The long term goal of this research has been to determine the role of EBV in the etiology of nasopharyngeal carcinoma (NPC), an epithelial malignancy that develops with high incidence in Southern China, Northern Africa, and among Alaskan Inuits. We have previously identified the viral genes that are expressed in NPC and assessed the molecular properties of these proteins on cellular signaling and growth regulation. Our studies have shown that latent membrane protein 1 (LMP1) is expressed in NPC and has potent effects on cellular transcription, multiple cell signaling pathways, cell growth regulation, and biologic properties. This application is based on the hypothesis that specific genes are induced by the distinct transcriptional complexes induced by LMP1 and that the pleiotropic effects of LMP1 on signaling are mediated by unique complexes that form within plasma membrane domains and cytoplasmic vesicles. We suggest that LMP1 modulates the constituents of lipid raft domains to form exosomes that affect the growth of neighboring cells via a paracrine mechanism.
Our specific aims are: 1.) Identify the epithelial cellular targets that are regulated by LMP1-CTAR1 through the p50/p50-Bcl-3 transcriptional complex and activated STAT3 using chromatin immunoprecipitation (ChIP) analysis and ChIP-seq technology. 2.) Determine the composition of signaling complexes induced by LMP1 and mutants of LMP1 that have distinct functional properties using immunofluorescence, cell fractionation, immunoprecipitation, 2-D fluorescent differential gel electrophoresis, and mass spectrometry. 3.) Characterize exosomes that are produced by cells expressing LMP1. Identify their composition using immunoselection and mass spectrometry, and assess the biologic effects in cells exposed to the induced exosomes.

Public Health Relevance

Epstein Barr virus is considered a cause of multiple cancers and is consistently detected in nasopharyngeal carcinoma (NPC), a malignancy of epithelial cells that occurs at very high incidence in some populations. We have shown that the viral proteins of EBV can change the growth regulation of cells in culture and in mouse models through major effects on cellular gene expression and growth control and that these same changes occur in NPC. This grant will continue to study latent membrane protein 1 (LMP1), a viral gene that regulates cellular expression through the effects on transcription factor complexes and activates multiple cellular signaling pathways to activate cell growth. This application will identify the individual genes that are regulated by specific transcription complexes and determine the mechanisms through which LMP1 activates distinct signaling pathways.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01CA032979-30
Application #
8676433
Study Section
Virology - A Study Section (VIRA)
Program Officer
Daschner, Phillip J
Project Start
Project End
Budget Start
Budget End
Support Year
30
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Marquitz, Aron R; Mathur, Anuja; Chugh, Pauline E et al. (2014) Expression profile of microRNAs in Epstein-Barr virus-infected AGS gastric carcinoma cells. J Virol 88:1389-93
Fotheringham, Julie A; Raab-Traub, Nancy (2013) Epstein-Barr virus latent membrane protein 2 effects on epithelial acinus development reveal distinct requirements for the PY and YEEA motifs. J Virol 87:13803-15
Meckes Jr, David G; Menaker, Nathan F; Raab-Traub, Nancy (2013) Epstein-Barr virus LMP1 modulates lipid raft microdomains and the vimentin cytoskeleton for signal transduction and transformation. J Virol 87:1301-11
Meckes Jr, David G; Gunawardena, Harsha P; Dekroon, Robert M et al. (2013) Modulation of B-cell exosome proteins by gamma herpesvirus infection. Proc Natl Acad Sci U S A 110:E2925-33
Shair, Kathy H Y; Bendt, Katharine M; Edwards, Rachel H et al. (2012) Epstein-Barr virus-encoded latent membrane protein 1 (LMP1) and LMP2A function cooperatively to promote carcinoma development in a mouse carcinogenesis model. J Virol 86:5352-65
Marquitz, Aron R; Mathur, Anuja; Nam, Cyd Stacy et al. (2011) The Epstein-Barr Virus BART microRNAs target the pro-apoptotic protein Bim. Virology 412:392-400
Kung, Che-Pei; Meckes Jr, David G; Raab-Traub, Nancy (2011) Epstein-Barr virus LMP1 activates EGFR, STAT3, and ERK through effects on PKCdelta. J Virol 85:4399-408
Meckes Jr, David G; Raab-Traub, Nancy (2011) Microvesicles and viral infection. J Virol 85:12844-54
Kung, Che-Pei; Raab-Traub, Nancy (2010) Epstein-Barr virus latent membrane protein 1 modulates distinctive NF- kappaB pathways through C-terminus-activating region 1 to regulate epidermal growth factor receptor expression. J Virol 84:6605-14
Meckes Jr, David G; Shair, Kathy H Y; Marquitz, Aron R et al. (2010) Human tumor virus utilizes exosomes for intercellular communication. Proc Natl Acad Sci U S A 107:20370-5

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