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：
10216634
负责人：
JAY A NELSON
金额：
$ 26.88万
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/10216634
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项目摘要/摘要疱疹病毒miRNA靶向许多不同的细胞和病毒过程，涉及病毒免疫识别，凋亡，细胞周期调节，细胞运输以及病毒潜伏期和裂解复制。我们做了阐明HCMV miRNA的功能作用的重大进展，包括识别HCMV 调节HCMV IE1以及参与细胞周期调制的多个细胞基因的miRNA，形成病毒组装复合物和TLR2信号传导的。重要的是，我们已经观察到HCMV miRNA 协同工作以有效地下调单个基因，并且HCMV miRNA靶向多个同一细胞途径内的单个基因。最近，我们确定了几个HCMV miRNA 针对表皮生长因子受体（EGFR）信号通路的多个成员在病毒潜伏期和造血中很重要。我们已经观察到了两个的双HCMV突变靶向该途径的miRNA导致病毒重新激活增加，而另外两个病毒miRNA双重突变体在体外未能在人类祖细胞（HPC）培养系统中重新激活。最后，EGFR信号传导也证明在HPC造血中很重要。我们已经观察到HPC的单一感染双HCMV miRNA突变导致CFU-GM，CFU-GEMM和BFU-E菌落的水平降低与WT HCMV相比，形成以及骨髓抑制造血。因此，我们假设HCMV miRNA的EGFR信号调节起着关键作用在病毒潜伏期和重新激活以及造血作用中。在当前的建议中，我们将表征 EGFR HCMV miRNA靶标和EGFR miRNA靶标在病毒潜伏期和在体外CD34+ HPC中的复制。这些研究将在新发展的人胎骨肝脏中扩展和胸腺（BLT）点头scidil2RGC无效小鼠模型，能够支持潜在的HCMV感染以及从延迟（核心A）重新激活。我们将确定如何针对此途径的HCMV miRNA 项目1中病毒潜伏期的建立和维持中，具有UL133/8 EGFR调节的相间。此外，一些HCMV miRNA还靶向HCMV潜伏基因US28和 UL7。我们将研究这些miRNA在调节US28信号和US28-EGFR信号传导中扮演的角色项目3和UL7（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。