcomes of infection in collaboration with Project 2 and Project 3. Collectively, our projects will provide the first comprehensive and mechanistic insights into the multi-faceted regulation of host signaling for the control of HCMV latency. The network of viral and host factors we have identified uniquely position us to define novel host and viral targets for antiviral strategies to control HCMV latency or reactivation. Our collaborative approach will provide unparalleled insights into the complex multi-factorial mechanisms regulating HCMV latency and reactivation.