Virus-host interactions regulating innate signaling for human cytomegalovirus latency

病毒-宿主相互作用调节人类巨细胞病毒潜伏期的先天信号

基本信息

批准号：
10565926
负责人：
Felicia D Goodrum
金额：
$ 37.68万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-07 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10565926
关键词：
Address Amino Acids Biology CD34 gene Cells Cercopithecine Herpesvirus 1 Complex Complication Cytomegalovirus Cytomegalovirus Infections DNA Damage DNA biosynthesis Data Defective Viruses Dependence Diagnostic Disease Endothelial Cells Epidermal Growth Factor Receptor Genes Genome Goals Growth Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human Immune Immunoprecipitation In Vitro Individual Infection Integration Host Factors Interferons Life MEKs Maintenance Maps Mass Spectrum Analysis Mediating Molecular Morbidity - disease rate Organ PI3K/AKT Pathway interactions Positioning Attribute Proteins Proto-Oncogene Proteins c-akt Regulation Reporting Research Role STAT1 gene Signal Transduction Solid Stem cell transplant Stimulus Switching Complex Therapeutic Transcript Undifferentiated Viral Viral Genes Virus Virus Latency Virus Replication WD Repeat Work attenuation cell type cofactor collaborative approach humanized mouse in vitro Model in vivo in vivo Model insight knock-down latent infection mortality mouse model mutant novel novel virus organ transplant recipient prevent programs protein transport reactivation from latency recombinant virus response scaffold stem cells trafficking transcription factor ubiquitin-specific protease viral DNA virus host interaction

项目摘要

PROJECT SUMMARY The goal of our research 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 regulating the establishment and maintenance of HCMV latency, as well as for reactivation of HCMV, are poorly understood. We have identified a locus of viral genes in the ULb’ region of the genome that coordinates the expression of four genes, UL133, UL135, UL136, and UL138, from polycistronic transcripts. Using state-of-the art in vitro models in human CD34+ HPCs and in vivo models in humanized mice, we will define virus-host interactions modulating host signaling for the establishment of latency. Our preliminary data show that the latency determinant, UL138, interacts with host WD Repeat containing protein 48 (WDR48), which serves as a scaffold to activate ubiquitin specific protease, USP1, USP12, and USP46. WDR48-USP1 complexes regulate STAT1 and AKT signaling and we demonstrate that UL138 directs the activity of WDR48-USP1 to induce the activation of STAT1. We further show that USP1 activity it important for the establishment of a latent infection, such that when USP1 is inhibited, the virus replicates in the absence of a replication stimulus. We hypothesize that UL138 interaction directs WDR48-USP complexes to regulate innate signaling to suppress virus replication to prevent 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. We will distinguish roles of the UL138-WDR48/USP1 interactions from that of UL138-EGFR interactions. Further, we have shown that UL138 sustains AKT signaling, at least in part through an interaction with EGFR, but interaction with USP12 and USP46 may provide additional avenues to the regulation of AKT. Aim 2 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. This project will provide the 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.

项目概要我们研究项目的目标是阐明 HCMV 调节宿主的分子机制 CD34+ 造血祖细胞 (HPC) 中的信号传导用于病毒的建立和维持 HCMV 的潜伏期和潜伏期的重新激活仍然是固体治疗后发病和死亡的重要原因。尽管 HCMV 的诊断和治疗取得了进展，但器官和造血干细胞移植仍然存在。潜伏期复杂，调节HCMV建立和维持的信号机制我们对 HCMV 的潜伏期以及重新激活知之甚少。基因组的 ULb' 区域协调四个基因的表达：UL133、UL135、UL136 和 UL138，来自多顺反子转录本，在人类 CD34+ HPC 和体内使用最先进的体外模型。在人源化小鼠模型中，我们将定义病毒与宿主的相互作用，调节宿主信号传导我们的初步数据表明，延迟决定因素 UL138 与主机相互作用。 WD Repeat 含有蛋白 48 (WDR48)，可作为激活泛素特异性蛋白酶的支架， USP1、USP12 和 WDR48-USP1 复合物调节 STAT1 和 AKT 信号传导，我们证明了这一点。 UL138 指导 WDR48-USP1 的活性诱导 STAT1 的激活。它的活性对于潜伏感染的建立很重要，因此当 USP1 被抑制时，病毒我们勇敢地说，UL138 相互作用指导 WDR48-USP。在目标 1 中，我们将通过调节先天信号传导来抑制病毒复制以防止病毒重新激活。确定 UL138 如何影响 WDR48/USP 复合物和功能我们将绘制 UL138 中的氨基酸图谱。与 WDR48/USP 复合物相互作用所需的，并产生有缺陷的重组病毒我们将区分 UL138-WDR48/USP1 相互作用与 UL138-EGFR 的作用。此外，我们还表明 UL138 至少部分通过相互作用维持 AKT 信号传导。与 EGFR 的相互作用，但与 USP12 和 USP46 的相互作用可能为 AKT 的调节提供额外的途径。目标 2 将确定 UL138-WDR48/USP 相互作用如何影响延迟和重新激活以及 UL138 使用重组病毒和宿主因子敲低激活 STAT1 和 AKT。将为宿主信号传导的多方面调控提供全面和机械的见解我们已经确定的病毒和宿主因素网络使我们能够控制 HCMV 潜伏期。定义新的宿主和病毒靶标，用于控制 HCMV 潜伏期或再激活的抗病毒策略。