Manipulation of host factors that promote HCMV latency

项目摘要/摘要--O‘Connor，Christine M. 人巨细胞病毒(HCMV)是一种广泛传播的病原体，感染大多数人 美国的人口。这种病毒对发展中国家的发展构成了重大威胁 胎儿以及缺乏有效免疫系统的儿童和成人，通常 导致严重的疾病和死亡。一旦个人感染了巨细胞病毒， 病毒终生与宿主在一起，在造血系统中处于潜伏或静止状态 车厢。在严重的免疫应激时期，病毒会重新激活到它的 活跃状态，允许传播和随后的疾病。除了 免疫幼稚和血清阴性的器官移植受者，初次感染 人巨细胞病毒很少导致疾病，但正是重新激活导致了显著的 并发症。因此，为了预防巨细胞病毒相关疾病，我们必须获得一个完整的 了解病毒潜伏期和重新激活。少数几个编码的基因之一 潜伏期是四种病毒G蛋白偶联受体(GPCRs)之一，US28。我们 以前已经表明，建立和维护需要US28 潜伏期和US28介导的信号转导促成了这些效应。我们还有 发现US28调节特定细胞靶标的表达，这些靶标调节 主要的即刻早期促进剂(MIEP)，潜伏期到溶血期开关的主要调节者。 因此，我们假设US28调节特定的宿主信号通路以 调节MIEP的转录沉默以促进HCMV潜伏期。要探索这一点 假设，我们将利用新的方法和我们的28美元军火库- 特定重组体，以及体外和体外潜伏期模型系统。在AIM 1，我们将定义US28介导的AP-1衰减的潜在机制 通过检测上游转录因子在潜伏期与MIEP结合 调节AP-1的信号通路。在目标2中，我们将确定招聘的因素 YY1在潜伏期通过评估US28调节的信号通路和 促进这种抑制转录因子结合的因子。总而言之， 本文提出的实验将使我们更好地理解S的生物学 在潜伏期内发挥作用，并将为未来的研究开发新的 专门针对人巨细胞病毒感染潜伏宿主的治疗。

PROJECT SUMMARY/ABSTRACT – O’CONNOR, CHRISTINE M. Human cytomegalovirus (HCMV) is a wide-spread pathogen, infecting the majority of the population in the United States. This virus poses a significant threat to developing fetuses as well as to children and adults who lack a competent immune system, often causing severe disease and mortality. Once individuals acquire an HCMV infection, the virus remains with the host for life, in a latent or quiescent state in the hematopoietic compartment. During times of severe immunological stress, the virus reactivates to its active state, allowing for dissemination and subsequent disease. With the exception of the immuno-naïve and sero-negative organ transplant recipients, primary infection with HCMV rarely causes disease, but rather it is reactivation that leads to significant complications. Thus, to prevent HCMV-associated disease, we must gain a complete understanding of viral latency and reactivation. One of a handful of genes encoded during latency is one of the four viral G protein-coupled receptors (GPCRs), US28. We have shown previously that US28 is required for the establishment and maintenance latency and that US28-mediated signaling contributes to these effects. We have also found that US28 modulates the expression of specific cellular targets that regulate the Major Immediate Early Promoter (MIEP), a master regulator in the latent-to-lytic switch. Therefore, we hypothesize that US28 modulates specific host signaling pathways to regulate transcriptional silencing of the MIEP to facilitate HCMV latency. To explore this hypothesis, we will take advantage novel approaches coupled with our arsenal of US28- specific recombinants, as well as both in vitro and ex vivo latency model systems. In Aim 1, we will define mechanisms underlying US28-mediated attenuation of AP-1 transcription factor binding to the MIEP during latency by examining the upstream signaling pathways that regulate AP-1. In Aim 2, we will determine the factors that recruit YY1 to the MIEP during latency by assessing US28-regulated signaling pathways and factors that promote the binding of this repressive transcription factor. In sum, the experiments proposed herein will lead to a greater understanding of US28’s biological functions during latency and will lay the foundation for future studies to develop novel therapeutics specifically targeting the latent reservoir of HCMV infection.