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

HCMV US28 在病毒潜伏期和再激活过程中对宿主细胞信号传导的调节

基本信息

批准号：
10327950
负责人：
DANIEL N STREBLOW
金额：
$ 40.04万
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 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10327950
关键词：
Affect Allogenic Binding Bone Marrow Transplantation C-terminal CCL2 gene CD34 gene CX3C Chemokines Cell Differentiation process Cell Lineage Cell Maintenance Cells Cercopithecine Herpesvirus 1 Collaborations Couples Cytomegalovirus Data Development Dissection Epidermal Growth Factor Receptor Fractalkine Genes Goals Graft Rejection Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human In Vitro Infection Ligand Binding Ligands Link MEKs Mediating MicroRNAs Molecular Morbidity - disease rate Mutation Mutation Analysis Myelogenous Myelosuppression N-terminal Organ Organ Transplantation Pathway interactions Play Point Mutation Population Process Proteins RANTES Receptor Signaling Recombinant Delta Chemokine Regulation Role Signal Pathway Signal Transduction Signaling Molecule Solid Stem cell transplant System Tail Time Transplant Recipients Undifferentiated Viral Viral Proteins Virus Virus Latency beta-Chemokines chemokine chemokine receptor functional outcomes gene product graft failure humanized mouse in vitro Model in vivo in vivo Model inhibitor knock-down member monocyte mortality mutant neutralizing antibody novel therapeutics prevent reactivation from latency rho sensor sensor technology

项目摘要

SUMMARY - PROJECT 3 Human cytomegalovirus (HCMV) is a b-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-chemokine Fractalkine. We have demonstrated that US28 ligand-dependent signaling is required to maintain viral latency in vitro in CD34+ hematopoietic progenitor cells (HPCs) and in vivo in humanized mice. US28 was required in both systems for reactivation of latent HCMV and promotes CD34+ HPC differentiation into myeloid lineage cells, indicating that US28 plays a dynamic role in both processes. While we have identified that ligand binding activity for US28 is required for HCMV latency, we do not yet know whether this effect is ligand-specific. In addition, we have recently demonstrated that HCMV-encoded chemokine ligands modulate US28 signaling. We hypothesize that both the host CC chemokines and viral chemokines UL146/UL147 promote latency by activating specific signaling pathways while other US28 ligands help direct reactivation by modifying signaling. We hypothesize that US28 ligand binding activity (ligand-specific) is required to maintain latency by interfacing with EGFR signaling and during reactivation US28 antagonizes EGFR signaling and shifts to activating RhoA pathways to enhance productive replication. In Specific Aim 1, we will determine which US28 ligands reprogram cells during latency and reactivation using neutralizing antibodies against host and viral chemokines, viral mutants that impact the ability of US28 to selectively bind ligands, and viral mutants lacking the viral chemokines. In SA2, we will utilize our US28 interactome data generated using the BirA-Turbo proximity sensor technology to identify signaling factors that play a role in latency and reactivation. Through mutational analyses we have identified specific regions of US28, in the C’terminal tail and intracellular loop 3 domains, that modify US28 signaling. We will determine what effect these mutations have on both the US28 interactome and latency and reactivation using in vitro and in vivo models. Lastly, in SA3, we will define how US28 signaling influences other HCMV proteins (Projects 1 and 4) and miRNAs (Project 2) with specific focus on how US28 intersects with the EGFR pathway, to contribute to the combined HCMV-manipulation of EGFR signaling (Project 5) and therefore control of latency and reactivation within the overarching goals of this proposal. Results of this study will generate new virus latency and reactivation paradigms promoting the development of novel therapies to prevent virus reactivation.

摘要 - 项目 3 人类巨细胞病毒 (HCMV) 是一种 b 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 CX3C 趋化因子 Fractalkine 我们已经证明 US28 配体依赖性。需要信号传导来维持体外 CD34+ 造血祖细胞 (HPC) 和体内的病毒潜伏期在人源化小鼠中，两个系统都需要 US28 来重新激活潜伏的 HCMV 并促进 CD34+。 HPC 分化为骨髓谱系细胞，表明 US28 在这两个过程中都发挥着动态作用。我们已经确定 US28 的配体结合活性是 HCMV 潜伏期所必需的，但我们还不知道是否这种效应是配体特异性的。此外，我们最近证明了 HCMV 编码的趋化因子配体。我们勇敢地指出宿主 CC 趋化因子和病毒趋化因子。 UL146/UL147 通过激活特定信号通路来促进潜伏期，而其他 US28 配体则有助于指导我们勇敢地说，US28 配体结合活性（配体特异性）是必需的。通过与 EGFR 信号传导连接来维持潜伏期，并且在重新激活期间 US28 拮抗 EGFR 信号传导并转向激活 RhoA 途径以增强高效复制。在具体目标 1 中，我们将确定。 US28 配体在潜伏期和重新激活期间使用针对宿主的中和抗体重新编程细胞和病毒趋化因子、影响 US28 选择性结合配体能力的病毒突变体和病毒突变体在 SA2 中，我们将利用 BirA-Turbo 生成的 US28 相互作用组数据。接近传感器技术可识别在延迟和重新激活中起作用的信号因素。突变分析我们已经确定了 US28 的特定区域，位于 C' 末端尾部和细胞内环 3 改变 US28 信号传导的结构域，我们将确定这些突变对 US28 有何影响。最后，在 SA3 中，我们将定义如何使用体外和体内模型进行相互作用组、潜伏期和重新激活。 US28 信号传导特别关注其他 HCMV 蛋白（项目 1 和 4）和 miRNA（项目 2）关于 US28 如何与 EGFR 通路交叉，以促进 EGFR 的组合 HCMV 操纵信令（项目 5），因此在该项目的总体目标内控制延迟和重新激活这项研究的结果将产生新的病毒潜伏和重新激活范例，促进开发新疗法以防止病毒重新激活。