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.

项目摘要 γ掌病毒（GHVS）是DNA肿瘤病毒，可建立终生的宿主慢性感染淋巴细胞。操纵宿主细胞生理和阻止抗病毒的病毒基因产物的表达免疫反应使受感染的宿主面临许多癌症的风险。对于有艾滋病的人，感染 Kaposi肉瘤相关的疱疹病毒（KSHV）是发病率和死亡率的主要原因。但是，KSHV 不容易感染小鼠，这使尝试定义KSHV建立长期的机制的尝试复杂化期感染和疾病。为了克服这一障碍，我们使用了KSHV的嵌合病毒方法将基因转移到密切相关的病毒中，鼠γ瘤病毒68（MHV68），这是自然的容易感染实验室小鼠的啮齿动物病原体。我们使用了这种MHV68-KSHV嵌合病毒方法在生产性病毒复制期间评估KSHV潜伏期相关的核抗原（Klana）建立和维护。我们发现Klana足以代替MHV68 LANA（MLANA）病毒潜伏期。但是，克拉纳（Klana），但不是姆拉纳（Mlana 裂解反式激活器RTA的启动子。我们的工作表明，嵌合病毒为在活宿主中剖析KSHV蛋白的功能，并且该系统可以定义进化获取病毒癌蛋白的特征。该提案中描述的实验将进一步定义KSHV LANA的功能在病毒发病机理和与麦拉纳的进化差异中。我们将使用压抑的裂解复制表达克拉纳的嵌合病毒，以定义裂解复制的幅度如何影响感染的免疫力和病毒的致癌潜力。最后，我们将使用拉娜（Lana）扩展嵌合病毒研究它们使用V-Cyclin和V-GPCR的MHV68-KSHV嵌合病毒，以开发下一代的嵌合体其中多个MHV68基因被其KSHV对应物取代。通过这项工作，我们将改善小动物模型能够在病毒感染和疾病中对KSHV癌蛋白进行分子机理研究并提供一个临床前测试平台，以促进针对LANA和其他病毒的疗法的开发蛋白质治疗或预防KSHV相关的癌症。