Inhibitors of Epstein-Barr Virus Nuclear Protein 1 Mediated Latent Infection

EB 病毒核蛋白 1 抑制剂介导的潜伏感染

• 批准号: 7988583
• 负责人: ELLIOTT D KIEFF
• 金额: $ 35.84万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-10 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988583
• 关键词: AIDS-Related Lymphoma; Acquired Immunodeficiency Syndrome; Antisense Oligonucleotides; Arginine; B-Lymphocytes; Binding; Biochemical; Biological; Biological Assay; Burkitt Lymphoma; Carcinoma; Cell Death; Cell Nucleus; Cell Survival; Cells; Centers for Disease Control and Prevention (U.S.); Chromosomes; Computer Simulation; DNA; DNA Binding; DNA Binding Domain; DNA biosynthesis; DNA-Protein Interaction; Dimerization; Disease; Dominant-Negative Mutation; EBV-associated disease; EBV-encoded nuclear antigen 1; Elements; Episome; Epithelial Cells; Gene Expression; Genetic Transcription; Genome; Glycine; HIV; Health; Hodgkin Disease; Human; Human Herpesvirus 4; Immune; In Vitro; Infectious Mononucleosis; Knowledge; Large-Cell Immunoblastic Lymphoma; Lead; Ligands; Lymphocyte; Lymphoma; Lymphoproliferative Disorders; Malignant - descriptor; Mediating; Modification; Morbidity - disease rate; Mutation; Nasopharynx Carcinoma; Nuclear; Nuclear Antigens; Nuclear Pore Complex; Nuclear Protein; Nude Mice; Oropharyngeal; Plasmids; Precursor B-Lymphoblast; Prevention; Protein Binding; Protein Inhibition; Proteins; RNA Interference; Reporting; Screening procedure; Site; Specificity; Structure; Transplant Recipients; Viral Genes; base; cancer cell; cell growth; improved; in vivo; inhibitor/antagonist; latent infection; lymphoblast; mortality; neoplastic cell; positional cloning; prevent; protein function; research study; small molecule; tumor

DESCRIPTION (provided by applicant): Epstein-Barr Virus (EBV) latent infections cause almost all EBV associated morbidity and mortality including lymphoblast proliferation early in Infectious Mononucleosis, Lymphoproliferative Diseases in people with AIDS and other immune compromised states, and EBV associated Lymphomas, Hodgkin's Disease, and Nasophryngeal Carcinoma. The EBV genome persists in all latently infected cells as a non-integrated multi-copy episome. The persistence of EBV episomes in dividing cells is dependent on the EBV encoded nuclear antigen 1 protein (EBNA1). EBNA1 binds to a specific site in the EBV episome and enhances episome initial replication, transcription, and persistence. Since EBNA1 is essential for the persistence of EBV episomes in all dividing and malignant cells, the central objective of this application is to identify compounds that can inhibit EBNA1 mediated episome persistence. To achieve that objective, we propose to: (1) Undertake screens to identify compounds that interrupt EBNA1-oriP dependent episome transcription and persistence in vivo, compounds that interrupt EBNA1 dimerization and binding to cognate DNA in vitro, and compounds that bind to EBNA1 in silico. (2) Identify the biological and biochemical effects of the identified compounds on EBNA1-oriP dependent episome transcription and persistence in B lymphoblasts, on EBV transformed lymphoblastoid cell (LCL) growth, and on LCL induced Lymphoma and NPC tumors in nude mice. (3) Determine the sites of bioactive compound effects in EBNA1 binding, using biochemical, biophysical, and structural approaches. Use this knowledge to most effectively undertake structure activity modifications to improve compound activity and specificity. (4) Use reverse genetics to identify the critical residues in EBNA1 DBD that can improve screening sensitivity and inform in silico pocket selection, compound modification, and compound interaction analyses. PUBLIC HEALTH RELEVANCE: Epstein-Barr Virus (EBV) actively causes malignant lymphoproliferative diseases in HIV infected or otherwise immune compromised people, Burkitt Lymphoma, other Lymphomas, Hodgkin's Disease, and almost all Nasopharyngeal Carcinomas. Since the EBV-encoded nuclear antigen 1 (EBNA1) protein is essential for the persistence of the EBV genome in these malignant cells, we propose experiments that identify and improve anti-EBNA1 compounds and prevent EBV genome persistence. These compounds are likely to prevent or halt EBV associated tumor cell growth and cause tumor cell death.

描述(申请人提供)：EB病毒(EBV)潜伏感染导致几乎所有与EBV相关的发病率和死亡率，包括传染性单核细胞增多症早期的淋巴母细胞增殖，艾滋病患者的淋巴增殖性疾病和其他免疫受损状态，以及EBV相关性淋巴瘤、霍奇金氏病和鼻咽癌。EBV基因组在所有潜伏感染的细胞中作为一个非整合的多拷贝Episome持续存在。EB病毒上皮体在细胞分裂中的持久性依赖于EBV编码的核抗原1蛋白(EBNA1)。EBNA1与EBV Episome中的特定位点结合，增强Episome的初始复制、转录和持久性。由于EBNA1对EBV episome在所有分裂和恶性细胞中的持久性是必不可少的，本应用的中心目标是确定能够抑制EBNA1介导的Episome持久性的化合物。为了实现这一目标，我们建议：(1)进行筛选，以确定在体内中断EBNA1-ORIP依赖的Episome转录和持久性的化合物，在体外中断EBNA1二聚并与同源DNA结合的化合物，以及在电子计算机中与EBNA1结合的化合物。(2)鉴定化合物对EBNA1-ORIP依赖的Episome转录和在B淋巴母细胞中的持久性、对EBV转化的淋巴母细胞(LCL)生长以及对LCL诱发的淋巴瘤和鼻咽癌裸鼠体内的生物学和生化影响。(3)利用生化、生物物理和结构分析等方法，确定EBNA1结合中生物活性化合物的作用部位。利用这些知识最有效地进行结构活性修饰，以提高化合物的活性和特异性。(4)利用反向遗传学来识别EBNA1 DBD中的关键残基，以提高筛选的敏感性，并在电子筛选、化合物修饰和化合物相互作用分析中提供信息。公共卫生相关性：爱泼斯坦-巴尔病毒(EBV)在HIV感染者或其他免疫受损的人、Burkitt淋巴瘤、其他淋巴瘤、霍奇金氏病和几乎所有鼻咽癌中积极引起恶性淋巴增生性疾病。由于EBV编码的核抗原1(EBNA1)蛋白对于EBV基因组在这些恶性肿瘤细胞中的持续存在是必不可少的，我们建议进行实验，以识别和改进抗EBNA1化合物，并防止EBV基因组持续存在。这些化合物可能会阻止或阻止EBV相关肿瘤细胞的生长，并导致肿瘤细胞死亡。