The transforming human gammaherpesviruses EBV and KSHV establish stable latent infections in B cells, providing a lifelong reservoir of virus tha can contribute to the development of malignant disease. Thus, defining the mechanisms that govern long-term latency in B cells is critical for designing rational strategies to prevent diseas. In vivo studies of gammaherpesviruses in humans have been severely limited by the difficulties of working in the natural host. Murine gammaherpesvirus 68 (MHV68) is related to EBV and KSHV and causes lymphomas and lymphoproliferative disease in mice, providing a readily manipulable small animal model for mechanistic studies of the virus/host relationship in vivo. Like EBV and KSHV, MHV68 expresses a defined set of miRNAs whose functions during infection are largely unknown. As these virus-encoded miRNAs are abundantly expressed in vivo in B cells during long-term latency and in hyperplastic B cell lesions during lymphoproliferative disease, we hypothesize that these viral miRNAs play key roles in latency and pathogenesis. To test this hypothesis, we will 1) Determine the functional role of individual MHV68 miRNAs in long-term latency and lymphoproliferative disease; 2) Identify specific virus and host targets of the key miRNAs using cutting-edge CLIP and deep sequencing approaches coupled with in vitro and in vivo validation; 3) Define the in vivo biological relevance of target genes and determine their roles in B cell maturation and survival. We anticipate that there will be significant overlap between the mRNA targets and cellular pathways regulated by MHV68, KSHV, and EBV miRNAs. The systematic in vivo analyses of MHV68 miRNA mutants, in conjunction with HITS-CLIP technology and in vivo validation of miRNA targets, provides an extremely powerful means to determine the molecular mechanism by which miRNAs contribute to gammaherpesvirus infection in vivo, and should allow us to define the contribution of repression of these shared targets to gammaherpesvirus latency and pathogenesis.

Public Health Relevance

Gammaherpesviruses such as EBV and KSHV establish dormant lifelong infections in humans, and have been linked to the development of numerous types of cancers. Research here will use a mouse model to examine the role that gammaherpesvirus microRNAs play in lifelong infection and induction of disease. This work has important implications for the development of drugs to prevent or treat such virus infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI108407-03
Application #
9195696
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Beisel, Christopher E
Project Start
2015-01-01
Project End
2019-12-31
Budget Start
2017-01-01
Budget End
2017-12-31
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Florida
Department
Genetics
Type
Schools of Medicine
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Van Skike, Nick D; Minkah, Nana K; Hogan, Chad H et al. (2018) Viral FGARAT ORF75A promotes early events in lytic infection and gammaherpesvirus pathogenesis in mice. PLoS Pathog 14:e1006843
Bullard, Whitney L; Flemington, Erik K; Renne, Rolf et al. (2018) Connivance, Complicity, or Collusion? The Role of Noncoding RNAs in Promoting Gammaherpesvirus Tumorigenesis. Trends Cancer 4:729-740
Chen, Hao; Bartee, Mee Yong; Yaron, Jordan R et al. (2018) Mouse Gamma Herpesvirus MHV-68 Induces Severe Gastrointestinal (GI) Dilatation in Interferon Gamma Receptor-Deficient Mice (IFN?R-/-) That Is Blocked by Interleukin-10. Viruses 10:
Ungerleider, Nathan; Concha, Monica; Lin, Zhen et al. (2018) The Epstein Barr virus circRNAome. PLoS Pathog 14:e1007206
Feldman, Emily R; Kara, Mehmet; Oko, Lauren M et al. (2016) A Gammaherpesvirus Noncoding RNA Is Essential for Hematogenous Dissemination and Establishment of Peripheral Latency. mSphere 1:
Feldman, Emily R; Tibbetts, Scott A (2015) Emerging Roles of Herpesvirus microRNAs During In Vivo Infection and Pathogenesis. Curr Pathobiol Rep 3:209-217