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.

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.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA131354-05
Application #
8400899
Study Section
Drug Discovery and Mechanisms of Antimicrobial Resistance Study Section (DDR)
Program Officer
Daschner, Phillip J
Project Start
2008-12-10
Project End
2013-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
5
Fiscal Year
2013
Total Cost
$337,720
Indirect Cost
$148,521
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Jiang, Sizun; Willox, Bradford; Zhou, Hufeng et al. (2014) Epstein-Barr virus nuclear antigen 3C binds to BATF/IRF4 or SPI1/IRF4 composite sites and recruits Sin3A to repress CDKN2A. Proc Natl Acad Sci U S A 111:421-6
Chen, Ya-Lin; Liu, Cheng-Der; Cheng, Chi-Ping et al. (2014) Nucleolin is important for Epstein-Barr virus nuclear antigen 1-mediated episome binding, maintenance, and transcription. Proc Natl Acad Sci U S A 111:243-8
Lee, Eun Kyung; Kim, Sun Young; Noh, Ka-Won et al. (2014) Small molecule inhibition of Epstein-Barr virus nuclear antigen-1 DNA binding activity interferes with replication and persistence of the viral genome. Antiviral Res 104:73-83
Portal, Daniel; Zhou, Hufeng; Zhao, Bo et al. (2013) Epstein-Barr virus nuclear antigen leader protein localizes to promoters and enhancers with cell transcription factors and EBNA2. Proc Natl Acad Sci U S A 110:18537-42
Liu, Cheng-Der; Cheng, Chi-Ping; Fang, Jia-Shih et al. (2013) Modulation of Epstein-Barr virus nuclear antigen 2-dependent transcription by protein arginine methyltransferase 5. Biochem Biophys Res Commun 430:1097-102
Kim, Sun Young; Song, Kyung-A; Kieff, Elliott et al. (2012) Small molecule and peptide-mediated inhibition of Epstein-Barr virus nuclear antigen 1 dimerization. Biochem Biophys Res Commun 424:251-6
Liu, Cheng-Der; Chen, Ya-Lin; Min, Yi-Li et al. (2012) The nuclear chaperone nucleophosmin escorts an Epstein-Barr Virus nuclear antigen to establish transcriptional cascades for latent infection in human B cells. PLoS Pathog 8:e1003084
Rozenblatt-Rosen, Orit; Deo, Rahul C; Padi, Megha et al. (2012) Interpreting cancer genomes using systematic host network perturbations by tumour virus proteins. Nature 487:491-5
Xing, Li; Kieff, Elliott (2011) cis-Acting effects on RNA processing and Drosha cleavage prevent Epstein-Barr virus latency III BHRF1 expression. J Virol 85:8929-39
Zhao, Bo; Zou, James; Wang, Hongfang et al. (2011) Epstein-Barr virus exploits intrinsic B-lymphocyte transcription programs to achieve immortal cell growth. Proc Natl Acad Sci U S A 108:14902-7

Showing the most recent 10 out of 13 publications