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)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA032979-29
Application #
8464009
Study Section
Virology - A Study Section (VIRA)
Program Officer
Daschner, Phillip J
Project Start
1983-06-01
Project End
2015-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
29
Fiscal Year
2013
Total Cost
$280,058
Indirect Cost
$90,829
Name
University of North Carolina Chapel Hill
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Fotheringham, Julie A; Raab-Traub, Nancy (2015) Epstein-Barr virus latent membrane protein 2 induces autophagy to promote abnormal acinus formation. J Virol 89:6940-4
Marquitz, Aron R; Mathur, Anuja; Edwards, Rachel Hood et al. (2015) Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region. J Virol 89:11256-68
Raab-Traub, Nancy (2015) Nasopharyngeal Carcinoma: An Evolving Role for the Epstein-Barr Virus. Curr Top Microbiol Immunol 390:339-63
Tworkoski, Kathryn; Raab-Traub, Nancy (2015) LMP1 promotes expression of insulin-like growth factor 1 (IGF1) to selectively activate IGF1 receptor and drive cell proliferation. J Virol 89:2590-602
Edwards, Rachel Hood; Marquitz, Aron R; Raab-Traub, Nancy (2015) Changes in expression induced by Epstein-Barr Virus LMP1-CTAR1: potential role of bcl3. MBio 6:
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; 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
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
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

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