Defining mechanisms of KSHV pathogenesis using MHV68-KSHV chimeric viruses

使用 MHV68-KSHV 嵌合病毒定义 KSHV 发病机制

基本信息

批准号：
10243300
负责人：
James Craig Forrest
金额：
$ 52.58万
依托单位：
UNIV OF ARKANSAS FOR MED SCIS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-14 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10243300
关键词：
Acquired Immunodeficiency Syndrome Animal Model Antiviral Agents Biological Models Cell Lineage Cell physiology Cells Chimera organism Chronic Cytometry DNA Tumor Viruses DNA-Binding Proteins Development Disease Episome Evaluation Fostering G-Protein-Coupled Receptors Gene Transfer Genes Genetic Transcription Genome Goals HIV Herpesviridae Infections Human Human Herpesvirus 8 Immune Immune response Immunity Individual Infection Inflammation Inflammatory Kaposi Sarcoma Knock-in Laboratory mice Lymphocyte Lytic Maintenance Malignant Neoplasms Methods Modeling Molecular Morbidity - disease rate Multicentric Angiofollicular Lymphoid Hyperplasia Mus Oncogenic Oncoproteins Organism Outcome Pathogenesis Pathogenicity Persons Preclinical Testing Prevention Process Production Proteins Risk Rodent Role Signal Pathway System Time Trans-Activators Transcriptional Regulation Transgenic Mice Viral Viral Genes Viral Pathogenesis Viral Proteins Virus Virus Diseases Virus Latency Virus Replication Work adaptive immune response adaptive immunity antiretroviral therapy cancer risk cell type chronic infection experimental study gammaherpesvirus gene replacement humanized mouse improved in vivo infected B cell latency-associated nuclear antigen lytic replication mortality next generation novel strategies pathogen prevent primary effusion lymphoma promoter protein function response single-cell RNA sequencing targeted treatment therapy development tissue culture tool viral cyclin

项目摘要

PROJECT SUMMARY Gammaherpesviruses (GHVs) are DNA tumor viruses that establish lifelong, chronic infections of host lymphocytes. The expression of viral gene products that manipulate host cell physiology and thwart antiviral immune responses places the infected host at risk for numerous cancers. For individuals with AIDS, infection by Kaposi sarcoma-associated herpesvirus (KSHV) is a major cause of morbidity and mortality. However, KSHV does not readily infect mice, which complicates attempts to define mechanisms by which KSHV establishes long- term infections and disease. To overcome this barrier, we have used a chimeric virus approach in which KSHV genes are transferred into the closely related virus, murine gammaherpesvirus 68 (MHV68), which is a natural rodent pathogen that readily infects laboratory mice. We used this MHV68-KSHV chimeric virus approach to evaluate the KSHV latency-associated nuclear antigen (kLANA) during productive viral replication, latency establishment, and maintenance. We found that kLANA was sufficient to replace MHV68 LANA (mLANA) for viral latency. However, kLANA, but not mLANA, suppressed MHV68 lytic replication by inhibiting the activity of the promoter for lytic transactivator RTA. Our work demonstrates that chimeric viruses offer opportunities for dissecting functions of KSHV proteins in a living host, and that this system can define evolutionarily acquired features of a viral oncoprotein. Experiments described in this proposal will further define functions of KSHV LANA in viral pathogenesis and evolutionary divergence from mLANA. We will use the repressed lytic replication of the kLANA-expressing chimeric virus to define how the amplitude of lytic replication influences immunity to infection and oncogenic potential of the virus. Finally, we will extend our chimeric virus studies with LANA, dovetailing them with MHV68-KSHV chimeric viruses for v-Cyclin and v-GPCR, to develop the next generation of chimeras in which multiple MHV68 genes are replaced with their KSHV counterparts. Through this work we will improve small animal models to enable molecular mechanistic studies of KSHV oncoproteins in viral infection and disease and provide a preclinical testing platform to foster the development of therapies that target LANA and other viral proteins to treat or prevent KSHV-related cancers.

项目概要伽马疱疹病毒 (GHV) 是一种 DNA 肿瘤病毒，可引起宿主的终生慢性感染淋巴细胞。操纵宿主细胞生理学并阻止抗病毒药物的病毒基因产物的表达免疫反应使受感染的宿主面临多种癌症的风险。对于艾滋病患者来说，感染卡波西肉瘤相关疱疹病毒（KSHV）是发病和死亡的主要原因。然而，KSHV 不易感染小鼠，这使得定义 KSHV 建立长期传播机制的尝试变得复杂术语感染和疾病。为了克服这一障碍，我们使用了嵌合病毒方法，其中 KSHV 基因被转移到密切相关的病毒鼠伽马疱疹病毒 68 (MHV68) 中，这是一种天然的病毒。容易感染实验室小鼠的啮齿动物病原体。我们使用这种 MHV68-KSHV 嵌合病毒方法评估生产性病毒复制期间的 KSHV 潜伏期相关核抗原 (kLANA)、潜伏期建立、维护。我们发现 kLANA 足以替代 MHV68 LANA (mLANA) 病毒潜伏期。然而，kLANA（而非 mLANA）通过抑制 MHV68 的活性来抑制 MHV68 裂解性复制。裂解反式激活子 RTA 的启动子。我们的工作表明嵌合病毒提供了机会剖析 KSHV 蛋白在活体宿主中的功能，并且该系统可以定义进化获得的病毒癌蛋白的特征。本提案中描述的实验将进一步定义 KSHV LANA 的功能病毒发病机制和与 mLANA 的进化分歧。我们将使用抑制的裂解复制表达 kLANA 的嵌合病毒，用于定义裂解复制的幅度如何影响对感染的免疫力和病毒的致癌潜力。最后，我们将扩展我们与 LANA 的嵌合病毒研究，吻合将它们与用于 v-Cyclin 和 v-GPCR 的 MHV68-KSHV 嵌合病毒一起开发下一代嵌合体其中多个 MHV68 基因被替换为相应的 KSHV 基因。通过这项工作我们将改进小动物模型可用于病毒感染和疾病中 KSHV 癌蛋白的分子机制研究并提供临床前测试平台，以促进针对 LANA 和其他病毒的疗法的开发治疗或预防 KSHV 相关癌症的蛋白质。