权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

HCMV miRNA regulation of host cell signaling in viral latency and hematopoiesis

HCMV miRNA 对病毒潜伏期和造血过程中宿主细胞信号传导的调节

基本信息

批准号：
9980281
负责人：
JAY A NELSON
金额：
$ 27.08万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-15 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9980281
关键词：
Apoptosis Applications Grants BFU-E BLT mice CD34 gene Cell Culture System Cell Cycle Cell Cycle Regulation Cells Clinical Collaborations Complex Cytomegalovirus Data EGF gene Epidermal Growth Factor Receptor Gene Expression Generations Genes Genetic Transcription Goals Hematopoiesis Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Herpesviridae Human Immune In Vitro Individual Infection Interphase Knock-out Knockout Mice Liver Maintenance Mediating MicroRNAs Molecular Morbidity - disease rate Mutation Myelosuppression Organ Transplantation Pathway interactions Patients Play Process Program Research Project Grants Receptor Signaling Recombinants Regulation Reporter Role Signal Pathway Signal Transduction Small RNA Solid TLR2 gene Thymus Gland Transplantation Viral Viral Physiology Virus Virus Assembly Virus Latency Virus Replication Work bone cellular targeting cytomegalovirus receptor fetal human fetal bone marrow in vivo lytic replication member mortality mouse model mutant reactivation from latency small hairpin RNA stem cells trafficking

项目摘要

PROJECT 2 PROJECT SUMMARY/ABSTRACT Herpesvirus miRNAs target many different cellular and viral processes involved in viral immune recognition, apoptosis, cell cycle regulation, cellular trafficking as well as viral latency and lytic replication. We have made significant advances elucidating functional roles for HCMV miRNAs including the identification of HCMV miRNAs that regulate HCMV IE1 as well as multiple cellular genes involved in cell cycle modulation, formation of the viral assembly complex, and TLR2 signaling. Importantly, we have observed that the HCMV miRNAs work coordinately to efficiently down-regulate individual genes and that the HCMV miRNAs target multiple individual genes within the same cellular pathways. Recently, we have determined that several HCMV miRNAs target multiple members of the Epidermal Growth Factor Receptor (EGFR) signaling pathways that are important in viral latency and hematopoiesis. We have observed that a double HCMV mutation of two of the miRNAs targeting this pathway results in an increase in viral reactivation while two other viral miRNA double mutants fail to reactivate in a human progenitor cell (HPC) culture system in vitro. Lastly, EGFR signaling has also been shown to be important in HPC hematopoiesis. We have observed that infection of HPCs with single and double HCMV miRNA mutations results in reduced levels of CFU-GM, CFU-GEMM and BFU-E colony formation as well as myelosuppression in comparison to WT HCMV, suggesting a role for the miRNAs in hematopoiesis. Therefore, we hypothesize that HCMV miRNA regulation of EGFR signaling plays a critical role in viral latency and reactivation as well as hematopoiesis. In the current proposal we will characterize the EGFR HCMV miRNA targetome and the functional relevance of the EGFR miRNA targets for viral latency and replication in CD34+ HPC in vitro. These studies will be extended in a newly developed human fetal bone, liver and thymus (BLT) NOD-scidIL2Rgc null mouse model that is able to support latent HCMV infection as well as reactivation from latency (Core A). We will determine how the HCMV miRNAs that target this pathway interphase with UL133/8 EGFR regulation in the establishment and maintenance of viral latency in Project 1. Additionally, several of the HCMV miRNAs also target genes activated by the HCMV latency gene US28 and UL7. We will examine the role that these miRNAs play in regulating US28 signaling and US28-EGFR signaling cross-talk in Project 3 and UL7 (Project 4). Lastly, we will determine the contribution of the HCMV EGFR miRNAs in hematopoiesis in comparison to results of WT HCMV in Project 5.

项目2项目概要/摘要疱疹病毒miRNAs靶向参与病毒免疫识别的许多不同的细胞和病毒过程，细胞凋亡、细胞周期调节、细胞运输以及病毒潜伏和裂解复制。我们取得了阐明HCMV miRNA功能作用的重要进展，包括HCMV的鉴定调节HCMV IE 1以及参与细胞周期调节、形成和/或凋亡的多个细胞基因的miRNA 和TLR 2信号传导。重要的是，我们已经观察到HCMV miRNAs 协同工作，有效地下调个别基因，HCMV miRNAs靶向多个同一细胞通路中的单个基因。最近，我们已经确定了几个HCMV miRNAs，靶向表皮生长因子受体（EGFR）信号通路的多个成员，在病毒潜伏期和造血中起重要作用。我们已经观察到，两个HCMV基因的双重突变，靶向该途径的miRNA导致病毒再活化增加，而另外两种病毒miRNA则加倍。突变体在体外人祖细胞（HPC）培养系统中不能再活化。最后，EGFR信号传导具有也被证明在HPC造血中是重要的。我们已经观察到， HCMV miRNA双突变导致CFU-GM、CFU-GEMM和BFU-E集落水平降低与野生型HCMV相比，miRNAs在HCMV的形成以及骨髓抑制中的作用，表明miRNAs在HCMV的形成中的作用。造血因此，我们假设HCMV miRNA对EGFR信号的调节在EGFR的表达中起着关键作用。在病毒潜伏期和再活化以及造血方面。在目前的提案中，我们将描述 EGFR HCMV miRNA靶向组和EGFR miRNA靶点对病毒潜伏期的功能相关性，在体外CD 34 + HPC中复制。这些研究将在一个新开发的人类胎儿骨，肝脏和胸腺（BLT）NOD-scidIL 2 Rgc无效小鼠模型，其能够支持潜伏性HCMV感染以及从延迟重新激活（核心A）。我们将确定靶向这一通路的HCMV miRNAs 在项目1中，在病毒潜伏期的建立和维持中与UL 133/8 EGFR调节的间期。此外，几种HCMV miRNA还靶向由HCMV潜伏基因US 28激活的基因， UL7我们将研究这些miRNAs在调节US 28信号和US 28-EGFR信号中的作用。项目3和UL 7（项目4）中的串扰。最后，我们将确定HCMV EGFR的贡献，与项目5中WT HCMV的结果相比，造血中的miRNA。