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

摘要-项目1 我们计划的目标是阐明HCMV调节CD34+宿主信号的分子机制 用于建立和维持病毒潜伏期和重新激活的造血祖细胞 从延迟开始。人巨细胞病毒仍然是实体器官和造血术后发病率和死亡率的重要原因 干细胞移植，尽管在诊断和治疗方面取得了进展。巨细胞病毒潜伏期很复杂，而且 建立和维持巨细胞病毒潜伏期以及病毒重新激活的信号机制 人们对此了解甚少。我们的程序已经确定了一些病毒蛋白和miRNAs来调节潜伏期 并通过靶向宿主通路重新激活。发信号通知事件的复杂性和方法 全面解决病毒潜伏期和造血问题只能通过 合作努力。利用最新的人CD34+造血干细胞体外模型和体内模型 人源化小鼠，我们的计划将解决病毒因子调节宿主的单独和联合作用 发信号。我们的项目(项目1)将解决延迟决定因素UL138在调制主机中的作用 通过其与泛素特异性蛋白酶(USP)复合体的相互作用调节的信号通路 调节STAT1和AKT信号。我们定义了UL138和WDR48以及WDR20之间的相互作用， 它们充当激活USP1、USP12和USP46的支架。我们假设UL138与宿主的相互作用 通过WDR/USP复合体协调信号来调节潜伏期和再激活之间的切换。在AIM 1，我们将确定UL138如何影响WDR48/USP复合体和功能。我们将把氨基酸映射到 与WDR48/USP复合体相互作用并产生有缺陷的重组病毒所需的UL138 对于这些互动来说。在目标2中，我们将确定UL138-WDR48/USP交互如何影响延迟和 UL138的重新激活及其在重组病毒激活STAT1和AKT中的作用及敲除 寄主因素。目标3将通过分析AKT和STAT1信号通路中的 调节STAT1和AKT信号以驱动结果的更大的调节器、蛋白质和miRNAs网络 在项目2和项目3的合作下，我们的项目将共同提供第一个 对控制宿主信号的多方面调节的全面和机械性的见解 巨细胞病毒潜伏期。我们已经确定的病毒和宿主因素的网络使我们能够定义新的宿主 以及抗病毒策略的病毒靶点，以控制HCMV潜伏期或重新激活。我们的协作方法将 提供无与伦比的洞察力，了解调节HCMV潜伏期的复杂多因素机制和 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.