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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：EB 病毒 (EBV) 潜伏感染导致几乎所有 EBV 相关的发病和死亡，包括传染性单核细胞增多症早期的淋巴母细胞增殖、艾滋病患者和其他免疫受损状态患者的淋巴增殖性疾病，以及 EBV 相关的淋巴瘤、霍奇金氏病疾病和鼻咽癌。 EBV 基因组作为非整合的多拷贝附加体存在于所有潜伏感染的细胞中。 EBV 附加体在分裂细胞中的持续存在取决于 EBV 编码的核抗原 1 蛋白 (EBNA1)。 EBNA1 与 EBV 附加体中的特定位点结合，增强附加体的初始复制、转录和持久性。由于 EBNA1 对于 EBV 附加体在所有分裂细胞和恶性细胞中的持久性至关重要，因此本申请的中心目标是鉴定可以抑制 EBNA1 介导的附加体持久性的化合物。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 依赖性附加体转录和持久性的化合物，在体外中断 EBNA1 二聚化和与同源 DNA 结合的化合物，以及在体外与 EBNA1 结合的化合物。硅片。 (2) 确定所鉴定的化合物对 B 淋巴母细胞中 EBNA1-oriP 依赖性附加体转录和持久性、对 EBV 转化的淋巴母细胞 (LCL) 生长以及对裸鼠中 LCL 诱导的淋巴瘤和 NPC 肿瘤的生物和生化作用。 (3) 使用生化、生物物理和结构方法确定 EBNA1 结合中生物活性化合物作用的位点。利用这些知识最有效地进行结构活性修饰，以提高化合物活性和特异性。 (4) 使用反向遗传学来鉴定 EBNA1 DBD 中的关键残基，这些残基可以提高筛选灵敏度并为硅袋选择、化合物修饰和化合物相互作用分析提供信息。公共卫生相关性：EB 病毒 (EBV) 会在 HIV 感染者或其他免疫功能低下的人群中引起恶性淋巴增殖性疾病、伯基特淋巴瘤、其他淋巴瘤、霍奇金病和几乎所有鼻咽癌。由于 EBV 编码的核抗原 1 (EBNA1) 蛋白对于 EBV 基因组在这些恶性细胞中的持续存在至关重要，因此我们提出了鉴定和改进抗 EBNA1 化合物并防止 EBV 基因组持续存在的实验。这些化合物可能会阻止或阻止 EBV 相关肿瘤细胞生长并导致肿瘤细胞死亡。