Investigation of the role of HDAC activity in regulation of HCMV replication in the salivary epithelium

HDAC 活性在调节唾液上皮 HCMV 复制中的作用的研究

• 批准号: 10739852
• 负责人: WILLIAM E MILLER
• 金额: $ 24.3万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739852
• 关键词: 3-Dimensional; Ablation; Adult; American; Antiviral Agents; Area; Biological Models; Cell Nucleus; Cell model; Cells; Clinical; Collaborations; Complement; Complex; Cytomegalovirus; Cytomegalovirus Infections; Data; Defect; Development; Disease; Epithelial Cells; Epithelium; Exhibits; Fetal Development; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genes; Glean; Glycoproteins; Goals; Growth; HDAC1 gene; HDAC3 gene; Histone Deacetylase; Histone Deacetylase Inhibitor; Horizontal Disease Transmission; Human; Immunocompromised Host; In Vitro; Individual; Infection; Intervention; Investigation; Leukocytes; Life; Liquid substance; Liver; Medical; Microcephaly; Modeling; Molecular; Movement; Neurofibromin 2; Neurologic; Nuclear; Olfactory Mucosa; Oral; Organ; Parotid Gland; Pathogenesis; Pathologic; Persons; Physiological; Physiology; Play; Population; Process; Property; Proteins; Public Health; Publishing; Regulation; Role; Saliva; Salivary; Salivary Glands; Science; Severities; Signal Transduction; Spleen; Study models; Submandibular gland; Testing; Tissues; Transplant Recipients; United States; Viral; Viral Proteins; Virus; Virus Replication; Work; design; experimental study; in vitro Model; inhibitor; innovation; insight; knock-down; lytic replication; mutant; novel; organ transplant rejection; overexpression; pharmacologic; prevent; receptor; salivary cell; trafficking; transmission process; viral rescue; viral transmission

The human cytomegalovirus (HCMV) is a significant public health concern in the United States. Most important are the effects of the virus on developing fetuses and immunocompromised individuals where it causes a variety of pathological conditions ranging in severity from mild to life-threatening. Since HCMV is present in a persistent or latent form in 50-90% of the world’s adult population, the identification of viral gene products and mechanisms that contribute to viral trafficking, persistence, and horizontal transmission is an intense and important area of investigation. We have recently generated and published an in vitro model for studying HCMV replication in primary salivary derived epithelial cells called “salispheres”. These salisphere cells express genes typical of salivary acinar epithelial cells, mimic the physiology of the salivary gland, and serve as a valuable model to understand the basic parameters and mechanisms underlying HCMV replication in the salivary epithelium. Our exciting preliminary data indicates that HCMV strains deficient for the pentamer glycoprotein complex can be rescued for infection of salivary cells using histone deacetylase (HDAC) inhibitors. Moreover, our preliminary data suggest that the pentamer may be required to induce nuclear mobilization of pp71, where it can interact with the HDACs, inhibit their activity, and facilitate lytic replication. We also present preliminary data indicating the viral encoded GPCRs (vGPCRs) are essential for replication in salivary cells and that pharmacological HDAC inhibition can similarly rescue the defect exhibited by vGPCR null viruses. Our data suggest novel roles for both HCMV pentamer and vGPCRs in facilitating lytic replication in salivary cells. Based on our preliminary data, we hypothesize that the HCMV pentamer, vGPCRs, and pp71 work in concert to facilitate efficient viral replication in salivary epithelial cells leading to amplification and spread of virus from the salivary gland into the saliva. The proposed studies are highly significant to cytomegalovirus pathogenesis as the salivary gland and its secretions play an important role in horizontal transmission of virus, yet little is known about the viral mechanisms that facilitate infection and replication within the salivary epithelium. In aim 1, we will test whether the HCMV pentamer complex induces mobilization of the pp71 tegument protein into the nucleus to facilitate HDAC inhibition. In aim 2, we will test whether knockdown of HDAC1 or HDAC3 expression or overexpression of pp71 can rescue pentamer null viruses for efficient replication in salivary cells. In aim 3, we will determine whether HCMV vGPCR signaling works in concert with pentamer and pp71 to drive HCMV lytic replication in the salivary epithelial cells. The innovative experiments proposed in this application will generate important insight into the molecular and physiological properties of HCMV involved in salivary epithelial cell infection. Defining the mechanisms underlying salivary gland replication and spread could ultimately lead to the development of unique antivirals designed to prevent cytomegalovirus transmission via saliva.

人类巨细胞病毒 (HCMV) 在美国是一个重要的公共卫生问题，最重要的是该病毒对胎儿和免疫功能低下的个体的影响，导致多种病理状况，严重程度从轻微到危及生命。 HCMV 以持续或潜伏形式存在于世界上 50-90% 的成年人口中，识别有助于病毒贩运、持续存在和水平传播的病毒基因产物和机制是一个激烈而重要的领域。我们最近建立并发表了一个体外模型，用于研究原代唾液来源的上皮细胞（称为“唾液球”）中的 HCMV 复制，这些唾液球细胞表达唾液腺上皮细胞的典型基因，模仿唾液腺的生理学，并作为这是了解唾液上皮细胞中 HCMV 复制的基本参数和机制的一个有价值的模型。我们令人兴奋的初步数据表明，缺乏五聚体糖蛋白复合物的 HCMV 菌株可能是潜在的。此外，我们的初步数据表明，五聚体可能需要诱导 pp71 的核动员，从而与 HDAC 相互作用，抑制其活性并促进裂解复制。我们还提供了初步数据，表明病毒编码的 GPCR (vGPCR) 对于唾液细胞中的复制至关重要，并且药理学 HDAC 抑制同样可以挽救 vGPCR 无效病毒显示的缺陷。表明 HCMV 五聚体和 vGPCR 在促进唾液细胞中裂解性复制方面具有新作用。根据我们的初步数据，我们发现 HCMV 五聚体、vGPCR 和 pp71 协同作用，促进唾液上皮细胞中病毒的有效复制，从而导致扩增和复制。病毒从唾液腺传播到唾液中的研究对于巨细胞病毒的发病机制具有重要意义，因为唾液腺及其分泌物在巨细胞病毒的水平传播中发挥着重要作用。病毒，但对于促进唾液上皮内感染和复制的病毒机制知之甚少。在目标 1 中，我们将测试 HCMV 五聚体复合物是否诱导 pp71 外皮蛋白动员到细胞核中以促进 HDAC 抑制。 ，我们将测试 HDAC1 或 HDAC3 表达的敲低或 pp71 的过度表达是否可以挽救五聚体无效病毒在唾液细胞中的有效复制。在目标 3 中，我们将确定是否可以。 HCMV vGPCR 信号与五聚体和 pp71 协同作用，驱动唾液上皮细胞中的 HCMV 裂解性复制，本申请中提出的创新实验将为了解唾液上皮细胞感染中涉及的 HCMV 的分子和生理特性提供重要的见解。潜在的唾液腺复制和传播可能最终导致独特的抗病毒药物的开发，旨在防止巨细胞病毒通过唾液传播。