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：
8400899
负责人：
ELLIOTT D KIEFF
金额：
$ 33.77万
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/8400899
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 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 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 screening small molecule tumor

项目摘要

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 proposal 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.

EB 病毒 (EBV) 潜伏感染导致几乎所有 EBV 相关发病和死亡率，包括传染性单核细胞增多症早期的淋巴母细胞增殖，艾滋病和其他免疫功能低下患者的淋巴增殖性疾病州，以及 EBV 相关淋巴瘤、霍奇金病和鼻咽癌癌。 EBV 基因组作为非整合体持续存在于所有潜伏感染的细胞中。多拷贝附加体。 EBV 附加体在分裂细胞中的持久性取决于 EBV 编码核抗原 1 蛋白 (EBNA1)。 EBNA1 结合到特定位点 EBV 附加体并增强附加体的初始复制、转录和持久性。由于 EBNA1 对于 EBV 附加体在所有分裂和分裂过程中的持续存在至关重要。恶性细胞，该提案的中心目标是识别能够抑制 EBNA1 介导的附加体持久性。为了实现这一目标，我们建议： (1) 进行筛选以识别中断 EBNA1-oriP 依赖性的化合物附加体转录和体内持久性，中断 EBNA1 的化合物体外二聚化和与同源 DNA 的结合，以及与 EBNA1 结合的化合物在计算机中。 (2) 鉴定已鉴定化合物的生物学和生化作用 EBNA1-oriP 依赖的附加体转录和 B 淋巴细胞中的持久性 EBV 转化的类淋巴母细胞 (LCL) 生长以及 LCL 诱导的淋巴瘤和裸鼠鼻咽癌肿瘤。 (3) 确定生物活性化合物作用的位点使用生物化学、生物物理和结构方法进行 EBNA1 结合。用这个最有效地进行结构活动修改以改进的知识化合物的活性和特异性。 (4)利用反向遗传学来识别关键的 EBNA1 DBD 中的残基可以提高筛选灵敏度并提供计算机信息口袋选择、化合物修饰和化合物相互作用分析。