Epstein-Barr virus (EBV) causes or is associated with a wide range of diseases varying in severity from infectious mononucleosis to life-threatening lymphoproliferative diseases in immunocompromised patients. The consistent presence of EBV in particular tumor types, such as AIDS-associated CNS and other B-cell lymphomas, offers the potential for the development of highly specific, virus-targeted therapies. This proposal combines basic studies of a novel potential target, EBV BGLF4-encoded protein kinase (EBV PK), with applied studies, wherein the kinase will be tested as a """"""""suicide"""""""" gene for treatment of EBV-positive and EBV negative tumors (in conjunction with ganciclovir treatment) or as a """"""""therapeutic"""""""" gene for treatment of EBV driven lymphomas.
In Aim 1, we study the role of EBV PK in viral infection by knocking out its expression using RNAi. We then analyze the course of viral lytic infection at the RNA and protein levels, starting with known EBV PK targets, such as viral DMA polymerase processivity factor, encoded by the EBV BMRF1 gene, and extending the research to newly discovered targets.
In Aim 2, we investigate the phosphorylation of EBV EBNA2 protein, the master regulator of the EBV type 3 latency program characteristic of EBV lymphoproliferative diseases, and a newly discovered EBV PK target. Based on this finding, we also propose a novel potential therapeutic approach for treating EBV-driven lymphoproliferations, that is, by deregulation of type 3 EBV latency.
In Aim 3, we develop ways to use EBV kinases, both PK and thymidine kinase (TK), to treat EBV-positive and EBV-negative malignancies. We study, first in cell culture and then in animal models of human disease, the efficacy of the kinases in converting ganciclovir to its cytotoxic form, and the ability of combinations of chemotherapeutic agents that trigger viral reactivation and ganciclovir to inhibit the growth of lymphomas containing wild type EBV or mutants lacking either kinase. Hence this research will reveal the role of EBV PK in viral infection and ultimately lead to new approaches for treatment of important EBV-related malignancies, based on new concepts rooted in fundamentals of the virology of EBV.
|Ning, S; Pagano, J S; Barber, G N (2011) IRF7: activation, regulation, modification and function. Genes Immun 12:399-414|
|Bentz, Gretchen L; Whitehurst, Christopher B; Pagano, Joseph S (2011) Epstein-Barr virus latent membrane protein 1 (LMP1) C-terminal-activating region 3 contributes to LMP1-mediated cellular migration via its interaction with Ubc9. J Virol 85:10144-53|
|Bentz, Gretchen L; Liu, Renshui; Hahn, Angela M et al. (2010) Epstein-Barr virus BRLF1 inhibits transcription of IRF3 and IRF7 and suppresses induction of interferon-beta. Virology 402:121-8|
|Whitehurst, Christopher B; Ning, Shunbin; Bentz, Gretchen L et al. (2009) The Epstein-Barr virus (EBV) deubiquitinating enzyme BPLF1 reduces EBV ribonucleotide reductase activity. J Virol 83:4345-53|
|Wang, Fu-Zhang; Roy, Debasmita; Gershburg, Edward et al. (2009) Maribavir inhibits epstein-barr virus transcription in addition to viral DNA replication. J Virol 83:12108-17|
|Gershburg, Edward; Pagano, Joseph S (2008) Conserved herpesvirus protein kinases. Biochim Biophys Acta 1784:203-12|
|Gershburg, Edward; Raffa, Salvatore; Torrisi, Maria Rosaria et al. (2007) Epstein-Barr virus-encoded protein kinase (BGLF4) is involved in production of infectious virus. J Virol 81:5407-12|
|Yue, Wei; Gershburg, Edward; Pagano, Joseph S (2005) Hyperphosphorylation of EBNA2 by Epstein-Barr virus protein kinase suppresses transactivation of the LMP1 promoter. J Virol 79:5880-5|
|Gershburg, E; Marschall, M; Hong, K et al. (2004) Expression and localization of the Epstein-Barr virus-encoded protein kinase. J Virol 78:12140-6|
|Feng, Wen-hai; Cohen, Jeffrey I; Fischer, Steven et al. (2004) Reactivation of latent Epstein-Barr virus by methotrexate: a potential contributor to methotrexate-associated lymphomas. J Natl Cancer Inst 96:1691-702|
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