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：
7583461
负责人：
ELLIOTT D KIEFF
金额：
$ 36.59万
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/7583461
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 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 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 Nuclear Proteins 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 public health relevance 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）：Epstein-Barr病毒（EBV）潜在感染几乎引起EBV相关的发病率和死亡率，包括感染性单核细胞增多症早期的淋巴细胞增殖，淋巴细胞增生，淋巴细胞增生性疾病，具有艾滋病患者和其他免疫损害状态，以及其他免疫损害状态，以及与EBV相关的淋巴细胞，Hodggkin，Hodggkin的疾病疾病和鼻癌癌。 EBV基因组在所有潜在感染的细胞中都持续为非集成的多拷贝曲线组。 EBV发作在分裂细胞中的持久性取决于EBV编码的核抗原1蛋白（EBNA1）。 EBNA1与EBV沿着EBV的特定位点结合，并增强了沿着沿着围激的初始复制，转录和持久性。由于EBNA1对于在所有分裂和恶性细胞中EBV发作的持久性至关重要，因此本应用的核心目的是识别可以抑制EBNA1介导的偶发持续性的化合物。为了实现这一目标，我们建议：（1）进行筛选以识别中断EBNA1-ORIP依赖性的偶发子转录和体内持久性的化合物，会中断EBNA1二聚体并结合体外的Cognate DNA的化合物，并与Cognate DNA结合，并与EBNA1在EBNA1中结合的化合物。硅。（2）确定鉴定的化合物对B淋巴细胞中EBNA1-ORIP依赖性偶发子转录的生物学和生化作用，对EBV转化的淋巴母细胞（LCL）生长以及LCL诱导的Nude小鼠的淋巴瘤和NPC肿瘤对EBV转化的淋巴母细胞（LCL）生长。（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效应的位点。使用这些知识最有效地进行结构活动修改，以改善复合活动和特异性。（4）使用反向遗传学来识别EBNA1 DBD中的关键残基，这些残基可以提高筛选敏感性并告知硅袋选择，复合修饰和复合相互作用分析。公共卫生相关性：Epstein-Barr病毒（EBV）积极引起感染或其他免疫损害的人，伯基特淋巴瘤，其他淋巴瘤，霍奇金病，几乎所有鼻咽癌的艾滋病毒中的恶性淋巴增生性疾病。由于EBV编码的核抗原1（EBNA1）蛋白对于这些恶性细胞中EBV基因组的持久性至关重要，因此我们提出了识别和改善抗EBNA1化合物并防止EBV基因组持久性的实验。这些化合物可能预防或阻止EBV相关的肿瘤细胞生长并导致肿瘤细胞死亡。