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

基本信息

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
项目类别：
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&apos s lymphoma lymphoblastoid cell line mouse model mutant novel therapeutic intervention novel therapeutics promoter transcription factor transforming virus tumor tumorigenesis

项目摘要

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 EB病毒（EBV）在体外潜伏感染B淋巴细胞导致其生长转化；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系统的限制。因此，EBV 基因对体外生长至关重要（例如 LMP1 和 EBNA2）通常在 EBV 阳性淋巴瘤中不表达，例如伯基特淋巴瘤 (BL)、霍奇金淋巴瘤因此，体外转化研究无法充分模拟疾病和弥漫性大 B 细胞淋巴瘤。 EBV 感染如何促进常见类型 EBV 阳性人类淋巴瘤的发生，这些淋巴瘤具有更严格的要求 EBV BART microRNA 和 EBNA3A 是极少数 EBV 编码的病毒潜伏期形式。人类 BL 中表达的基因/microRNA 可能在此类疾病中发挥重要的“驱动”作用我们已经证明 BART 在维持 EBV 阳性的生存能力方面发挥着重要作用。 BLs 体外，并降低 EBV 转化的淋巴母细胞系的免疫原性。 EBNA3A 蛋白编码一种转录因子，该因子对于 EBV 转化的 B 细胞的体外生长至关重要，并被认为与密切相关的 EBNA3C 蛋白协同抑制重要肿瘤的表达通过诱导 EZH2 介导的 H3K27 三甲基化来抑制抑制因子（包括 p16、p15 和 BIM）然而，BART 和 EBNA3A 在体内促进 EBV 诱导的淋巴瘤中的作用已经被证实。尚未得到充分研究，特别是在潜伏期更严格的 EBV 感染的情况下。人源化小鼠为理解 EBV、T 之间复杂的相互作用提供了复杂的模型我们最近开发了一种新的脐带血- 人源化小鼠模型，允许体外非转化的 EBV 突变体（包括 EBNA2- 删除的 EBV）以在体内形成具有严格病毒潜伏期的淋巴瘤。人源化小鼠模型，用于检查 BART 和 EBNA3A 对 EBV 诱导的体内淋巴瘤的作用，并确定阻断 EBNA3A 基本功能的药物是否会抑制淋巴瘤的发展。我们将研究 BART 的缺失如何影响 III 型病毒与 Wp 限制型病毒的发病机制病毒潜伏期，并检查体内 BART 表达上调的潜在机制。 2、我们将利用脐带血人源化小鼠模型来探讨EBNA3A蛋白在体内的作用。目标 3，我们将探索阻断必需药物（CK4/6 和 EZH2 抑制剂）的治疗潜力该项目主要与项目 3 和 4 相互作用，并使用核心。用于免疫组织化学、生物信息学和 EBV 突变基因组的生成。研究应提供对严格 EBV 感染导致的机制的关键见解体内淋巴瘤，并可能确定治疗 EBV 诱导的淋巴瘤的新治疗方法。