HCMV US28 regulation of host cell signaling in viral latency and hematopoiesis

HCMV US28 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：
9980283
负责人：
DANIEL N STREBLOW
金额：
$ 25.61万
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/9980283
关键词：
Affect Allogenic BLT mice Binding Biosensor Bone Marrow Transplantation CCL2 gene CD34 gene CREB1 gene CX3C Chemokines Cell Differentiation process Cell Lineage Cell Maintenance Cell model Cells Characteristics Cues Cytomegalovirus Development Disease EGF Signaling Pathway EGF gene Ensure Epithelial Epithelium Equilibrium Event Fractalkine Gene Expression Genetic Transcription Goals Graft Rejection Growth Factor Receptors Hematopoiesis Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Herpesviridae Human Immediate-Early Genes Immune In Vitro Infection Inflammation Inflammatory Ligands Mediating Mediator of activation protein MicroRNAs Modeling Molecular Monomeric GTP-Binding Proteins Morbidity - disease rate Mutation Myelogenous Myelosuppression Organ Transplantation PTK2 gene Pathway interactions Population Proteins RANTES Receptor Signaling Recombinant Delta Chemokine Regulation Role Signal Pathway Signal Transduction Signaling Molecule Smooth Muscle Solid Stem cell transplant Stimulus Therapeutic Intervention Transcriptional Activation Transplant Recipients Transplant-Related Disorder Transplantation Viral Viral Proteins Virus Virus Latency Virus Replication base beta-Chemokines cell motility chemokine chemokine receptor gene product graft failure humanized mouse in vivo inhibitor/antagonist latency-associated protein macrophage migration monocyte mortality mouse model multiple omics novel therapeutics nuclear factors of activated T-cells peripheral blood prevent programs reactivation from latency receptor expression receptor-mediated signaling response targeted treatment transcriptome sequencing

项目摘要

PROJECT 3 SUMMARY Human cytomegalovirus (HCMV) is a -herpesvirus infecting 44-100% of the population and remains a significant cause of morbidity and mortality in solid organ transplant (SOT) and allogeneic hematopoietic stem cell transplant (SCT) recipients. Infection in SCT patients is often associated with myelosuppression and graft failure due to virus reactivation from latency, but the associated mechanisms are still largely unknown. HCMV encodes multiple latency-associated gene products including the chemokine receptor US28, which binds CC- chemokines as well as the CX3C-chmokine Fractalkine. US28 signals in both ligand-independent and ligand- dependent manners, but the downstream signaling pathways and consequences are unique for each. Importantly, herein we demonstrate that US28 is required for latency and/or reactivation in both in vitro CD34+ hematopoietic progenitor cells (HPCs) as well as in a humanized mouse model. In addition, we show that US28 is sufficient to promote CD34+ HPC differentiation into myeloid lineage cells. Therefore, based upon our exciting new findings, we hypothesize that US28 ligand-independent signaling during latency helps to maintain latency and that ligand-specific signaling promotes CD34+ HPC differentiation into a reactivation/replication competent myeloid lineage cell to ensure virus replication under appropriate conditions including inflammation. US28 signaling may also contribute to other aspects of latency by acting as a biosensor to promote migration towards inflammatory chemokines, and possibly by increasing immediate early gene expression during reactivation. Defining the role of US28 in latency and reactivation, determining how US28 signaling intersects with EGFR signaling, and understanding how US28 signaling influences other HCMV proteins (Projects 1 and 4) and miRNAs (Project 2) expressed during latency and manipulating EGFR signaling (Project 5) are the goals of this proposal. We propose the following specific aims: 1) To determine how US28 signaling in CD34+ HPCs intersects with EGFR signaling pathways using a multi-omics approach; 2) To determine what molecular characteristics of US28 mediate HCMV latency and reactivation using both in vitro CD34+ HPCs and in vivo humanized BLT mouse models; 3) To determine what US28 signaling pathways promote CD34+ HPC hematopoiesis also using both in vitro CD34+ HPC and in vivo huBLT models. Results of this study will generate new virus latency and reactivation paradigms promoting the development of novel therapies to prevent virus reactivation and to treat HCMV-mediated myelosuppression.

项目 3 摘要人类巨细胞病毒 (HCMV) 是一种 -疱疹病毒，感染 44-100% 的人口，并且仍然是一种实体器官移植（SOT）和异体造血干细胞发病率和死亡率的重要原因细胞移植 (SCT) 受者的感染通常与骨髓抑制和移植物相关。由于病毒潜伏期重新激活而导致失败，但相关机制仍不清楚。编码多种潜伏相关基因产物，包括趋化因子受体 US28，它结合 CC- 趋化因子以及配体非依赖性和配体依赖性的 CX3C 趋化因子 US28 信号。依赖方式，但下游信号通路和后果对于每种方式都是独特的。重要的是，在此我们证明 US28 是体外 CD34+ 中潜伏和/或重新激活所必需的此外，我们在造血祖细胞（HPC）以及人源化小鼠模型中证明了这一点。 US28 足以促进 CD34+ HPC 分化为骨髓谱系细胞，因此，根据我们的研究。令人兴奋的新发现，我们追求潜伏期 US28 配体独立信号传导有助于维持潜伏期和配体特异性信号传导促进 CD34+ HPC 分化为重新激活/复制有能力的骨髓谱系细胞确保病毒在包括炎症在内的适当条件下复制。 US28 信号传导还可能通过充当生物传感器来促进迁移，从而导致延迟的其他方面炎症趋化因子，并可能通过增加早期基因表达定义 US28 在延迟和重新激活中的作用，确定 US28 信号传导的交叉方式。 EGFR 信号传导，并了解 US28 信号传导如何影响其他 HCMV 蛋白（项目 1 和 4) 和在潜伏期表达并操纵 EGFR 信号传导 (项目 5) 的 miRNA (项目 2) 是我们提出以下具体目标： 1) 确定 CD34+ 中 US28 信号传导的方式。使用多组学方法 HPC 与 EGFR 信号通路相交叉 2) 确定何种分子；使用体外 CD34+ HPC 和体内 US28 的特征介导 HCMV 潜伏期和再激活人源化 BLT 小鼠模型；3) 确定哪些 US28 信号通路促进 CD34+ HPC 还使用体外 CD34+ HPC 和体内 huBLT 模型进行造血作用。产生新的病毒潜伏和重新激活范例，促进新疗法的开发预防病毒重新激活并治疗 HCMV 介导的骨髓抑制。