Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that is etiologically linked to several human cancers, including AIDS-associated B-cell lymphomas. Previously, we have reported that both EBV-encoded microRNAs (miRNAs) and EBV-induced cellular miRNAs play a critical role in promoting cellular transformation by EBV in culture. Here, we propose an integrated series of experiments designed to lead to a mechanistic understanding of how the miRNAs encoded by, or induced by, EBV infection contribute to the growth and transformation of human cells in culture and in vivo. We will dissect the mechanisms used by EBV-encoded and EBV-induced cellular miRNAs to facilitate the formation of stably immortalized lymphoblastoid cell lines (LCLs) and, more generally, examine how miRNAs contribute to lymphomagenesis and control the decision between viral latency and lytic replication. In addition, we will determine whether specific viral or cellular miRNAs promote the growth and viability of latently EBV-infected diffuse large B cell lymphoma cells (DLBCLs) and nasopharyngeal carcinoma (NPC) cells, in order to ascertain whether EBV uses common mechanisms to transform B lymphocytes and epithelial cells. While one important focus of this grant will be on a cluster of three EBV-encoded miRNAs (miR-BHRF1-1, 1-2 and 1-3) that is expressed at high levels in LCLs and in many DLBCLs, we will also address how the 22-miRNA EBV BART cluster affects the ability of EBV-transformed cells to grow in culture and to avoid immune elimination. Critically, we will integrate miRNA target identification, using a powerful cross-linking and immunoprecipitation (CLIP) technique, with phenotypic studies of miRNA:mRNA interactions to define the mechanisms of action used by these miRNAs to mediate EBV latency and pathogenesis.

Public Health Relevance

Epstein-Barr virus (EBV) is a herpesvirus that is linked to several human diseases, including AIDS- associated B-cell lymphomas and nasopharyngeal carcinomas. We have observed that microRNAs, either those encoded by EBV itself or cellular microRNAs activated upon infection, play an important role in the establishment and/or maintenance of cellular transformation induced by EBV. This grant seeks to understand the mechanisms underlying this process with a view to the development of potential new treatment modalities.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI067968-09
Application #
8849809
Study Section
AIDS-associated Opportunistic Infections and Cancer Study Section (AOIC)
Program Officer
Beisel, Christopher E
Project Start
2005-12-01
Project End
2016-05-31
Budget Start
2015-06-01
Budget End
2016-05-31
Support Year
9
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Duke University
Department
Genetics
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Poling, Brigid Chiyoko; Price, Alexander M; Luftig, Micah A et al. (2017) The Epstein-Barr virus miR-BHRF1 microRNAs regulate viral gene expression in cis. Virology 512:113-123
Poling, Brigid Chiyoko; Tsai, Kevin; Kang, Dong et al. (2017) A lentiviral vector bearing a reverse intron demonstrates superior expression of both proteins and microRNAs. RNA Biol 14:1570-1579
Gregorovic, Goran; Boulden, Elizabeth A; Bosshard, Rachel et al. (2015) Epstein-Barr Viruses (EBVs) Deficient in EBV-Encoded RNAs Have Higher Levels of Latent Membrane Protein 2 RNA Expression in Lymphoblastoid Cell Lines and Efficiently Establish Persistent Infections in Humanized Mice. J Virol 89:11711-4
Kang, Dong; Skalsky, Rebecca L; Cullen, Bryan R (2015) EBV BART MicroRNAs Target Multiple Pro-apoptotic Cellular Genes to Promote Epithelial Cell Survival. PLoS Pathog 11:e1004979
Skalsky, Rebecca L; Cullen, Bryan R (2015) EBV Noncoding RNAs. Curr Top Microbiol Immunol 391:181-217
Kennedy, Edward M; Cullen, Bryan R (2015) Bacterial CRISPR/Cas DNA endonucleases: A revolutionary technology that could dramatically impact viral research and treatment. Virology 479-480:213-20
Parnas, Oren; Corcoran, David L; Cullen, Bryan R (2014) Analysis of the mRNA targetome of microRNAs expressed by Marek's disease virus. MBio 5:e01060-13
Flores, Omar; Kennedy, Edward M; Skalsky, Rebecca L et al. (2014) Differential RISC association of endogenous human microRNAs predicts their inhibitory potential. Nucleic Acids Res 42:4629-39
Skalsky, Rebecca L; Kang, Dong; Linnstaedt, Sarah D et al. (2014) Evolutionary conservation of primate lymphocryptovirus microRNA targets. J Virol 88:1617-35
Jin, Hyun Yong; Oda, Hiroyo; Lai, Maoyi et al. (2013) MicroRNA-17~92 plays a causative role in lymphomagenesis by coordinating multiple oncogenic pathways. EMBO J 32:2377-91

Showing the most recent 10 out of 35 publications