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.

项目摘要我们的研究计划的目的是阐明HCMV调节宿主的分子机制 CD34+造血祖细胞（HPC）中的信号传导，以建立和维持病毒潜伏期和延迟重新激活。 HCMV仍然是固体后发病率和死亡率的重要原因器官和造血干细胞移植目的地诊断和治疗的进步。 HCMV 潜伏期很复杂，并且信号机制可以预期建立和维护HCMV 延迟以及HCMV的重新激活，知之甚少。我们已经确定了病毒基因的基因座基因组的ULB区域，该区域协调了四个基因的表达，UL133，UL135，UL136和 UL138，来自多元转录本。在人CD34+ HPC和体内使用最新的体外模型人性化小鼠中的模型，我们将定义病毒宿主相互作用调节宿主信号的信号建立潜伏期。我们的初步数据表明，延迟确定器UL138与主机交互 WD重复含有蛋白48（WDR48），作为激活泛素特异性蛋白的支架， USP1，USP12和USP46。 WDR48-USP1复合物调节STAT1和AKT信号，我们证明了 UL138指示WDR48-USP1的活性诱导STAT1的激活。我们进一步表明USP1 活动对于建立潜在感染很重要，因此当抑制USP1时，病毒在没有复制刺激的情况下复制。我们假设UL138相互作用指导WDR48-USP 复合物调节先天信号以抑制病毒复制以防止重新激活。在AIM 1中，我们将确定UL138如何影响WDR48/USP复合物和功能。我们将在UL138中绘制氨基酸与WDR48/USP复合物相互作用并生成重组病毒缺陷所必需的这些相互作用。我们将区分UL138-WDR48/USP1相互作用的角色与UL138-EGFR 互动。此外，我们已经证明UL138自杀Akt信号至少部分通过相互作用使用EGFR，但是与USP12和USP46的相互作用可能会为AKT的调节提供其他途径。 AIM 2将确定UL138-WDR48/USP相互作用如何影响潜伏期和重新激活以及 UL138使用重组病毒激活STAT1和AKT并敲低宿主因子。这个项目将为主机信号的多方面调节提供全面和机械的见解 HCMV延迟的控制。我们已经确定的病毒和宿主因素网络将我们定位为定义新的宿主和病毒靶标，以控制HCMV潜伏期或重新激活的抗病毒策略。