Genetic and Proteomic Analysis of Epstein-Barr Virus LMP1 Activation of NF-kB

EB 病毒 LMP1 激活 NF-kB 的遗传和蛋白质组学分析

• 批准号: 8068346
• 负责人: Benjamin Elison Gewurz
• 金额: $ 17.99万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8068346
• 关键词: Adverse effects; Affect; Apoptosis; Autoimmune Process; Autoimmunity; B-Lymphocytes; Biochemical; Biochemical Genetics; Biological Assay; Cancerous; Cell Death; Cell Proliferation; Cell Survival; Cells; Chemicals; Clinical; Communicable Diseases; Complex; Cytoplasmic Tail; Deubiquitinating Enzyme; Development; Disease; Drug Delivery Systems; Enzyme Inhibition; Enzymes; Epithelial; Epithelial Cells; Epstein-Barr Virus Infections; Epstein-Barr virus LMP-1 protein; Genetic; Genetic Transcription; Grant; Growth; HIV; Hodgkin Disease; Host Defense; Human; Human Genome; Human Herpesvirus 4; Hypersensitivity; I Kappa B-Alpha; Immune; Immunocompromised Host; Immunologic Receptors; Immunologic Surveillance; Immunology; Inflammation; Knowledge; Ligands; Lymphoma; Lymphoproliferative Disorders; MAP3K7 gene; MAP3K7IP1 gene; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane Proteins; Mitogen-Activated Protein Kinases; NF-kappa B; Nasopharynx Carcinoma; Nuclear; Oncogene Proteins; Oncogenes; Oncogenic; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphotransferases; Precursor B-Lymphoblast; Prevention; Proteins; Proteomics; RNA Interference; Reporter; Research; Research Personnel; Role; Signal Transduction; Site; Small Interfering RNA; Systems Biology; TRAF6 gene; Therapeutic; Throat Cancer; Toxic effect; Training; Transplantation; Virus Diseases; Work; base; career; cell growth; cell transformation; chemotherapy; design; functional group; gammaherpesvirus; genetic analysis; genome wide association study; genome-wide; inhibitor/antagonist; interest; loss of function; lymphoblast; microbial; neoplastic; novel; persistent EBV infection; public health relevance; scaffold; small molecule; therapeutic target; trafficking; transcription factor; tumor; tumorigenesis; ubiquitin ligase; ubiquitin-specific protease; virology

DESCRIPTION (provided by applicant): The Epstein Barr Virus (EBV) is an oncogenic gamma-herpesvirus that is associated with Hodgkin disease and anaplasmic nasopharyngeal carcinoma. EBV infection is particularly hazardous with advanced HIV disease or transplant, where EBV-encoded proteins drive aberrant cell growth in the absence of immune surveillance. The principal EBV oncogene, Latent Membrane Protein 1 (LMP1), promotes cell survival and proliferation by mimicking activated immune receptors. Through incompletely defined pathways, LMP1 potently stimulates Nuclear Factor Kappa B (NF-kB), transcription factors that control inflammation, cell survival and growth. EBV- transformed cells rely on constitutive NF-kB activation, and rapidly undergo apoptosis upon NF-kB blockade. Side-effects preclude the clinical use of currently available NF-kB inhibitors, though LMP1-selective drug targets may afford substantially less toxicity. It is therefore important to define how LMP1 activates NF-kB. I have carried out a human genome-wide siRNA screen for cellular modulators of LMP1 canonical NF-kB activation. Hits that either suppress or enhance LMP1 activation of NF-kB have been validated with secondary screens. The screens have implicated numerous proteins in LMP1 function, including novel factors not previously associated with NF-kB. I will carry out hypothesis-based and larger-scale secondary assays to identify critical missing components of LMP1 NF-kB activation in both epithelial cells and B lymphoblasts. I will pursue detailed biochemical analysis of several targets of particular interest, including potentially druggable enzymes and functionally clustered hits. Secondary screens will further stratify hits into functional groups based on whether they affect LMP1 expression, subcellular trafficking, and where they function within the LMP1/NF-kB pathway. Cellular factors uniquely employed by LMP1, but not by immune receptor pathways, may serve as important therapeutic targets for treatment of EBV-driven malignancies. Likewise, these studies may reveal important general mechanisms of NF-kB activation, with implications for allergy, autoimmunity, and host-defense. PUBLIC HEALTH RELEVANCE: Persistent Epstein Barr virus infection is an important cause of certain lymphoma and throat cancers. This project will better define how EBV subverts cellular machinery to drive cancerous growth of infected cells. Ultimately, it is hoped that knowledge gained from these studies may enable the development of chemotherapies that specifically block EBV function.

描述（由申请人提供）：EB病毒（EBV）是一种致癌性γ-疱疹病毒，与霍奇金病和无形体鼻咽癌相关。 EBV 感染对于晚期 HIV 疾病或移植尤其危险，其中 EBV 编码的蛋白质在缺乏免疫监视的情况下会驱动异常细胞生长。 EBV 的主要癌基因潜膜蛋白 1 (LMP1) 通过模仿激活的免疫受体来促进细胞存活和增殖。通过不完全定义的途径，LMP1 有效刺激核因子 Kappa B (NF-kB)，这是控制炎症、细胞存活和生长的转录因子。 EBV 转化细胞依赖于 NF-kB 的组成型激活，并在 NF-kB 阻断后迅速发生凋亡。尽管 LMP1 选择性药物靶点可能会显着降低毒性，但副作用阻碍了目前可用的 NF-kB 抑制剂的临床使用。因此，定义 LMP1 如何激活 NF-kB 非常重要。我对 LMP1 典型 NF-kB 激活的细胞调节剂进行了人类全基因组 siRNA 筛选。抑制或增强 NF-kB 的 LMP1 激活的命中已通过二次筛选得到验证。这些筛选表明许多蛋白质与 LMP1 功能有关，包括以前与 NF-kB 不相关的新因子。我将进行基于假设的大规模二次测定，以确定上皮细胞和 B 淋巴母细胞中 LMP1 NF-kB 激活的关键缺失成分。我将对几个特别感兴趣的目标进行详细的生化分析，包括潜在的可药物酶和功能性聚集的命中。二次筛选将根据命中是否影响 LMP1 表达、亚细胞运输以及它们在 LMP1/NF-kB 通路中的作用位置，进一步将命中分为功能组。 LMP1 独有的细胞因子（而非免疫受体途径）可能作为治疗 EBV 驱动的恶性肿瘤的重要治疗靶点。同样，这些研究可能揭示 NF-kB 激活的重要一般机制，对过敏、自身免疫和宿主防御具有影响。 公共卫生相关性：持续的 Epstein Barr 病毒感染是某些淋巴瘤和喉癌的重要原因。该项目将更好地定义 EBV 如何颠覆细胞机制以驱动受感染细胞的癌性生长。最终，希望从这些研究中获得的知识能够开发出特异性阻断 EBV 功能的化疗药物。