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：
8196893
负责人：
ELLIOTT D KIEFF
金额：
$ 35.93万
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/8196893
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 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

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.

爱泼斯坦 - 巴尔病毒（EBV）潜在感染几乎引起所有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母细胞增殖，艾滋病患者和其他免疫损害的人中的淋巴增生性疾病国家和EBV相关的淋巴瘤，霍奇金氏病和鼻型癌。 EBV基因组在所有潜在感染的细胞中都持续存在为非集成的细胞多拷贝沿着。 EBV插图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与特定位点结合 EBV沿着ebv sistome并增强了围激曲线的初始复制，转录和持久性。由于ebna1对于在所有分裂中的EBV; 恶性细胞，该提案的核心目的是确定可以抑制EBNA1介导的偶发持续性。为了实现这一目标，我们建议：（1）进行筛选以识别中断EBNA1-ORIP依赖的化合物体内的沿着体内的转录和持久性，中断EBNA1的化合物在体外二聚化并与同源DNA结合，并与EBNA1结合的化合物在计算机中。（2）确定已鉴定化合物的生物学和生化作用在B淋巴细胞中依赖于EBNA1-ORIP依赖性偶发性转录和持久性上， EBV转化的淋巴母细胞（LCL）生长，在LCL诱导的淋巴瘤上裸鼠中的NPC肿瘤。（3）确定生物活性化合物效应的位点 EBNA1结合，使用生化，生物物理和结构方法。使用此知识最有效地进行结构活动修改以改进复合活动和特异性。（4）使用反向遗传学来识别关键 EBNA1 DBD中的残留物可以提高筛选灵敏度并告知硅口袋选择，化合物修饰和复合交互分析。