Project 5 - EBV Drivers of Oncogenesis and Novel Therapies

项目 5 - 肿瘤发生的 EBV 驱动因素和新疗法

• 批准号: 10910339
• 负责人: Shannon Celeste Kenney
• 金额: $ 7.24万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10910339
• 关键词: Affect; Attenuated; B-Lymphocytes; BCL2L11 gene; Bioinformatics; Burkitt Lymphoma; CDK4 gene; Cell Proliferation; Collaborations; Complex; Development; EZH2 gene; Epigenetic Process; Epstein-Barr Virus Infections; Epstein-Barr Virus latency; Generations; Genes; Genome; Growth; Helper-Inducer T-Lymphocyte; Hodgkin Disease; Human; Human Genetics; Human Herpesvirus 4; Immune response; Immunohistochemistry; Impairment; In Vitro; LMP1; Latent virus infection phase; Lymphocyte; Lymphoma; Lymphomagenesis; Lytic; Mediating; Methylation; MicroRNAs; Modeling; Molecular Biology; Molecular Genetics; Mutation; Oncogenes; Oncogenic Viruses; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Proteins; RNA; Role; Supporting Cell; T-Lymphocyte; TNFSF5 gene; Therapeutic; Tumor Suppressor Proteins; Umbilical Cord Blood; Viral; Viral Antigens; Viral Pathogenesis; Virus; Virus Latency; Xenograft Model; humanized mouse; immunogenicity; in vivo; inhibitor; insight; large cell Diffuse non-Hodgkin' s lymphoma; lymphoblastoid cell line; mouse model; mutant; novel therapeutic intervention; novel therapeutics; promoter; transcription factor; transforming virus; tumor; tumorigenesis

PROJECT 5 – PROJECT SUMMARY/ABSTRACT Epstein-Barr virus (EBV) latent infection of B lymphocytes in vitro results in their growth transformation; however, in vivo, the growth of EBV-infected lymphocytes is normally constrained by robust immune responses against viral antigens. As a result, EBV genes essential for growth in vitro (such as LMP1 and EBNA2) are often not expressed in EBV-positive lymphomas such as Burkitt lymphoma (BL), Hodgkin Disease and diffuse large B cell lymphomas. Thus, in vitro transformation studies cannot adequately model how EBV infection promotes common types of EBV-positive human lymphomas that have more stringent forms of viral latency. The EBV BARTs microRNAs, and EBNA3A, are amongst the very few EBV-encoded genes/microRNAs expressed in human BLs, and are likely to play important “driver” roles in this type of lymphoma. We have shown that BARTs play an important role in maintaining the viability of EBV-positive BLs in vitro, and in decreasing the immunogenicity of EBV-transformed lymphoblastoid cell lines. The latent EBNA3A protein encodes a transcription factor that is essential for in vitro growth of EBV-transformed B cells, and is thought to collaborate with the closely related EBNA3C protein to inhibit expression of important tumor suppressors (including p16, p15 and BIM) by inducing EZH2-mediated H3K27 trimethylation of their promoters. However, the roles of BARTs and EBNA3A in promoting EBV-induced lymphomas in vivo have not been well studied, particularly in the context of lymphomas with more stringent latency. EBV-infected humanized mice provide sophisticated models for understanding the complex interactions between EBV, T cells, cellular pathway alterations and the microenvironment. We have recently developed a new cord blood- humanized mouse model that allows EBV mutants that are non-transforming in vitro (including EBNA2- deleted EBV) to form lymphomas with stringent viral latency in vivo. We propose to use two different humanized mouse models to examine the roles of BARTs and EBNA3A for EBV-induced lymphomas in vivo, and to determine if drugs which block essential EBNA3A functions inhibit lymphoma development. In Aim 1, we will examine how loss of BARTs affects viral pathogenesis in the context of type III versus Wp-restricted viral latency, and examine potential mechanisms by which BARTs expression is upregulated in vivo. In Aim 2, we will use the cord blood-humanized mouse model to explore the role of the EBNA3A protein in vivo. In Aim 3, we will explore the therapeutic potential of drugs (CK4/6 and EZH2 inhibitors) that block essential EBNA3A/3C-regulated pathways. This project interacts extensively with Projects 3 and 4, and uses the cores for immunohistochemistry, bioinformatics, and generation of EBV mutant genomes. The results of these studies should provide key insights into the mechanism(s) by which stringent EBV infection causes lymphomas in vivo, and may identify new therapeutic approaches for treating EBV-induced lymphomas.

项目5 - 项目摘要/摘要 BARR病毒（EBV）在体外的B淋巴细胞潜在感染会导致其生长转化。 然而，在体内，EBV感染的淋巴细胞的生长通常受到强大的免疫的约束 对病毒抗原的反应。结果，EBV基因对于体外生长至关重要（例如LMP1和 EBNA2）通常在EBV阳性淋巴瘤（例如伯基特淋巴瘤（BL），霍奇金）中不表达 疾病和弥漫性大B细胞淋巴瘤。那是体外转化研究无法充分建模 EBV感染如何促进具有更严格的EBV阳性人类淋巴瘤的常见类型 病毒潜伏期的形式。 EBV BARTS microRNA和EBNA3A是极少数EBV编码的 在人类BLS中表达的基因/microRNA，可能在这种类型的中扮演重要的“驱动器”角色 淋巴瘤。我们已经表明，巴特在维持EBV阳性的生存力中起着重要作用 BLS体外，并降低EBV转化的淋巴细胞细胞系的免疫原性。潜在 EBNA3A蛋白编码一个转录因子，该因子对于EBV转换B细胞的体外生长至关重要， 并被认为与密切相关的EBNA3C蛋白合作以抑制重要肿瘤的表达 通过诱导的EZH2介导的H3K27三甲基化的补充剂（包括P16，P15和BIM） 发起人。但是，巴特和eBNA3A在促进EBV诱导的淋巴瘤中的作用具有 研究不好，尤其是在淋巴瘤的情况下具有更严格的潜伏期。 EBV感染 人源化的小鼠提供了复杂的模型，以理解EBV之间的复杂相互作用 细胞，细胞途径改变和微环境。我们最近开发了一种新的脐带血 人源化的小鼠模型允许在体外不转化的EBV突变体（包括EBNA2- 删除EBV）以在体内形成严格的病毒潜伏期。我们建议使用两个不同的 人性化小鼠模型检查BARTS和EBNA3A在EBV诱导的体内淋巴瘤的作用， 并确定阻断必需EBNA3A的药物是否功能抑制淋巴瘤的发育。在AIM 1中， 我们将研究BART的损失如何在III型与WP限制的情况下影响病毒发病机理 病毒潜伏期和检验在体内更新巴特表达的潜在机制。目标 2，我们将使用脐带血的小鼠模型来探索EBNA3A蛋白在体内的作用。在 AIM 3，我们将探索阻止必不可少的药物（CK4/6和EZH2抑制剂）的治疗潜力 EBNA3A/3C调节的途径。该项目与项目3和4进行了广泛的互动，并使用核心 用于免疫组织化学，生物信息学和EBV突变基因组的产生。这些结果 研究应提供有关严格的EBV感染机制的关键见解 体内淋巴瘤，并可能鉴定出用于治疗EBV诱导的淋巴瘤的新治疗方法。