Molecular Aspects of Cytomegalovirus Latency

巨细胞病毒潜伏期的分子方面

• 批准号: 8606384
• 负责人: JAY A NELSON
• 金额: $ 49.96万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606384
• 关键词: Antiviral Agents; Apoptosis; Base Pairing; CD34 gene; Cell Culture System; Cell Cycle Regulation; Cells; Complement Factor B; Congenital Abnormality; Cytomegalovirus; Cytomegalovirus Infections; Funding; Gene Expression; Genes; Goals; Herpesviridae; Human; Immune; In Vitro; Individual; Infection; Interleukin-1; Knock-out; Knockout Mice; Latent Virus; Maintenance; MicroRNAs; Molecular; Mus; Mutation; Nuclear; Pathway interactions; Patients; Play; Process; Proteins; RNA; Regulation; Repression; Role; Signal Pathway; Small Interfering RNA; Stem cells; System; TNF gene; Time; Tumor Necrosis Factor-alpha; Viral; Viral Genome; Virus; Virus Latency; Work; cell growth regulation; cytokine; human TNF protein; inhibitor/antagonist; latent persistent infection; lytic replication; mouse model; mutant; pathogen; public health relevance; reactivation from latency; response; small hairpin RNA

DESCRIPTION (provided by applicant): The long-term goal of this project is to develop an understanding of the cellular and molecular mechanisms of human cytomegalovirus (HCMV) persistence in the host. HCMV is a significant pathogen in immune compromised patients and the leading viral cause of congenital defects. Our group and others have shown that HCMV and other herpesviruses encode multiple microRNAs (miRNAs) that are small 21-24 base pair (bp) single-stranded RNA species that regulate gene expression through post-transcriptional mechanisms. Herpesvirus miRNAs have been shown to target many different cellular and viral processes involved in viral immune recognition, apoptosis, cell cycle regulation, as well as viral latency and lytic replication. During the previous funding period our group has made significant advances elucidating functional roles for HCMV miRNAs. We have demonstrated that one of the HCMV miRNAs efficiently targets and reduces expression the IE72 protein that significantly reduces viral replication suggesting a role for maintenance of latent virus. Secondly, we have also found that the HCMV miRNAs coordinately work together to efficiently down-regulate individual genes that may explain why individual miRNA knockouts in the viral genome do not show phenotypic effects. Additionally, the HCMV miRNAs appear to also target multiple individual cellular genes in the same cellular pathways including two potential antiviral cytokines IL-1 and TNF-¿ as well as the NF?B inhibitor IkBa . Lastly, we have observed that a double HCMV miRNA mutation 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. Interestingly, one of the HCMV miRNAs in the double-mutants that fail to reactivate targets two NF?B activators suggesting that NFkB activation is deleterious for reactivation of virus. Therefore, in the current proposal we will fully characterize the IL-1 and TNF-¿ signaling pathways targeted by the HCMV miRNAs and their functional relevance for viral latency and replication in a CD34+ HPC. We will also examine the function of HCMV miRNA targeting of the IL-1 and TNF-¿ signaling pathways during latency and reactivation in a newly developed human CD34+-engrafted NOD-scidIL2Rgc null mouse model that is able to support latent HCMV infection as well as reactivation from latency. We hypothesize that HCMV miRNA repression of viral replication is related to NFkB activation through the IL-1 and TNF-¿ pathways.

描述（由适用提供）：该项目的长期目标是对宿主中人类巨细胞病毒（HCMV）持久性的细胞和分子机制有一种理解。 HCMV是免疫组合患者的重要病原体，也是先天性缺陷的主要病毒原因。我们的小组和其他人表明，HCMV和其他疱疹病毒编码多个microRNA（miRNA），它们是小的21-24碱基对（BP）单链RNA物种，它们通过转录后机制调节基因表达。疱疹病毒miRNA已被证明针对许多不同的细胞和病毒过程，涉及病毒免疫识别，凋亡，细胞周期调节以及病毒潜伏期和裂解复制。在上一个资金期间，我们的小组已取得了重大进步，阐明了HCMV miRNA的功能作用。我们已经证明，HCMV miRNA之一有效地靶向并降低表达IE72蛋白，该蛋白显着降低了病毒复制，这表明了维持潜在病毒的作用。其次，我们还发现，HCMV miRNA协同合作以有效地下调单个基因，这可能解释了为什么病毒基因组中的单个miRNA敲除未显示表型效应。此外，HCMV miRNA似乎还靶向相同的细胞途径中的多个单独的细胞基因，包括两个潜在的抗病毒细胞因子 IL-1和TNF-以及NF？B抑制剂IKBA。最后，我们观察到双HCMV miRNA突变会导致病毒重新激活增加，而另外两个病毒miRNA双重突变体在体外的人类祖细胞（HPC）培养系统中未能重新激活。有趣的是，双突变剂中未能重新激活目标的HCMV miRNA之一，这两个NF？B激活剂表明已删除了NFKB激活以重新激活病毒。因此，在当前的建议中，我们将充分表征由HCMV miRNA靶向的IL-1和TNF-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-信号传导途径及其在CD34+ HPC中对病毒潜伏期和复制的功能相关性。我们还将检查新开发的人CD34+升级的NOD-SCIDIL2RGC NULL小鼠模型在潜伏期和重新激活过程中IL-1和TNF-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-tnf-oomna靶向的功能。我们假设病毒复制的HCMV miRNA复制与通过IL-1和TNF-途径的NFKB激活有关。