Epstein-Barr virus (EBV) is a human herpesvirus that infects both B lymphocytes and epithelial cells. Depending upon cell type and differentiation state, infection leads to lytic replication with cell death or latent infection. Latent infections are associated with several epithelial- and B-cell malignancies. By identifying key cellular factors and pathways regulating EBV latency and lytic replication and drugs that affect their activities, we hope to develop lytic induction therapies for treating patients with EBV-associated cancers. The immediate- early BZLF1 and BRLF1 genes of EBV encode multifunctional proteins essential for lytic replication. While transcription from their promoters, Zp and Rp, respectively, is strongly silenced in latently infected cells, cell differentiation and factors activated by signaling pathways can induce their expression, leading to reactivation into lytic replication. We have identifled several physiologically relevant factors that play key roles in regulating this switch: ZEB1, ZEB2, p53, Oct-1, Oct-2, Smads via TGF-p signaling, and the Zp ZIIR element. We have preliminary data indicating important roles as well for HIF-1 a, Pax5, and Blimp-la and have identified 2 drugs, nufiin-3 and L-mimosine, that induce EBV reactivation via effects on some of these factors. We have developed a mouse model with a humanized immune system that can be use to examine factors affecting EBV-induced tumorigenesis and effects of drugs on growth and killing of EBV+ lymphomas. We have also identified a non-tumorigenic epithelial cell line, NOKs, in which we can study effects of differentiation state on EBV's life cycle. Here, we propose to determine: (i) how HIF-1 a, the ZEBs, PAX5, and Blimp-la play key roles in controlling EBV's latent-lytic balance in cells in culture, during differentiation of EBV+ NOKs and B cells, and in humanized mice; and (ii) whether nufiin-3 or L-mimosine together with HDAC inhibitors can efficiently reactivate EBV, leading to highly selective killing of some types of EBV+ cells lines in culture and tumors in humanized mice, especially when used in combination with ganciclovir, a nucleoside analog prodrug activated by a virus-encoded kinase. The findings obtained from these experiments will identify key cellular factors and pathways that regulate the balance between latency and lytic replication of EBV in epithelial and B cells, may identify effective lytic-inducing drugs for treating EBV-associated cancers, and may lead to a candidate vaccine for immunization against EBV by identifying an EBV mutant unable to establish viral latency. Drs. Kenney and Mertz collaborate extensively, including holding bimonthly joint research group meetings and co-authoring publications in which both labs made key contributions.

Public Health Relevance

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus associated with several types of cancers. This project is designed to identify drugs that effectively reactivate this virus out of latency into lytic replication to use for treating patients with EBV-associated cancers. This work may lead to a candidate vaccine for immunization against EBV.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA022443-40
Application #
9265004
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
Budget Start
2017-05-01
Budget End
2018-04-30
Support Year
40
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Zumwalde, Nicholas A; Sharma, Akshat; Xu, Xuequn et al. (2017) Adoptively transferred V?9V?2 T cells show potent antitumor effects in a preclinical B cell lymphomagenesis model. JCI Insight 2:
Ma, Shi-Dong; Tsai, Ming-Han; Romero-Masters, James C et al. (2017) Latent Membrane Protein 1 (LMP1) and LMP2A Collaborate To Promote Epstein-Barr Virus-Induced B Cell Lymphomas in a Cord Blood-Humanized Mouse Model but Are Not Essential. J Virol 91:
Bilger, Andrea; Plowshay, Julie; Ma, Shidong et al. (2017) Leflunomide/teriflunomide inhibit Epstein-Barr virus (EBV)- induced lymphoproliferative disease and lytic viral replication. Oncotarget 8:44266-44280
Yang, Ya-Chun; Liem, Amy; Lambert, Paul F et al. (2017) Dissecting the regulation of EBV's BART miRNAs in carcinomas. Virology 505:148-154
Iwahori, Satoko; UmaƱa, Angie C; VanDeusen, Halena R et al. (2017) Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins. J Biol Chem 292:6583-6599
Wille, Coral K; Li, Yangguang; Rui, Lixin et al. (2017) Restricted TET2 Expression in Germinal Center Type B Cells Promotes Stringent Epstein-Barr Virus Latency. J Virol 91:
Chandra, Janin; Kuo, Paula T Y; Hahn, Anne M et al. (2017) Batf3 selectively determines acquisition of CD8+ dendritic cell phenotype and function. Immunol Cell Biol 95:215-223
Pocock, Ginger M; Zimdars, Laraine L; Yuan, Ming et al. (2017) Diverse activities of viral cis-acting RNA regulatory elements revealed using multicolor, long-term, single-cell imaging. Mol Biol Cell 28:476-487
Nowak, Karolin; Linzner, Daniela; Thrasher, Adrian J et al. (2017) Absence of ?-Chain in Keratinocytes Alters Chemokine Secretion, Resulting in Reduced Immune Cell Recruitment. J Invest Dermatol 137:2120-2130
Uberoi, Aayushi; Lambert, Paul F (2017) Rodent Papillomaviruses. Viruses 9:

Showing the most recent 10 out of 420 publications