HCMV regulation of monocyte/macrophage host cell signaling in viral reactivation

HCMV 对病毒再激活中单核细胞/巨噬细胞宿主细胞信号传导的调节

• 批准号: 10327952
• 负责人: ANDREW D YUROCHKO
• 金额: $ 39.57万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10327952
• 关键词: Address; Animal Model; Biological; Biology; CD34 gene; Cell Differentiation process; Cell model; Cells; Chemicals; Critical Pathways; Cytomegalovirus; Data; Disease; Environment; Epidermal Growth Factor Receptor; Event; Failure; Funding; Gene Mutation; Genetic Transcription; Goals; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Infection; Knowledge; Laboratories; Link; MicroRNAs; Modification; Molecular Profiling; Morbidity - disease rate; Multiomic Data; Myelogenous; Myeloid Cells; Organ Transplantation; Pathway interactions; Phenotype; Play; Prevention; Process; Regulation; Role; SRC gene; Signal Pathway; Signal Transduction; Site; Solid; Testing; Time; Tissues; Translating; Transplant Recipients; Viral; Viral Genes; Virion; Virus; Virus Latency; Virus Replication; Work; arm; cell type; clinically relevant; design; directed differentiation; experimental study; gene product; humanized mouse; immune checkpoint blockade; improved outcome; in vivo; inhibitor; insight; knock-down; latent infection; macrophage; macrophage product; monocyte; mortality; mouse model; multiple omics; mutant; novel; pathogen; prevent; programs; reactivation from latency; response; small hairpin RNA; therapeutic target

SUMMARY – PROJECT 5 Human cytomegalovirus (HCMV) is a significant cause of morbidity and mortality after hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT) due to reactivation or a new infection in these transplant recipients. Our combined PPG laboratories have shown that HCMV infection of CD34+ hematopoietic progenitor cells (HPCs) the site of HCMV latency and subsequent reactivation alters hematopoietic events to favor HPC differentiation towards the myeloid lineage. To fully understand HCMV reactivation process, we need not only understand the process of how reactivating virus in CD34+ HPC directs differentiation towards the myeloid lineage, but also mechanistically how HCMV reactivation in monocytes directs signaling to promote differentiation towards productive macrophages. Through our collaborative program, we have identified viral genes (e.g., UL135, US28, UL7/8) and miRNAs (e.g. miR-US22) that are required for reactivation. Furthermore, through the use of deletion mutant viruses that fail to reactivate, we have identified the existence of blockades to HCMV reactivation that are associated with differentiation into macrophages. The signaling events important for reactivation of latent virus in tissue macrophages remains poorly understood. We suggest temporally unique roles for UL135, US28 and UL7/8 and a unique intersection of these gene products with HCMV miRNAs expressed during latency in usurping the signaling necessary to promote reactivation in this essential cell type. We hypothesize distinct signaling events in monocyte to macrophage differentiation relieve blockades to reactivation and these reactivation events are specifically driven by modifications to cellular signaling by the combined effort of these viral factors. To test our hypothesis, we propose the following aims. Specific Aim 1. Defining signaling networks for HCMV latency and reactivation in monocytes and macrophages. We hypothesize that functionally distinct host cell responses in monocyte-to-macrophage differentiation are essential for reactivation. Specific Aim 2. Determine checkpoint blockades for viruses that fail to reactivate. We hypothesize that a failure in specific cellular signaling reprogramming during reactivation will result in a cell that phenotypically is unable to support viral replication. Specific Aim 3. Define the signaling nodes modulated by infection for reactivation in monocytes/macrophages. We hypothesize that the EGFR and RhoA signaling pathways are critical for HCMV latency and reactivation in macrophages. IMPACT: These proposed aims will allow us to gain greater insight and as we integrate our collective work, we hope to translate the results into novel molecular signatures designed to improve the outcome of an infection that remains a significant problem in transplant recipients.

摘要 - 项目5 人类巨细胞病毒（HCMV）是造血后发病率和死亡率的重要原因 由于重新激活或新的感染，干细胞移植（HSCT）和固体器官移植（SOT） 移植接受者。我们组合的PPG实验室表明，CD34+造血的HCMV感染 祖细胞（HPC）HCMV潜伏期的部位和随后的重新激活改变造血事件 偏向于HPC分化髓样谱系。为了完全了解HCMV重新激活过程，我们需要 不仅了解CD34+ HPC中如何重新激活病毒的过程 髓样谱系，也机械地，单核细胞中的HCMV重新激活如何将信号引导到 促进对生产巨噬细胞的分化。 通过我们的协作计划，我们确定了病毒基因（例如UL135，US28，UL7/8）和 重新激活所需的miRNA（例如miR-us22）。此外，通过使用缺失突变体 未能重新激活的病毒，我们已经确定了HCMV重新激活的封锁存在 与分化为巨噬细胞有关。信号事件对于潜在病毒重新激活很重要 在组织中，巨噬细胞的理解仍然很差。我们建议使用UL135，US28暂时独特的角色 和UL7/8以及这些基因产物与HCMV miRNA的独特交集。 在这种基本细胞类型中促进重新激活所必需的信号时的延迟。我们 假设单核细胞中的不同信号事件对巨噬细胞分化的救援阻塞至重新激活 这些重新激活事件是由通过合并努力对细胞信号的修改特别驱动的 这些病毒因素。为了检验我们的假设，我们提出以下目的。特定目标1。定义信号 单核细胞和巨噬细胞中HCMV潜伏期和重新激活的网络。我们在功能上假设这一点 单核细胞对摩噬细胞分化的不同宿主细胞反应对于重新激活至关重要。具体的 AIM 2。确定未重新激活病毒的检查点阻滞。我们假设特定的失败 重新激活过程中的细胞信号传导重新编程将导致表型无法支撑的细胞 病毒复制。特定目的3。定义通过感染调节的信号节点以重新激活 单核细胞/巨噬细胞。我们假设EGFR和RhoA信号通路对于HCMV至关重要 巨噬细胞的潜伏期和重新激活。 影响：这些提议的目标将使我们能够获得更大的见识，并且随着我们整合我们的集体 工作，我们希望将结果转化为新的分子特征，旨在改善 在移植受者中仍然是一个重大问题的感染。