HCMV miRNA Regulation of Secretion and Formation of the Viral Assembly Compartment

HCMV miRNA 对病毒装配室的分泌和形成的调节

基本信息

批准号：
9250666
负责人：
William Jarvis Britt
金额：
$ 77.07万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9250666
关键词：
3&apos Untranslated Regions Affect Apoptosis Binding Biochemical Bioinformatics CDC42 gene Carrier Proteins Cell Cycle Progression Cell Death Cell Surface Proteins Cell physiology Cells Cellular Membrane Cytomegalovirus Cytoplasmic Protein Cytoskeleton Disease Endothelial Cells Engineering Environment Fibroblasts Gene Expression Gene Targeting Genes Genetic Transcription Goals Herpesviridae Homeostasis Human Immune Individual Infection Inflammatory Informatics Intracellular Membranes Investigation Lead Membrane Messenger RNA Metabolism Methodology MicroRNAs Morphogenesis Mutation Nucleic Acids Outcome Pathway interactions Phenotype Primary Infection Process Production Proteins RNA-Induced Silencing Complex Recombinants Recycling Regulation Repression Role SNAP23 gene Signal Transduction Small Interfering RNA Small RNA Transfection Translations Untranslated RNA Validation Viral Viral Proteins Virion Virus Virus Assembly Virus Diseases Virus Latency Virus Replication cell growth regulation cellubrevin cellular targeting cytokine human disease insight mutant novel protein transport public health relevance response small hairpin RNA trafficking

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) infection of primary cells results in massive alterations in normal cellular metabolism and regulation of cellular pathways that govern cell death, nucleic acid replication, cellular membrane homeostasis, cytoskeleton integrity, and intracellular trafficking of proteins. Numerous studies have documented the widespread dysregulation of cellular gene transcription following HCMV infection arguing that major changes in cellular phenotypes are controlled at a transcriptional level. However, crude regulation of essential and highly coordinated cellular functions such as protein transport and membrane homeostasis could result in a cellular environment that would also limit the production of infectious virus. Our recent studies have demonstrated that HCMV miRNAs regulate the secretory/endocytic pathway during infection by targeting genes encoding-key components of these pathways, thus providing another layer of HCMV regulation of cellular functions. Regulation of these pathways by HCMV miRNA results in inhibition of the release of pro-inflammatory cytokines and the reorganization of intracellular membranes to facilitate efficient production of infectious virus. In this project, the specific genes in the secretory/endocytic pathway targeted by HCMV miRNAs will be identified by a comprehensive approach combining PAR-CLIP methodology with informatics followed by rigorous validation. Initially, the role of validated miRNA targets in the regulation of trafficking in the endocytic an secretory pathways will be defined, followed by investigation of the role of these miRNA targets in cytokine trafficking and secretion, membrane reorganization, and infectious virus assembly. Validation of the role of these cellular targets of HCMV miRNAs in regulation of the secretory/endocytic pathways of the infected cells and their phenotypic effects on cytokine secretion and virus assembly will be accomplished by engineering recombinant HCMVs expressing specific combinations of shRNAs targeting genes within these pathways. It is anticipated that these studies will elucidate the role of these regulatory RNAs in the outcome of HCMV infection of the cell as well as defining a new and novel mode of regulation of essential cellular pathways leading to a cellular environment optimized for assembly of infectious virions.

描述（由申请人提供）：原代细胞的人巨细胞病毒（HCMV）感染导致正常细胞代谢和细胞途径调节的巨大改变，这些途径控制细胞死亡、核酸复制、细胞膜稳态、细胞骨架完整性和细胞内蛋白质运输许多研究记录了 HCMV 感染后细胞基因转录的广泛失调，认为细胞表型的主要变化是在转录水平上控制的。我们最近的研究表明，HCMV miRNA 通过靶向编码这些途径的关键成分的基因来调节感染过程中的分泌/内吞途径，因此蛋白质运输和膜稳态可能会导致细胞环境受到限制。 HCMV miRNA 对这些途径的调节导致促炎细胞因子的释放和细胞内膜的重组，以促进感染性病毒的有效产生。 HCMV miRNA 靶向的分泌/内吞途径中的基因将通过将 PAR-CLIP 方法与信息学相结合的综合方法进行鉴定，然后进行严格验证。定义，然后研究这些 miRNA 靶标在细胞因子运输和分泌、膜重组和感染性病毒组装中的作用，验证 HCMV miRNA 的这些细胞靶标在调节细胞因子中的作用。受感染细胞的分泌/内吞途径及其对细胞因子分泌和病毒组装的表型影响将通过工程重组 HCMV 来实现，这些重组 HCMV 表达这些途径中靶向基因的 shRNA 的特定组合。预计这些研究将阐明这些调节 RNA 的作用。细胞的 HCMV 感染的结果，以及定义一种新的、新颖的基本细胞途径调节模式，从而产生针对感染性病毒粒子组装而优化的细胞环境。