Targeting Nuclear HSF1 as a Novel Anti-HCMV Strategy

靶向核 HSF1 作为一种新型抗 HCMV 策略

基本信息

批准号：
10656697
负责人：
Gary Ching Tao Chan
金额：
$ 64.9万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-10 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10656697
关键词：
Ablation Adverse effects Advocate Animal Model Antiviral Agents Antiviral Therapy Attenuated Biological Bone Marrow Cells Cells Cessation of life Chronic Clinical Cytomegalovirus Cytomegalovirus Infections Data Development Disease Distal Drug Kinetics Drug toxicity Failure Fibroblasts Foundations Future Genes Genetic Heat-Shock Response Human Immunocompromised Host Infection Infiltration Inflammatory Kinetics Lytic Macrophage Mediating Monitor Multiple Organ Failure Mus Myeloid Cells Nuclear Opportunistic Infections Organ Organ Transplantation Pathogenesis Peripheral Persons Pharmaceutical Preparations Pharmacodynamics Pharmacology Prophylactic treatment Proteins Regimen Resistance Role Site Skin Stress Testing Therapeutic Tissues Transplant Recipients Transplantation Viral Viral Genome Viral Physiology Virus Virus Replication antagonist biological adaptation to stress cell type design drug discovery drug efficacy efficacy evaluation genetic approach heat-shock factor 1 high risk human model human tissue immunosuppressed improved in vivo in vivo evaluation inflammatory milieu inhibitor insight knock-down latent infection lytic replication monocyte mouse model neonate new therapeutic target novel organ transplant recipient permissiveness pharmacologic prevent standard of care tool transcription factor transcriptome transcriptome sequencing translatome transplant model viral rebound

项目摘要

Project Summary Human cytomegalovirus (HCMV) infects the majority of people in the world and can cause serious disease in immunocompromised patients and neonates. The virus establishes life-long latency in bone marrow cells and disseminates to peripheral organs in quiescently infected monocytes. Antiviral therapy delays virus replication, but does not eliminate infected cells. Virus rebound, resistance, and drug toxicity complicate treatment and create a strong demand for improved therapeutics. We advocate that the suppression of HCMV replication must be in combination with the killing of infected monocytes. We found that HCMV infection of fibroblasts and monocytes rapidly stimulated the activity of heat shock factor (HSF) 1, a stress-responsive transcription factor, in a distinct fashion from canonical activation induced by heat shock (HS). Using a novel tool compound called DTHIB, which has been validated to selectively inhibit HSF1 activity, we found inhibition of HSF1 with DTHIB attenuated HCMV lytic replication and stimulated death of latently infected monocytes. These studies provide the beginnings of a proof-of-concept study that HSF1 antagonists may have the capacity to provide the double “hit” necessary to suppress HCMV replication and eliminate latently infected myeloid cells in a single drug. Thus, our central hypothesis is that inhibition of HSF1 with the tool compound DTHIB will limit both infection and spread within an infected host by concomitantly attenuating HCMV lytic replication in permissive cell types and eliminating latently infected monocytes. The first aim will continue to evaluate the antiviral potential of DTHIB as an inhibitor of HCMV lytic replication by examining the drug efficacy on different HCMV permissive cell types, viral strains, and multiplicities of infection (MOIs). We will also conduct transcriptome (RNA sequencing) analyses and functional studies using DTHIB to identify genes dependent on HCMV-induced HSF1 activity responsible for promoting lytic replication and the impact of DTHIB on the expression of this HCMV- induced, HSF1-dependent gene profile. The second aim will continue to assess the ability of DTHIB to stimulate the death of latently infected monocytes by testing the selective drug toxicity on monocytes infected with different viral strains and at different MOIs. In conjunction, we will perform translatome (polysomal profiling) analyses and functional studies using DTHIB to identify HSF1-dependent genes responsible for promoting the survival of latently infected monocytes. The third aim will assess the in vivo antiviral activity of DTHIB on lytic replication, viral spread, and pathogenesis using a novel murine transplant model with human skin organ, which can simultaneously monitor HCMV replication in human tissue as well as monitor monocyte-mediated HCMV spread to distal sites.

项目概要人类巨细胞病毒 (HCMV) 感染世界上大多数人，并可在以下国家引起严重疾病：该病毒在免疫功能低下的患者和新生儿中建立了终生潜伏期。在静止感染的单核细胞中传播到外周器官，抗病毒治疗会延迟病毒复制，但不能消除受感染的细胞，病毒反弹、耐药性和药物毒性使治疗复杂化。我们主张必须抑制 HCMV 复制。我们发现HCMV感染与成纤维细胞的杀伤相结合。单核细胞迅速刺激热休克因子（HSF）1（一种应激反应转录因子）的活性，以一种不同于热休克（HS）诱导的典型激活的方式使用一种称为新型工具化合物。 DTHIB，已被验证可以选择性抑制 HSF1 活性，我们发现 DTHIB 对 HSF1 具有抑制作用这些研究提供了减弱 HCMV 裂解性复制并刺激潜伏感染单核细胞死亡的方法。概念验证研究的开始，HSF1 拮抗剂可能具有提供双重作用的能力单一药物中抑制 HCMV 复制和消除潜伏感染的骨髓细胞所必需的“打击”。我们的中心假设是用工具化合物 DTHIB 抑制 HSF1 将限制两种感染并通过同时减弱 HCMV 裂解性复制而在受感染宿主内传播细胞类型并消除潜伏感染的单核细胞。首要目标将继续评估抗病毒药物。通过检查不同 HCMV 的药物功效，探讨 DTHIB 作为 HCMV 裂解性复制抑制剂的潜力允许的细胞类型、病毒株和感染复数 (MOIs) 我们还将进行转录组 (RNA) 分析。测序）使用 DTHIB 进行分析和功能研究来鉴定依赖于 HCMV 诱导的 HSF1 的基因负责促进裂解复制的活性以及 DTHIB 对该 HCMV- 表达的影响第二个目标将继续评估 DTHIB 的刺激能力。通过测试对感染不同病毒的单核细胞的选择性药物毒性，观察潜伏感染单核细胞的死亡情况病毒株和不同的 MOI 结合起来，我们将进行翻译组（多核糖体分析）分析和使用 DTHIB 的功能研究来鉴定负责促进存活的 HSF1 依赖性基因第三个目标是评估 DTHIB 对裂解复制的体内抗病毒活性，使用具有人类皮肤器官的新型小鼠移植模型来研究病毒传播和发病机制，该模型可以同时监测人体组织中的 HCMV 复制以及单核细胞介导的 HCMV 传播到远端部位。